The Ministry of Health, Labour and Welfare Ministerial Notiˆcation No. 461 In accordance with the provisions of Article 41, Paragraph 1 of the Pharmaceutical AŠairs Law (Law No. 145, 1960), we hereby revise a part of the Japanese Phar- macopoeia (Ministerial Notiˆcation No. 111, 2001) as follows, and the revised Japanese Pharmacopoeia shall come into eŠect on January 1, 2005, [including dele- tion from O‹cial Monographs fro Part II in The Japanese Pharmacopoeia, Four- teenth Edition of the articles of Absorbent Cotton, Puriˆed Absorbent Cotton, Sterile Absorbent Cotton, Sterile Puriˆed Absorbent Cotton and Absorbent Gauze and Sterile Absorbent Gauze (hereinafter referred to as ``sanitary materials'')]. Provi- so: With respect to the drugs which are included in the Japanese Pharmacopoeia (hereinafter referred to as ``the old Japanese Pharmacopoeia'') [limited to those included in the Japanese Pharmacopoeia whose standards are changed with this notiˆcation published (hereinafter referred to as ``the new Japanese Phar- macopoeia'')] and those which are approved as of January 1, 2005 pursuant to the provisions of Article 14, Paragraph 1 of this Law (including cases where it shall apply mutatis mutandis under Article 23 of this Law; the same hereinafter) [including the drugs designated as those exempted from approval (hereinafter referred to as ``the drugs exempted from approval'') among the drugs etc. designated by the Minister of Health, Labour and Welfare as those exempted from manufacturing or import approval pursuant to the provisions of Article 14, Paragraph 1 of the Pharmaceutical AŠairs Law (Ministerial Notiˆcation No. 104, 1994), the standards established in the old Japanese Pharmacopoeia (limited to the standards for the relevant drugs) shall be recognized, up to June 30, 2006, as the standards established in the new Japanese Pharmacopoeia. With respect the drugs which are included in the new Japanese Pharmacopoeia (excluding those which are included in the old Japanese Phar- macopoeia) and those which are approved as of January 1, 2005 pursuant to the provisions of Article 14, Paragraph 1 of this Law (including the drugs exempted from approval), the drugs may be treated, up to June 30, 2006, as those which are not included in the new Japanese Pharmacopoeia. Further, sanitary materials may be treated, up to September 30, 2006, under the previous regulation. Hidehisa Otsuji The Minister of Health, Labour and Welfare December 28, 2004 (The texts referred to by the term ``as follows'' are omitted here. All of them are made available for public exhibition at the Evaluation and Licensing Division, Phar- maceutical and Medical Safety Bureau, Ministry of Health, Labour and Welfare, at each Regional Bureau of Health and Welfare and at each Prefectural O‹ce in Japan.) The term ``as follows'' here indicates the contents from Part I to Ultraviolet-visible Reference SpectraintheSupplementIItotheJapanesePharmacopoeiaFourteenthEdition(pp.1669 – 1866).
Transcript
The Ministry of Health, Labour andWelfare Ministerial Notiˆcation No. 461
In accordance with the provisions of Article 41, Paragraph 1 of the PharmaceuticalAŠairs Law (Law No. 145, 1960), we hereby revise a part of the Japanese Phar-macopoeia (Ministerial Notiˆcation No. 111, 2001) as follows, and the revisedJapanese Pharmacopoeia shall come into eŠect on January 1, 2005, [including dele-tion from O‹cial Monographs fro Part II in The Japanese Pharmacopoeia, Four-teenth Edition of the articles of Absorbent Cotton, Puriˆed Absorbent Cotton,Sterile Absorbent Cotton, Sterile Puriˆed Absorbent Cotton and Absorbent Gauzeand Sterile Absorbent Gauze (hereinafter referred to as ``sanitary materials'')]. Provi-so: With respect to the drugs which are included in the Japanese Pharmacopoeia(hereinafter referred to as ``the old Japanese Pharmacopoeia'') [limited to thoseincluded in the Japanese Pharmacopoeia whose standards are changed with thisnotiˆcation published (hereinafter referred to as ``the new Japanese Phar-macopoeia'')] and those which are approved as of January 1, 2005 pursuant to theprovisions of Article 14, Paragraph 1 of this Law (including cases where it shall applymutatis mutandis under Article 23 of this Law; the same hereinafter) [including thedrugs designated as those exempted from approval (hereinafter referred to as ``thedrugs exempted from approval'') among the drugs etc. designated by the Minister ofHealth, Labour and Welfare as those exempted from manufacturing or importapproval pursuant to the provisions of Article 14, Paragraph 1 of the PharmaceuticalAŠairs Law (Ministerial Notiˆcation No. 104, 1994), the standards established in theold Japanese Pharmacopoeia (limited to the standards for the relevant drugs) shall berecognized, up to June 30, 2006, as the standards established in the new JapanesePharmacopoeia. With respect the drugs which are included in the new JapanesePharmacopoeia (excluding those which are included in the old Japanese Phar-macopoeia) and those which are approved as of January 1, 2005 pursuant to theprovisions of Article 14, Paragraph 1 of this Law (including the drugs exempted fromapproval), the drugs may be treated, up to June 30, 2006, as those which are notincluded in the new Japanese Pharmacopoeia. Further, sanitary materials may betreated, up to September 30, 2006, under the previous regulation.
Hidehisa OtsujiThe Minister of Health, Labour and Welfare
December 28, 2004
(The texts referred to by the term ``as follows'' are omitted here. All of them aremade available for public exhibition at the Evaluation and Licensing Division, Phar-maceutical and Medical Safety Bureau, Ministry of Health, Labour and Welfare, ateach Regional Bureau of Health and Welfare and at each Prefectural O‹ce in Japan.)
The term ``as follows'' here indicates the contents from Part I to Ultraviolet-visible ReferenceSpectraintheSupplementIItotheJapanesePharmacopoeiaFourteenthEdition(pp.1669 – 1866).
CONTENTS
Preface ...................................................... iSupplement II to The Japanese Pharmacopoeia,Fourteenth Edition, Part I................. 1669–1754
General Tests, Processes and Apparatus ... 16696. Bacterial Endotoxins Test................. 1669
39. Nuclear Magnetic ResonanceSpectroscopy ................................. 1669
47. Pyrogen Test ................................. 167252. Residue on Ignition Test .................. 167253. Speciˆc Surface Area Determination... 167354. Sterility Test .................................. 167370. Reference Standards; Reagents, Test Solu-
tions; Standard Solutions for VolumetricAnalysis; Standard Solutions; MatchingFluids for Color; Optical Filters forWavelength and Transmission RateCalibration; and Measuring Instruments,Appliances .................................... 1677(1) Reference Standards ................. 1677(2) Reagents, Test Solutions............ 1677(3) Standard Solutions for Volumetric
Analysis ................................. 168574. Powder Particle Density
Based on Molecular BiologicalMethod ........................................ 1833
25. Solid and Particle Densities .............. 183526. Total Protein Assay ........................ 1836
Index.................................................... 1841Index in Japanese.................................... 1857
ii
Preface
The Fourteenth Edition of the Japanese Phar-macopoeia was promulgated on March 30, 2001 byMinisterial Notiˆcation No. 111 of the Ministry ofHealth, Labour and Welfare. To keep pace withprogress in medical and pharmaceutical sciences, inNovember 2001, the Council, at a meeting of theCommittee on Japanese Pharmacopoeia (JP), estab-lished the basic principles for the preparation of theJP Fifteenth Edition, setting out the characteristicsand roles of the JP, the deˆnite measures for therevision, the date of the revision, and the organizationof the Subcommittee on JP.
At the above meeting, the following ``ˆve pillars''were established as the basic principles of the JP:Making it more substantial by including all drugswhich are important from the viewpoint of health careand medical treatment; Making prompt partialrevision as necessary and facilitating smooth adminis-trative operation; Promoting international harmoni-zation; Ensuring transparency regarding the revisionand dissemination to the public of the JP; andPositively introducing contemporary analytical testsand developing reference standards. It was decided atthe meeting that each panel set up under the Subcom-mittee on JP should make eŠorts, on the basis of theseprinciples, to ensure that the JP is used more eŠective-ly in the ˆelds of health care and medical treatment bytaking appropriate measures, including getting theunderstanding and cooperation of other parties con-cerned.
The JP should comprise an o‹cial standard beingrequired to assure the quality of drugs in this countryin response to the progress in science and technologyand clinical demands at the time, it should deˆne thestandards for speciˆcations as well as the methods oftests to assure the overall quality of all drugs inprinciple, and it should have a role in clarifying thecriteria for quality of drugs which are recognized to beimportant from the viewpoint of medical treatment.
At the same time, it was agreed that the JP shouldbe prepared with the aid of the knowledge andexperience of many persons involved in the phar-maceuticals, that it should have the characteristics ofan o‹cial standard, which might be widely used by allparties concerned, that it should provide informationand understanding about the quality of drugs to thepublic, and that it should be conducive to smooth andeŠective government control of the quality of drugs,
and to securing and maintaining international con-sistency.
It was also agreed that JP articles should coverdrugs which are important from the viewpoint ofhealth care and medical treatment, clinical results andfrequency of use, as soon as possible after they reachthe market.
It was also decided to make a deˆnite rule for selec-tion of articles by clarifying the standards for selec-tion. The rule was shown in the verdict ``What thefuture Japanese Pharmacopoeia should be'' by thePharmaceutical AŠairs and Food Sanitation Council(PAFSC) on December 2002. The JP FifteenthEdition was decided to be slated for completion inApril 2006.
The panels on JP was reorganized into the followingeleven panels in accordance with the recommendationof the PAFSC: Panel on Planning and Revisions;Panel on Nomenclature for Pharmaceuticals; Panelon Excipients; Panel on Physico-chemical Tests; Panelon Medicinal Chemicals; Panel on BiologicallyDerived Drugs; Panel on Biological Tests; Panel onAntibiotics; Panel on Crude Drugs; Provisional Panelon Planning and Revisions and Panel on Phar-macopoeial Harmonization (PDG), followed by theestablishment of new panels: Panel on Water forPharmaceutical Preparations, Panel on JP ReferenceStandards, and three working groups under Panel onMedicinal Chemicals to expedite discussion of revisiondrafts of drug monographs.
In the Committee on Japanese Pharmacopoeia,Mitsuru Uchiyama took the role of chairman fromJanuary 2001 to December 2002, Tadao Terao fromJanuary to June 2003, and Takao Hayakawa fromJuly 2003 to December 2004.
It was decided that the JP will be revised not onlyevery ˆve years, in line with the basic principles for thepreparation of the JP Fifteenth Edition, but also asnecessary to take account of recent progress of scienceand in the interests of international harmonization.
In accordance with the revision principles, thepanels continued discussions on selection of articles,and revisions for General Notices, General Rules forPreparations, General Tests, and monographs ondrugs, and the supplement I to JP 14 was promulgatedon December 2002. After the promulgation of thesupplement I, the panels continued discussions to takeaccount of the progress of science and international
iiii Supplement II, JPXIVPreface
harmonization.Draft revisions covering subjects in General Rules
for Crude Drugs, General Tests, and monographs ondrugs, for which discussions were ˆnished betweenMarch 2002 and December 2003, were prepared for asupplement to the book. They were examined by theCommittee on Japanese Pharmacopoeia in September2004, followed by the PAFSC in December 2004, andthen submitted to the Minister of Health, Labour andWelfare.
Numbers of discussions in the panels to preparesupplement drafts were as follows: Panel on Planningand Revisions, 9 times; Panel on Nomenclature forPharmaceuticals, 10 times; Panel on Excipients, 11times; Panel on Physico-chemical Tests, 30 times;Panel on Medicinal Chemicals (including the workinggroups), 24 times; Panel on Biologically DerivedDrugs, 11 times; Panel on Biological Tests, 10 times;Panel on Antibiotics, 19 times; Panel on Crude Drugs,19 times; Provisional Panel on Planning and Revi-sions, 12 times; Panel on Pharmacopoeial Harmoniza-tion (PDG), 9 times; Panel on Water for Pharmaceuti-cal Preparations, 2 times; Panel on JP ReferenceStandards, 3 times.
It should be noted that in the preparation of thedrafts for the Supplement I, generous cooperation wasgiven by the Technical Committee of the Pharmaceuti-cal Manufacturer's Association of Tokyo and ofOsaka, the Crude Drugs Association of Tokyo, theJapan Pharmaceutical Excipients Council, theFederation of Crude Drugs Associations of Japan, theJapan Antibiotics Research Association, the JapanFlavor and Fragrance Manufacturer's Association,the Japan Medical Plants Federation, the JapanPharmaceutical Manufacturer's Association, theJapanese Society of Hospital Pharmacists, the JapanPharmaceutical Association, and the Japan OilseedProcessors Association.
In consequence of this revision, the JP FourteenthEdition carries 907 articles in Part I owing to theaddition of 27 articles and the deletion of 1 article; and484 articles in Part II owing to the addition of 12articles and the deletion of 9 articles.
The principles of description and the salient pointsof the revision in this volume are as follows:
1. The Supplement II to JP Fourteenth Editioncomprises the following items, in order: Notiˆcationof the Ministry of Health, Labour and Welfare;Contents; Preface; followed by General Tests, Proc-esses and Apparatus; monographs on drugs in Part I,and General Rules for Crude Drugs; monographs ondrugs in Part II, then followed by Ultraviolet-visual
Reference Spectra in Part I; Infrared ReferenceSpectra and Ultraviolet-visible Reference Spectra inPart II; General Information, and as an appendix, aCumulative Index containing references to the mainvolume, the supplement I and the supplement II.
2. The articles in General Tests, Processes andApparatus, Monographs on Drugs, Infrared Refer-ence Spectra and Ultraviolet-visible Reference Spectraare respectively placed in alphabetical order.
3. The following items in each monograph are putin the order shown below, except that unnecessaryitems are omitted depending on the nature of the drug:(1) English title(2) Commonly used name(s)(3) Latin title (only for Crude Drugs)(4) Title in Japanese(5) Structural formula or empirical formula(6) Molecular formula and molecular mass(7) Chemical name(8) Origin(9) Limits of the content of the ingredient(s)
and/or the unit of potency(10) Labeling requirements(11) Method of preparation(12) Description(13) Identiˆcation tests(14) Speciˆc physical and/or chemical values(15) Purity tests(16) Loss on drying, loss on ignition, and/or water(17) Residue on ignition, total ash, and/or acid-in-
soluble ash(18) Tests being required for pharmaceutical prepa-
rations and other special tests(19) Isomer ratio(20) Assay or the content of the ingredient(s)(21) Containers and storage(22) Expiration date(23) Others
4. In each monograph on a drug, the followingphysical and chemical values representing the proper-ties and quality of the drug are given in the order indi-cated below, except that unnecessary items are omitteddepending on the nature of the drug:(1) Alcohol number(2) Absorbance(3) Congealing point(4) Refractive index(5) Osmolarity(6) Optical rotation(7) Viscosity(8) pH(9) Speciˆc gravity
5. Identiˆcation tests comprise the followingitems, which are generally put in the order given be-low:(1) Coloration reactions(2) Precipitation reactions(3) Decomposition reactions(4) Derivatives(5) Visible, ultraviolet or infrared spectra(6) Special reactions(7) Cations(8) Anions
6. Purity tests comprise the following items, whichare generally put in the order given below, except thatunnecessary items are omitted depending on thenature of the drug:(1) Color(2) Odor(3) Clarity and/or color of solution(4) Acidity or alkalinity(5) Acid(6) Alkali(7) Chloride(8) Sulfate(9) Sulˆte
(32) Silver(33) Alkaline earth metals(34) Arsenic(35) Foreign matter(36) Related substances(37) Residual solvent(38) Other mixtures(39) Readily carbonizable substances
7. The following items of the General Tests, Proc-esses and Apparatus were partially revised:
Bacterial Endotoxins TestNuclear Magnetic Resonance Spectroscopy (1H)Pyrogen TestResidue on Ignition TestSpeciˆc Surface Area DeterminationSterility Test
8. The following test was added to the GeneralTests, Processes and Apparatus:
Powder Particle Density Determination
9. The following Reference Standard was deleted:Digitalis
11. English and Latin titles of drugs are derived,in principle, from International NonproprietaryNames (INN) for Pharmaceutical Substances recom-mended by the World Health Organization. Japanesetitles are derived from the Japanese version of thisbook. The chemical names are based on the rules ofthe International Union of Pure and Applied Chemis-try (IUPAC).
12. Molecular formulas of organic compoundsbegin with C and then H, followed by other involvedelements in the alphabetical order of the symbols ofthe elements.
13. Structural formulas of drugs represent, as faras possible, steric conˆgurations.
14. Test procedures in monographs in Part I are,in principle, written in full even in correspondingmonographs in Part II, and vice versa. The test proce-
iviv Supplement II, JPXIVPreface
dures in monographs for preparations are also writtenin full even within the same part, except in the mono-graphs for preparations having a correspondingmonograph of their principal material substances.
15. The following articles were deleted fromO‹cial MonographPart I
*Chairman, Committee on JP**Acting Chairman, Committee on JP
16691669
General Tests, Processesand Apparatus
Change the introduction to read:
General Tests, Processes and Apparatus includes commonmethods for tests and other articles related to them. Unlessotherwise speciˆed, the procedures for absorbance determi-nation, absorbance ratio determination, acid-neutralizingcapacity determination of gastrointestinal medicines,alcohol number determination, ammonium determination,arsenic determination, atomic absorption spectrophotomet-ry, test for bacterial endotoxins, boiling point determina-tion, distilling range determination, chloride determination,conductivity measurement, congealing point determination,test for content uniformity, determination of bulk andtapped densities, digestion test, disintegration test, dissolu-tion test, endpoint detection in titrimetry, ‰ame coloration,‰uorometry, foreign insoluble matter test for injections, gaschromatography, heavy metals determination, test for glasscontainers for injections, infrared spectrophotometry,insoluble particulate matter test for injections, insolubleparticulate matter test for ophthalmic solutions, iron deter-mination, liquid chromatography, loss on drying determina-tion, loss on ignition determination, mass variation test,melting point determination, test for metal particles inophthalmic ointments, methanol determination, methoxylassay, microbial assay for antibiotics, test for microbiallimit, test for microbial limit for crude drugs, mineral oildetermination, nitrogen determination, nuclear magneticresonance spectroscopy, optical rotation determination,osmolarity determination, oxygen ‰ask combustion method,paper chromatography, particle size distribution test forpreparations, pH determination, test for plastic containers,powder particle density determination, powder particle sizedetermination, test for pyrogen, qualitative test, test forreadily carbonizable substances, refractive index determina-tion, residual solvents test, residue on ignition determina-tion, test for rubber closure for aqueous infusions, speciˆcgravity and density determination, speciˆc surface areadetermination, test for sterility, sulfate determination,thermal analysis, thin-layer chromatography, test for totalorganic carbon, viscosity determination, vitamin A assay,test for volatile contaminants in ethanol, water determina-tion, and X-ray powder diŠraction are performed as directedin the corresponding articles under the General Tests, Proc-esses and Apparatus. The tests for melting point of fats,congealing point of fatty acids, speciˆc gravity, acid value,saponiˆcation value, ester value, hydroxyl value, unsaponiˆ-able matter and iodine value of fats and fatty oils areperformed as directed in the corresponding items under theFats and Fatty oils Test, and the tests for foreign matter and
loss on drying, total ash, acid-insoluble ash, extract content,essential oil content of crude drugs are performed as directedin the corresponding items under the Crude Drugs Test.
6. Bacterial Endotoxins TestChange the (ii) Interpretation of (2) Limit testunder Gel-clot techniques to read:
Gel-clot techniques(2) Limit test(ii) InterpretationThe test is valid when both replicates of solutions B and C
are positive and those of solution D are negative.The sample meets the endotoxin limit requirement of the
test when a negative result is found for both replicates ofsolution A.
Repeat the test in duplicate when the test results are posi-tive for one test but negative for the other one. The samplemeets the endotoxin limit requirement of the test when anegative result is found for both replicates of solution A inthe repeat test.
The sample does not meet the endotoxin limit requirementof the test when a positive result is found for both replicatesof the solution A at a dilution equal to the MVD. If the testis positive for the sample at a dilution less than the MVD,the test may be performed at a dilution not greater than theMVD.
39. Nuclear Magnetic ResonanceSpectroscopy (1H)
Change to read:
39. Nuclear Magnetic ResonanceSpectroscopy
Nuclear magnetic resonance (NMR) spectroscopy is basedon the phenomenon that speciˆc radio frequency radiation isabsorbed by magnetic nuclei in a sample placed in a magnet-ic ˆeld; target nuclei are 1H, 13C, 15N, 19F, 31P, etc. Thesenuclei have intrinsic spin angular momentum, of which themagnitude is given by I (I＋1)/h/2p, where I is the spinquantum number and is integral or half-integral (I=1/2 for1H and 13C). When the magnetic nuclei are placed in a mag-netic ˆeld, they are oriented in 2I＋1 possible orientations
16701670 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
corresponding to 2I＋1 equally spaced energy levels (twoenergy levels for 1H and 13C). The transition between twosuccessive quantized energy levels corresponding to adjacentorientations can be induced by electromagnetic radiationwith a suitable frequency. The precise relation between theˆeld strength and the resonant frequency n is given by
n＝g･H0
2pwhere H0 is the strength of the applied external magneticˆeld and g is the gyromagnetic ratio, a constant characteriz-ing a particular isotope. The absorption of radiation (NMRsignal) can occur only when the irradiating radio frequencysatisˆes the resonance condition. Since the absorptioncoe‹cient (the transition probability) does not depend onthe environment in which the nuclei are located, the intensityis basically proportional to the number of nuclei. The excessspins shifted to the higher energy levels by the transitionprocess return to the thermal equilibrium state at variousrates determined by a characteristic time constant (known asthe relaxation time).
A nucleus is shielded from the applied magnetic ˆeld bythe electrons belonging to its own atom and to the molecule.Therefore nuclei in diŠerent environments are shielded todiŠerent extents and resonate at diŠerent frequencies. ThediŠerence in resonance frequencies is deˆned as chemicalshift (d), which is independent of the strength of the magnet-ic ˆeld, and is given by
d＝nS－nR
nR＋dR
where,nS: The resonance frequency of the observed signal,nR: The resonance frequency of the reference signal,nR: The chemical shift of the reference signal (in the case
of the value not being 0).
The chemical shifts are normally expressed in ppm, adimensionless unit, by assuming the chemical shift of thereference compound as 0 ppm. When the chemical shift ofthe reference compound is not assumed to be 0 ppm, chemi-cal shifts of samples are corrected accordingly.
In addition to the shielding due to electrons, the nucleus issubjected to eŠects due to the spin orientations of othermagnetic nuclei through chemical bonds, resulting in anadditional splitting of the signal. The spacing between twoadjacent components of the signal is known as the spin-spincoupling constant (J). Coupling constants are measured inhertz and are independent of the strength of the externalmagnetic ˆeld. The increased number of interacting nucleiwill make the multiplet pattern more complex.
From the NMR spectrum the following four parameterscan be obtained: chemical shift, spin-spin coupling constant,resonance intensity (intensities of 1H are proportional to thenumber of nuclei and those of 13C and others are susceptibleto the nuclear Overhauser eŠect (NOE) and relaxation) andrelaxation time. These parameters are useful for structuraldetermination, identiˆcation and quantitative analysis ofmolecules. Spin decoupling, NOE, and two-dimensionalNMR techniques are also available for structural analysis.
SpectrometerThere are two types of spectrometers.(1) Fourier transform NMR (FT-NMR) spectrometers
(Figure 1)Target nuclei are simultaneously excited in all frequency
range of the nuclei by means of an intense radio frequencypulse. The FID (free induction decay) after the pulse isdetected, which is a time domain signal, is converted to afrequency domain spectrum by Fourier transformation.Number of data points suitable for the spectral range, ‰ipangle, acquisition time, delay time and number of scansshould be set appropriately.
Recently FT-NMR is commonly used because of its highsensitivity and various advanced applications.
In the case of the CW method, a spectrum is obtained bysweeping the radio frequency or magnetic ˆeld continuouslyover the frequency range of the nuclei being observed.
Fig. 2 CW-NMR spectrometer
MeasurementPrior to measurements, the sensitivity and resolution of
the instrument must be adjusted to the optimum levels usinga standard sample (ethylbenzene, 1,2-dichlorobenzene oracetaldehyde) dissolved in an appropriate NMR solvent.
(1) The sample dissolved in a suitable solvent is trans-
16711671Supplement II, JPXIV General Tests, Processes and Apparatus
ferred into an NMR tube. The reference compound can beadded directly to the sample solution (internal reference), ora sealed capillary tube containing the reference compoundcan be inserted into the NMR tube (external reference). Thesample solutions should be completely homogeneous. Inparticular, solid contaminants should be removed in order toobtain good spectra. Various deuterated NMR solvents arecommonly used for NMR measurement and the followingpoints should be considered in selecting an appropriatesolvent: (i) The solvent signals do not overlap with thesample signals. (ii) The sample must be soluble in the solventselected. (iii) The solvent does not react with the sample.Furthermore, it should be noted that chemical shifts candepend upon the solvent employed, sample concentrationand deuterium ion concentration, and that viscous solutionsusually give rather broad, poorly resolved spectra.
(2) For the reference standards use the reagents fornuclear magnetic resonance spectroscopy. For 1H and 13Cspectra, tetramethylsilane (TMS) is usually used as thereference compound for samples dissolved in organicsolvents. For samples dissolved in deuterium oxide, sodium2,2-dimethyl-2-silapentane-5-sulfonate (DSS) or sodium 3-(trimethylsilyl)propionate (TSP) is used. For other nuclei,nitromethane, trichloro‰uoromethane and phosphoric acidare used as reference compounds for 15N, 19F and 31P,respectively. Furthermore, chemical shifts of residual pro-tons in deuterated solvents and 13C in the solvent instead of areference compound can be used for 1H and 13C NMR.
Record of apparatus and measurement conditionsType of instrument, frequency, solvent, temperature,
sample concentration, reference compound, experimentaltechnique, etc. should be recorded to allow appropriatecomparison of spectra, because NMR spectra depend on themeasurement conditions.
IdentiˆcationThe sample solution is prepared and tested by the method
directed in each monograph. Usually in the case of 1H NMR,the sample is identiˆed by the following method.
(1) Identiˆcation by the use of chemical shift, multiplic-ity and relative intensity
When chemical shifts, multiplicities and relative intensi-ties of signals are deˆned, the sample can be identiˆed asbeing the same substance when all chemical shifts, multiplic-ities and relative intensities are the same as those prescribed.
(2) Identiˆcation by the use of a Reference StandardMeasurement conditions should be the same as those used
in the case of the Reference Standard. When the spectra of asample and the Reference Standard exhibit the samemultiplicities and relative intensities of signal at the samechemical shifts, the sample can be identiˆed as being thesame substance as the Reference Standard.
Experimental techniques of 1H and 13C NMR spectroscopyNMR spectroscopy includes one-, two- and multi-dimen-
sional techniques, which are used for various purposes.Spin decoupling, and NOE are available in one-dimen-
sional 1H spectroscopy. Spin decoupling can assign coupling
correlations. As NOE can observe correlations amongspatially proximate protons, the conˆguration and theconformation can be analyzed.
Broadband decoupling, INEPT and DEPT are usuallyapplied in one-dimensional 13C spectroscopy. The broad-band decoupling technique simpliˆes a spectrum andachieves enhancement of sensitivity. INEPT (insensitivenuclei enhanced by polarization transfer) and DEPT (distor-tionless enhancement of polarization transfer) enhance thesensitivity of 13C by means of polarization transfer fromdirectly bonded 1H with a large magnetic moment. They canbe applied to identify primary, secondary, tertiary orquarternary carbon.
Two-dimensional spectroscopy can observe all correlationpeaks between nuclei through spin-spin coupling or NOE ina single experiment, and there are many techniques forhomonuclear and heteronuclear measurements. Representa-tive techniques are described below.
COSY (2D correlation spectroscopy), HOHAHA(homonuclear Hartmann-Hahn spectroscopy) or TOCSY(total correlation spectroscopy): Correlation between pro-tons through scalar spin-spin coupling is obtained andintramolecular connectivities of hydrogen atoms arerevealed.
NOESY (2D nuclear Overhauser enhancement andexchange spectroscopy): NOE is measured by a two-dimen-sional technique. Approximate distances between spatiallyproximate hydrogen atoms are obtained to analyze the three-dimensional structure.
INADEQUATE (incredible natural abundance doublequantum transfer experiment): Although this technique isinsensitive because it involves double quantum transfer by13C-13C scalar coupling in a sample with natural isotopicabundance, the connectivities of all neighboring 13C nucleican be obtained to analyze the carbon skeleton.
HMQC (heteronuclear multiple quantum coherence): Thistechnique observes correlations between 1H and 13C withdirect spin-spin coupling using 1H detection and revealsintramolecular chemical bonds between hydrogen andcarbon atoms.
HMBC (heteronuclear multiple bond connectivity): Thistechnique observes correlations between 1H and 13C withlong range spin-spin coupling using 1H detection and revealsintramolecular connectivities of hydrogen and carbonatoms.
There are many other techniques such as DQF-COSY(double quantum ˆltered COSY) and HSQC (heteronuclearsingle quantum coherence). Furthermore, multidimensionalNMR techniques are used to analyze macromolecules.
16721672 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
47. Pyrogen TestChange to read:
The Pyrogen Test is a method to test the existence ofpyrogens by using rabbits.
Test animalsUse healthy mature rabbits, each weighing not less than
1.5 kg, which have not lost body mass when kept on aconstant diet for not less than one week. House the rabbitsindividually in an area free from disturbances likely to excitethem. Keep the temperature of the area constant between209C and 279C for at least 48 hours before and throughoutthe test. Before using a rabbit that has not previously beenused for a pyrogen test, condition it 1 to 3 days prior to thetest by conducting a sham test omitting the injection. Do notuse a rabbit for pyrogen tests more frequently than onceevery 48 hours, or after it has been given a test sample thatwas adjudged pyrogen-positive or that contained an antigenpresent commonly in the test sample to be examined.
Apparatus, instruments(1) Thermometer—Use a rectal thermometer or temper-
ature-measuring apparatus with an accuracy of ±0.19C orless.
(2) Syringe and injection needle—Depyrogenate thesyringes and needles in a hot-air oven using a validated proc-ess, usually by heating at 2509C for not less than 30 minutes.Sterilized syringes with needles are also available providedthat they have been validated to assure that they are free ofdetectable pyrogens and do not interfere with the test.
Test procedures(1) Quantity of injection—Unless otherwise speciˆed,
inject 10 mL of the sample per kg of body mass of eachrabbit.
(2) Procedure—Perform the test in a separate area at anenvironmental temperature similar to that of the roomwherein the animals were housed and free from disturbanceslikely to excite them. Withhold food from the rabbits forseveral hours before the ˆrst record of the temperature andthroughout the testing period. The test animals are usuallyrestrained with loosely ˆtting neck stocks that allow therabbits to assume a natural resting posture. Determine thetemperature of each rabbit by inserting the thermometer ortemperature-measuring probe into the rectum of the testanimal to a constant depth within the range of 60 mm to90 mm. The ``control temperature'' of each rabbit is themean of two temperature readings recorded for that rabbitat an interval of 30 min in the 40 min immediately precedingthe injection of the sample to be examined. Rabbits showinga temperature variation greater than 0.29C between the twosuccessive temperature readings or rabbits having an initialtemperature higher than 39.89C are withdrawn from thetest.
Warm the test solution to a temperature of 37±29Cbefore injection, and inject the solution slowly into themarginal vein of the ear of each rabbit over a period not
exceeding 10 min. Hypotonic test sample may be madeisotonic by the addition of pyrogen-free sodium chloride.Record the temperature of each rabbit during a period of3 hours after the injection, taking the measurements atintervals of not more than 30 min. The diŠerence betweenthe control temperature and the maximum temperature ofeach rabbit is taken to be the rise in body temperature.Consider any temperature decreases as zero rise.
Interpretation of resultsThe test is carried out on a group of three rabbits and the
result is judged on the basis of the sum of the three tempera-ture rises. Repeat if necessary on further groups of three rab-bits to a total of three groups, depending on the resultsobtained. If the summed response of the ˆrst group does notexceed 1.39C, the sample is judged to be pyrogen-negative.If the summed response exceeds 2.59C, the sample is judgedto be pyrogen-positive. If the summed response exceed1.39C but does not exceed 2.59C, repeat the test on anothergroup of three rabbits. If the summed response of the ˆrstand second group does not exceed 3.09C, the sample isjudged to be pyrogen-negative. If the summed response ofthe 6 rabbits exceeds 4.29C, the sample is judged to bepyrogen-positive. If the summed response exceeds 3.09C butdoes not exceed 4.29C, repeat the test on one more group ofthree rabbits. If the summed response of the 9 rabbits doesnot exceed 5.09C, the sample is judged to be pyrogen-negative. If the summed response exceeds 5.09C, the sampleis judged to be pyrogen-positive.
When the test sample is judged to be pyrogen-negative,the sample passes the pyrogen test.
52. Residue on Ignition TestChange the Procedure to read:
ProcedurePreviously ignite a suitable crucible (silica, platinum,
quartz or porcelain) at 600±509C for 30 minutes, and weighaccurately after cooling in a desiccator (silica gel or othersuitable dessicant).
Take the sample of the amount directed in the mono-graph, transfer into the ignited crucible, and weigh accurate-ly. When the quantity of the sample to be taken is indicatedin a volume, pipet exactly the amount directed in themonograph and transfer into the above crucible. Whendirected as ``after evaporating,'' heat properly to evaporatethe solution.
Moisten the sample with a small amount of sulfuric acid,usually 1 mL, then heat slowly at a temperature as low aspracticable until the sample is completely carbonized, andcool. Moisten again with a small amount (usually 1 mL) ofsulfuric acid, heat gently until white fumes are no longerevolved, and ignite at 600±509C until the residue is com-pletely incinerated. Ensure that ‰ames are not produced atany time during the procedure. Cool the crucible in a desic-cator (silica gel or other suitable dessicant), and reweighaccurately to calculate the percentage of residue.
16731673Supplement II, JPXIV General Tests, Processes and Apparatus
Unless otherwise speciˆed, if the amount of the residue soobtained exceeds the limit speciˆed in the individual mono-graph, repeat the moistening with sulfuric acid, heating andignition as before until constant mass is attained or until thepercentage of residue complies with the limit in the individ-ual monograph.
53. Speciˆc Surface AreaDetermination
Change to read the text up to the paragraphheaded ``Method 1 Dynamic Flow Method'' asfollows:
Speciˆc Surface Area Determination is a method to deter-mine the speciˆc surface area (the total surface area ofpowder per an unit mass) of a powdered pharmaceuticalpreparation by using the gas adsorption method. The gasadsorption method is a method for measuring the amount ofgas adsorbed on the surface of a powder sample as a func-tion of the pressure of the adsorbate gas, and the measure-ments are usually performed at the boiling point of liquidnitrogen (－1969C).
When the gas is physically adsorbed by the powdersample, the following relationship holds when P/P0 is in therange of 0.05 to 0.30 for pressure P of the adsorbate gas inequilibrium for the volume of gas adsorbed, Va.
1
VaØ P0
P－1»
＝(C－1)VmC
×PP0＋
1VmC
P: Partial vapor pressure of adsorbate gas in equilibrium(kPa)
P0: Saturated vapor pressure of the adsorbate gas at－1969C (kPa)
Va: Volume of gas adsorbed at equilibrium (mL)Vm: Volume of gas adsorbed in a monolayer (mL)C: Dimensionless constant relating to the enthalpy of
adsorption and condensation of the adsorbate gasThe speciˆc surface area, S, is determined from Vm, the
volume of gas adsorbed in a monolayer on the sample.
S＝Vm×N×am×22400
S: Speciˆc surface area (m2/g)N: Avogadro constant 6.022×1023/mola: EŠective cross-sectional area of one adsorbate molecule
(m2)N20.162×10－18
Kr: 0.195×10－18
m: Mass of the test powder (g)Speciˆc surface area is generally expressed in unit of m2/g.Either of the methods described below can be used to
measure the gas adsorption.
54. Sterility TestChange to read:
Test for sterility is the method to establish the presence orabsence of viable microorganisms (bacteria and fungi) usingthe deˆned culturing method. Unless otherwise speciˆed, thetest is carried out by I. Membrane ˆltration method or II.Direct inoculation method. Water, reagents, test solutions,equipment, materials and all other requisites for the testshould be pre-sterilized. The test for sterility is carried outunder aseptic conditions. In order to achieve such condi-tions, the test environment has to be adapted to the way inwhich the sterility test is performed. The precautions takento avoid contamination are such that they do not aŠect anymicroorganisms which are to be revealed in the test. Theworking conditions in which the tests are performed aremonitored regularly by appropriate sampling of the workingarea and by carrying out appropriate controls.
Media and rinsing ‰uidsFluid thioglycolate medium, soybean-casein digest
medium are used, unless otherwise speciˆed. When it isdi‹cult to use ‰uid thioglycolate medium due to turbidity orviscosity of samples, alternative thioglycolate medium canbe used, provided it is heated on a water bath just prior touse and incubated under anaerobic conditions. Otherproducts of suitable quality yielding similar formulationsmay be used according to the indications on the label.
(1) Fluid thioglycolate mediumL-Cystine 0.5 gAgar 0.75 gSodium chloride 2.5 gGlucose, monohydrate/anhydrate 5.5/5.0 gYeast extract (water-soluble) 5.0 gPancreatic digest of casein 15.0 gSodium thioglycolate or 0.5 g
Thioglycolic acid 0.3 mLResazurin sodium solution (1 in 1000),
freshly prepared 1.0 mLWater 1000 mL
(pH after sterilization 7.1±0.2)Mix the L-cystine, agar, sodium chloride, glucose, water-
soluble yeast extract and pancreatic digest of casein with thewater, and heat until solution is eŠected. Dissolve thesodium thioglycolate or thioglycolic acid in the solution and,if necessary, add sodium hydroxide TS so that, after sterili-zation, the solution will have a pH of 7.1±0.2. If ˆltration isnecessary, heat the solution again without boiling and ˆlterwhile hot through moistened ˆlter paper. Add the resazurinsodium solution, mix and place the medium in suitablevessels which provide a ration of surface to depth of mediumsuch that not more than the upper half of the medium hasundergone a color change indicative of oxygen uptake at theend of the incubation period. Sterilize using a validatedprocess. Store the medium at a temperature between 2 –259C. If more than the upper one-third of the medium hasacquired a pink color, the medium may be restored once by
16741674 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
heating the containers in a water-bath or in free-‰owingsteam until the pink color disappears and cooling quickly,taking care to prevent the introduction of non-sterile air intothe container.
(2) Alternative thioglycolate mediumL-Cystine 0.5 gSodium chloride 2.5 gGlucose, monohydrate/anhydrate 5.5/5.0 gYeast extract (water-soluble) 5.0 gPancreatic digest of casein 15.0 gSodium thioglycolate or 0.5 g
Thioglycolic acid 0.3 mLWater 1000 mL
(pH after sterilization 7.1±0.2)The methods for preparation follow those of ‰uid
thioglycolate medium.(3) Soybean-casein digest medium
(pH after sterilization 7.3±0.2)Mix all the ingredients and heat until solution is eŠected.
If necessary, add sodium hydroxide TS so that, after sterili-zation, the solution will have a pH of 7.3±0.2. Filter, ifnecessary, to clarify, distribute into suitable vessels andsterilize using a validated process. Store at a temperaturebetween 2 – 259C in a sterile container.
(4) Rinsing ‰uidsMeat or casein peptone 1.0 gWater 1000 mL
(pH after sterilization 7.1±0.2)Dissolve animal tissue or casein peptone in water and
adjust the pH of the solution so that, after sterilization, itwill show 7.1±0.2. Filter, if necessary, to clarify, distributeinto suitable vessels and sterilize using a validated process.Store at a temperature between 2 – 259C in a sterilecontainer.
To rinsing ‰uid to be used for antibiotics or pharmaceuti-cal products containing an antimicrobial agent, a suitableneutralizer or inactive agent at concentration shown to beappropriate in the validation of the test can be added. Torinsing ‰uid to be used for oils, oily solutions, ointments orcreams, suitable emulsifying agent at a concentration shownto be appropriate in the validation of the test, for examplepolysorbate 80 at a concentration of 10 g/L can be added.
Suitability of mediaThe media used comply with the following tests, carried
out before or in parallel with the test on the product to beexamined.
(1) Sterility of mediaConˆrm the sterility of each sterilized batch of medium by
incubating a portion of the media at the speciˆed incubationtemperature for 14 days. No growth of microorganismsoccurs.
(2) Growth promotion testTest each batch of ready-prepared medium and each batch
(lot)of medium prepared either from dehydrated medium orfrom ingredients1. Inoculate a small number (not more than100 CFU) of microorganism listed in Table 1 or other strainsconsidered to be equivalent to these strains in containers ofeach medium. Each of the test organisms should showclearly visible growth in all inoculated media within 3 daysfor bacteria and within 5 days for fungi.
Table 1. Microorganisms for growth promotion test andthe validation test
Seed lot culture maintenance techniques (seed-lot systems)are used so that the viable microorganisms used for inocula-tion are not more than ˆve passages removed from theoriginal master seed-lot.
EŠective period of mediaIf prepared media are stored in unsealed containers, they
can be used for one month, provided that they are tested forgrowth promotion within two weeks of the time of use andthat color indicator requirements are met. If stored in tightcontainers, the media can be used for one year, providedthat they are tested for growth promotion within 3 monthsof the time of use and that the color indicator requirementsare met.
Validation testThe validation may be performed simultaneously with the
Test for sterility of the product to be examined in thefollowing cases.a) When the test for sterility has to be carried out on a new
product.
16751675Supplement II, JPXIV General Tests, Processes and Apparatus
b) Whenever there is a change in the experimental condi-tions of the test.
Carry out the test as described below under Test for sterili-ty of the product to be examined using exactly the samemethods except for the following modiˆcations.Membrane ˆltration After transferring the content of thecontainer or containers to be tested to the membrane add aninoculum of a small number of viable micro-organisms (notmore than 100 CFU) to the ˆnal portion of sterile diluentused to rinse the ˆlter.Direct inoculation After transferring the contents of thecontainer or containers to be tested to the culture mediumadd an inoculum of a small number of viable micro-organ-isms (not more than 100 CFU) to the medium.
In both cases use the same micro-organisms as thosedescribed above under Growth promotion test. Perform agrowth promotion test as a positive control. Incubate all thecontainers containing medium for not more than 5 days. Ifclearly visible growth of micro-organisms is obtained afterthe incubation, visually comparable to that in the controlvessel without product, either the product possesses noantimicrobial activity under the conditions of the test orsuch activity has been satisfactorily eliminated. The test forsterility may then be carried out without further modiˆca-tion. If clearly visible growth is not obtained in the presenceof the product to be tested, visually comparable to that inthe control vessels without product, the product possessesantimicrobial activity that has not been satisfactorily elimi-nated under the conditions of the test. Modify the conditionsin order to eliminate the antimicrobial activity and repeat thevalidation test.
In the membrane ˆltration, the antimicrobial activity shouldbe suppressed by suitable means such as replacement of themembrane ˆlters with less adsorptive ones, increase of theamount of rinsing ‰uid, or addition of a suitable inactivatingagent to the rinsing ‰uid. Do not exceed a washing cycle of5 times 100 mL per ˆlter, even if during validation it hasbeen demonstrated that such a cycle does not fully eliminatethe antimicrobial activity.
In the direct inoculation, use a suitable inactivating agentwhich does not aŠect the growth of microorganisms or in-crease the volume of medium irrespective of the prescriptionin II-2 so that no antimicrobial activity remains.
Test for sterility of the products to be examinedNumber of articles to be tested
Items to be used for the test are taken from the lot accord-ing to an appropriate sampling plan prepared by referring tothe numbers speciˆed in Table 2.
Table 2. Number of items to be taken from the lot
Number of items in the lot Minimum number of items tobe tested for each medium*
InjectionsNot more than 100 containers
More than 100 but not morethan 500 containersMore than 500 containers
For large-volume products(More than 100 mL)
10z or 4 containers, which-ever is greater
10 containers
2z or 20 containers, which-ever is less2z or 10 containers, which-ever is less
Ophthalmic and othernon-injectable products
Not more than 200 containers
More than 200 containersIf the product is presented inthe form of single-dose con-tainers, apply the schemeshown above for preparationsfor parenteral use
5z or 2 containers, which-ever is greater10 containers
Bulk solid productsUp to 4 containersMore than 5 containers butnot more than 50 containersMore than 50 containers
Each container20z or 4 containers, which-ever is greater2z or 10 containers, which-ever is greater
*If the contents of one container are enough to inoculate thetwo media, this column gives the number of containers need-ed for both the media together.
Testing methodsThe test may be carried out using the technique of mem-
brane ˆltration or by direct inoculation of the culture mediawith the product to be examined. Appropriate negativecontrols are included. The technique of membrane ˆltrationis used whenever the nature of the product permits, that is,for ˆlterable aqueous preparations, for alcoholic or oilypreparations and for preparations miscible with or soluble inaqueous or oily solvents provided these solvents do not havean antimicrobial eŠect in the conditions of the test.
I. Membrane ˆltrationBy this method, a test article is ˆltered through a mem-
brane ˆlter, and the ˆlter is rinsed and incubated by beingtransferred to a medium or by adding a medium to the ˆltra-tion apparatus. Use membrane ˆlter made from suitablematerial having a nominal pore size of 0.45 mm or smaller.Use a ˆlter funnel sterilizable by the moist heat method orother methods and free from any leakage or back ‰ow whenˆltration is performed with the membrane in place. Thetechnique described below assumes that membranes about50 mm in diameter will be used. If ˆlters of a diŠerentdiameter are used the volumes of the dilutions and the
16761676 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
washings should be adjusted accordingly.I-1. Preparation of sample solution
a) Liquid medicine: Use as it is, as the sample solution.b) Solid medicine: In the case of a solid medicine, to be
administered after dissolving or suspending, the samplesolution is prepared with the provided solvent, isotonicsodium chloride solution or water to give the concentra-tion of use.
c) Oils and oily solutions: Oils and oily solutions ofsu‹ciently low viscosity may be ˆltered without dilutionthrough a dry membrane. Viscous oils may be diluted asnecessary with a suitable sterile diluent such as isopropylmyristate shown not to have antimicrobial activity in theconditions of the test.
d) Ointments and creams: Ointments in a fatty base andemulsions of the water-in-oil type may be diluted by usingsterile isopropyl myristate that has previously beenˆltered through a sterilizing membrane ˆlter or by usingother solvents not aŠecting the growth of microorgan-isms. Heat the sample preparation, if necessary, to notmore than 409C. In exceptional cases it may be necessaryto heat to not more than 449C.I-2. Quantities of sample solution to be testedUse for each medium not less than quantity of the product
prescribed in Table 3, unless otherwise speciˆed. If thecontents of one container are insu‹cient to inoculate thetwo media, twice or more containers shown in Table 2 areused. When using the technique of membrane ˆltration, use,whenever possible, the whole contents of the container, butnot less than the quantities indicated in Table 3, dilutingwhere necessary to about 100 mL with a suitable sterilerinsing ‰uid.
Table 3. Minimum quantity to be used for each medium
Quantity per container Minimum quantity to beused for each medium
LiquidsLess than 1 mL
1 – 40 mL
Greater than 40 mL andnot greater than 100 mLGreater than 100 mL
Antibiotic liquidsOther preparations solublein water or in isopropylmyristate
The whole contents of eachcontainerHalf the contents of eachcontainer but not less than1 mL20 mL
10z of the contents of thecontainer but not less than20 mL1 mLThe whole contents of eachcontainer to provide not lessthan 200 mg
Insoluble preparations, creamsand ointments to be suspendedor emulsiˆed
Use the contents of eachcontainer to provide not lessthan 200 mg
SolidsLess than 50 mg
50 mg or more but less than300 mg300 mg – 5 gGreater than 5 g
The whole contents of eachcontainerHalf the contents of each con-tainer but not less than 50 mg150 mg500 mg
I-3. ProceduresUsually complete the ˆltration of the sample solution with
one or two separate ˆlter funnels. Transfer the contents ofthe container or containers to be tested to the membrane ormembranes. If the sample solution is not readily ˆlterable, itmay be further diluted with rinsing ‰uid and thereafterˆltered. Rinse the membrane(s) with each 100-mL of rinsing‰uid per ˆlter for established cycles in the validation test.Provided the sample does not have antimicrobial activity,the rinsing procedure can be omitted. Employ either of thetwo methods described below for incubation of themembrane(s). Use the same volume of each medium as in thevalidation test.
(1) The processed membrane is aseptically transferredfrom the apparatus and cut into two equal parts, or half thevolume of sample solution is ˆltered into an entire mem-brane. Transfer each half of the cut membrane, or eachwhole membrane into the medium.
(2) After ˆltration of sample solution into the apparatusto which the membrane ˆlters are ˆtted, each medium isadded to the apparatus itself.
II. Direct inoculation of the culture mediumThis is the method by which the entire content or a portion
of the content of a sample container is transferred directly tothe culture medium and incubated. Usually, this method isapplied for medicines to which the membrane ˆltrationmethod cannot be applied or for which the application of thedirect transfer method, rather than the membrane ˆltration
16771677Supplement II, JPXIV General Tests, Processes and Apparatus
method, is rational.For products containing a mercurial preservative that
cannot be tested by the membrane-ˆltration method, ‰uidthioglycolate medium incubated at 20 – 259C may be usedinstead of Soybean-casein digest medium.
II-1. Preparation of sample solutionUsually, proceed as directed for the membrane ˆltration
method. In the case of an insoluble medicine, the product issuspended or crushed in a suitable manner and used as asample.a) Oily liquids. Use media to which have been added a suita-
ble emulsifying agent at a concentration shown to beappropriate in the validation of the test, for examplepolysorbate 80 at a concentration of 10 g/L.
b) Ointments and creams. Prepare by diluting to about 1 in10 by emulsifying with the chosen emulsifying agent in asuitable sterile diluent such as a 1 g/L neutral solution ofmeat or casein peptone. Transfer the diluted product to amedium not containing an emulsifying agent.II-2. Quantities of sample solution to be testedTransfer the quantity of the preparation to be examined
prescribed in Table 3, by using pipette, syringe or othersuitable inoculation devices, directly into the culturemedium so that the volume of the product is not more than10z of the volume of the medium, unless otherwiseprescribed. Shake cultures containing oily products gentlyeach observation day. However when thioglycolate mediumis used for the detection of anaerobic microorganisms keepshaking or mixing to a minimum in order to maintainanaerobic conditions.
Cultivation and observationFluid thioglycolate medium and Alternative thioglycolate
medium are to be incubated at 30 – 359C and Soybean-casein digest medium is to be incubated at 20 – 259C for notless than 14 days. Observe the cultures several times duringthe incubation period. If the material being tested rendersthe medium turbid so that the presence or absence ofmicrobial growth cannot be readily determined by visualexamination, 14 days after the beginning of incubationtransfer suitable portions of the medium to fresh vessels ofthe same medium and then incubate the original and transfervessels for not less than 4 days.
Observation and interpretation of resultsIf no evidence of microbial growth is found, the product
to be examined complies with the test for sterility. Ifevidence of microbial growth is found the product examineddoes not comply with the test for sterility, unless it can beclearly demonstrated that the test was invalid for causesunrelated to the product to be examined. If no evidence ofmicrobial growth is found in the repeat test the productcomplies with the Sterility Test. If microbial growth is foundin the repeat test the product does not comply with theSterility Test.
70. Reference Standards;Reagents, Test Solutions; StandardSolutions for Volumetric Analysis;
Standard Solutions; MatchingFluids for Color; Optical Filters forWavelength and Transmission Rate
Cyclohexylamine for thin-layer chromatographyDicyclohexylurea for thin-layer chromatography
Change the following:
Calcium hydroxide for pH determination Calciumhydroxide prepared for pH determination.
[6]-Gingerol for thin-layer chromatography C17H26O4
A yellow-white to yellow, liquid or solid. Freely soluble inmethanol, in ethanol (99.5) and in diethyl ether, and practi-cally insoluble in water.
Purity Related substances—Dissolve 1.0 mg of [6]-gin-gerol for thin-layer chromatography in exactly 2 mL ofmethanol. Perform the test with 10 mL of this solution asdirected in the Identiˆcation under Ginger: any spot otherthan the principal spot at the Rf value of about 0.3 does notappear.
Piperidine hydrochloride C5H11N.HCl A white crys-talline powder. Dissolves in water and in methanol. The pHof the aqueous solution (1 in 20) is between 3.0 and 5.0.
Melting point: 247 – 2529CPurity Clarity and color of solution—Dissolve 1.0 g of
piperidine hydrochloride in 20 mL of water: the solution isclear and colorless.
Residue on ignition: not more than 0.10z (1 g).Content: not less than 99.0z. Assay—Dissolve about
0.25 g of piperidine hydrochloride, accurately weighed, in50 mL of water, add 5 mL of diluted nitric acid (1 in 3), andtitrate with 0.1 mol/L silver nitrate VS (potentiometric titra-tion). Perform a blank determination in the same manner,
16781678 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
and make any necessary correction.
Each mL of 0.1 mol/L silver nitrate VS＝12.16 mg of C5H11N.HCl
Sodium 1-decanesulfonate C10H21NaO3S A white pow-der.
Purity Clarity and color of solution—Dissolve 1.0 g in20 mL of water: the solution is clear and colorless.
Loss on drying: not more than 3.0z (1 g, 1059C,3 hours).
Content: not less than 98.0z. Assay—Weigh accuratelyabout 0.45 g of sodium 1-decanesulfonate, dissolve in 50 mLof water, and pass through a column, about 1.2 cm in insidediameter and about 25 cm in length, packed with about20 mL of strongly acidic ion-exchange resin (0.3 to 1.0 mm,H type) at a ‰ow rate of about 4 mL per minute. Wash with150 mL of water at a ‰ow rate of about 4 mL per minute.Combine the washing and the elute, and titrate with0.1 mol/L sodium hydroxide VS (potentiometric titration).Perform a blank determination in the same manner, andmake any necessary correction.
Each mL of 0.1 mol/L sodium hydroxide VS＝24.43 mg of C10H21NaO3S
Thioglycolate medium I for sterility test See ‰uidthioglycolate medium.
Thioglycolate medium II for sterility test See alternativethioglycolate medium.
Turmeric paper Macerate 20 g of powdered driedrhizome of Curcuma longa Linn áe with four 100 mL-portionsof cold water, decant the supernatant liquid each time, anddiscard it. Dry the residue at a temperature not over 1009C.Macerate the dried residue with 100 mL of ethanol (95) forseveral days, and ˆlter. Immerse ˆlter paper in this ethanoldecoction, and allow the ethanol (95) to evaporate spontane-ously in clean air.
Sensitivity—Dip a strip of turmeric paper, about 1.5 cmlength, in a solution of 1 mg of boric acid in a mixture of1 mL of hydrochloric acid and 4 mL of water, after 1 minuteremove the paper from the liquid, and allow it to dry spon-taneouly: the yellow color changes to brown. When the stripis moistened with ammonia TS, the color of the stripchanges to greenish black.
L-Tyrosine C9H11NO3 White, crystals or crystallinepowder. Odorless and tasteless. Freely soluble in formicacid, very slightly soluble in water, and practically insolublein ethanol (95) and in diethyl ether. It dissolves in dilutehydrochloric acid and in dilute nitric acid.
Loss on drying: not more than 0.30z (1 g, 1059C,3 hours).
Content: not less than 99.0z. Assay—Weigh accuratelyabout 0.3 g of L-tyrosine, previously dried, dissolve in 6 mLof formic acid, add 50 mL of acetic acid (100), and titratewith 0.1 mol/L perchloric acid VS (potentiometric titra-tion). Perform a blank determination, and make any neces-
sary correction.
Each mL of 0.1 mol/L perchloric acid VS＝18.12 mg of C9H11NO3
Add the following:
0.05 mol/L Acetic acid-sodium acetate buŠer solution,pH 4.6 Dissolve 6.6 g of sodium acetate trihydrate in900 mL of water, and add 3 mL of acetic acid and water tomake 1000 mL.
0.25 mol/L Acetic acid TS To 3 g of acetic acid (100)add water to make 200 mL.
Aconitine for purity C34H47NO11 White, crystals or crys-talline powder. Sparingly soluble in acetonitrile and inethanol (99.5), slightly soluble in diethyl ether, and practi-cally insoluble in water. Melting point: about 1859C (withdecomposition).
Identiˆcation—Determine the infrared absorption spec-trum of aconitine for purity as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry: it exhibits absorption at the wave numbers of about3500 cm－1, 1718 cm－1, 1278 cm－1, 1111 cm－1, 1097 cm－1
and 717 cm－1.Absorbance E1z
1 cm (230 nm): 211 – 243 [5 mg dried fornot less than 12 hours in a desiccator (reduced pressure notexceeding 0.67 kPa, phosphorus (V) oxide, 409C), ethanol(99.5), 200 mL].
Purity Related substances—(1) Dissolve 5.0 mg of aconitine for purity in 2 mL of
acetonitrile, and use as the sample solution. Pipet 1 mL ofthe sample solution, add acetonitrile to make exactly 50 mL,and use as the standard solution. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 20 mL each of the sample solution and the standardsolution on a plate of silica gel for thin-layer chro-matography, and proceed the test as directed in the Identiˆ-cation in Processed Aconite Root: the spot other than theprincipal spot obtained with the sample solution is not moreintense than the spot with the standard solution.
(2) Dissolve 5.0 mg of aconitine for purity in 5 mL ofacetonitrile, and use as the sample solution. Pipet 1 mL ofthe sample solution, add acetonitrile to make exactly 50 mL,and use as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the totalarea of the peaks other than the peaks of aconitine and thesolvent obtained with the sample solution is not larger thanthe peak area of aconitine with the standard solution.Operating conditions
Detector, column, and column temperature: Proceed asdirected in the operating conditions in the Purity underProcessed Aconitine Root.
Mobile phase: A mixture of phosphate buŠer solution forprocessed aconite root and tetrahydrofuran (9:1).
Flow rate: Adjust the ‰ow rate so that the retention time
16791679Supplement II, JPXIV General Tests, Processes and Apparatus
of aconitine is about 26 minutes.Time span of measurement: About 3 times as long as the
retention time of aconitine.System suitability
Test for required detectability: Pipet 1 mL of the standardsolution, and add acetonitrile to make exactly 20 mL.Conˆrm that the peak area of aconitine obtained from 10 mLof this solution is equivalent to 3.5 to 6.5z of that obtainedfrom 10 mL of the standard solution.
System performance: Dissolve 1 mg each of aconitine forpurity, hypaconitine for purity and mesaconitine for purity,and 8 mg of jesaconitine for purity in 200 mL of acetonitrile.When the procedure is run with 10 mL of this solution underthe above operating conditions, mesaconitine, hypaconitine,aconitine and jesaconitine are eluted in this order, and eachresolution between these peaks is not less than 1.5, respec-tively.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of aconitine is not more than 1.5z.
Water: not more than 1.0z [5 mg dried for not less than12 hours in a desiccator (reduced pressure not exceeding0.67 kPa, phosphorus (V) oxide, 409C), coulometric titra-tion].
Aconitum diester alkaloids standard solution for purityIt is a solution containing 10 mg of aconitine for purity,10 mg of jesaconitine for purity, 30 mg of hypaconitine forpurity and 20 mg of mesaconitine for purity in 1000 mL of amixture of phosphate buŠer solution for processed aconiteroot and acetonitrile (1:1). When proceed the test with 20 mLof this solution as directed in the Purity under ProcessedAconite Root, using the detection wavelength, 231 nm, thepeaks of aconitine, jesaconitine, hypaconitine andmesaconitine are observed, and the ratio of their peakheights is about 10:1:35:30. When proceed the test using thedetection wavelength, 254 nm, the peaks of aconitine,jesaconitine, hypaconitine and mesaconitine are observed,and the ratio of their peak heights is about 2:8:7:6.
Alternative thioglycolate medium See the Sterility Testunder the General Tests, Processes and Apparatus.
4-(Aminomethyl)benzoic acid C8H9NO2 A white pow-der.
Purity—Dissolve 10 mg of 4-(aminomethyl)benzoic acidin 100 mL of water, and use this as the sample solution.Pipet 1 mL of the sample solution, add water to makeexactly 20 mL, and use this solution as the standardsolution. Perform the test with exactly 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the operatingconditions as directed in the Purity (5) under TranexamicAcid, and determine each peak area by the automaticintegration method: each area of the peak other than 4-(aminomethyl)benzoic acid obtained from the sample solu-tion is not more than the peak area of 4-(aminomethyl)ben-zoic acid from the standard solution.
Ammonium amminetrichloroplatinate for liquid chro-matography Cl3H7N2Pt To 20 g of cisplatin add 600 mLof 6 mol/L hydrochloric acid TS, and heat under a re‰uxcondenser for 4 – 6 hours to boil while stirring. After cool-ing, evaporate the solvent, and dry the orange residue atroom temperature under reduced pressure. To the residue soobtained add 300 mL of methanol, and heat at about 509Cto dissolve. Filter, separate insoluble yellow solids, and washthe solids with 10 mL of methanol. Combine the ˆltrate andthe washing, heat at about 509C, and add slowly 100 mL ofethyl acetate while stirring. Cool the mixture to roomtemperature avoiding exposure to light, and allow to standat －109C for 1 hour. Filter the mixture to take oŠ theformed crystals, wash the crystals with 100 mL of acetone,combine the washing to the ˆltrate, and evaporate to drynessto obtain orange crystals. If necessary, repeat the puriˆca-tion procedure described above to take oŠ the insolublecrystals. To the orange crystals obtained add 300 to 500 mLof a mixture of acetone and methanol (5:1), and heat atabout 509C while stirring to dissolve. Filter while hot to takeoŠ the insoluble crystals, wash the crystals with the mixture,and combine the ˆltrate and washing. Repeat the procedureseveral times, and evaporate to dryness. Suspense thecrystals so obtained in 50 mL of acetone, ˆlter, wash thecrystals with 20 mL of acetone, and dry the crystals at roomtemperature under reduced pressure. It is a yellow-browncrystalline powder.
Identiˆcation—Determine the infrared absorption spec-trum of the substance to be examined, previously dried at809C for 3 hours, as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry: it exhibitsabsorption at the wave numbers of about 3480 cm－1,3220 cm－1, 1622 cm－1, 1408 cm－1 and 1321 cm－1.
Related substances—Cisplatin Conduct this procedureusing light-resistant vessels. Dissolve 10 mg in N,N-dimethylformamide to make exactly 10 mL, and use thissolution as the sample solution. Separately, dissolve 10 mgof Cisplatin in N,N-dimethylformamide to make exactly50 mL. Pipet 5 mL of this solution, add N,N-dimethylfor-mamide to make exactly 20 mL, and use this solution as thestandard solution. Perform the test with exactly 40 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the peak area of cisplatin bythe automatic integration method: the peak area from thesample solution is not more than that from the standardsolution.Operating conditions
Proceed as directed in the operating conditions in theAssay under Cisplatin.System suitability
System performance: When the procedure is run with40 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of cisplatin are not less than 2500 andnot more than 2.0, respectively.
System repeatability: When the test is repeated 6 timeswith 40 mL of the standard solution under the above
16801680 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
operating conditions, the relative standard deviation of thepeak area of cisplatin is not more than 5.0z.
Benidipine hydrochloride for assay C28H31N3O6.HCl[Same as the monograph Benidipine Hydrochloride. Whendried, it contains not less than 99.5z of benidipinehydrochloride (C28H31N3O6.HCl)]
Benzoin C6H5CH(OH)COC6H5 White to pale yellow,crystals or powder.
Melting point: 132 – 1379C
N-Benzoyl-L-isoleucyl-L-glutamyl(g-OR)-glycyl-L-arginyl-p-nitroanilide hydrochloride An equal amount mixture oftwo components, R＝H and R＝CH3. A white powder.Slightly soluble in water.
Benzoylmesaconine hydrochloride for thin-layer chro-matography C31H43NO10.HCl.xH2O White, crystals orcrystalline powder. Soluble in water and in ethanol (99.5).Melting point: about 2509C (with decomposition).
Purity Related substances—Dissolve 1.0 mg of ben-zoylmesaconine hydrochloride for thin-layer chro-matography in exactly 10 mL of ethanol (99.5), and use thissolution as the sample solution. Perform the test with thissolution as directed under the Thin-layer Chromatography.Spot 10 mL of the sample solution on a plate of silica gel forthin-layer chromatography. Proceed the test as directed inthe Identiˆcation under Processed Aconite Root: no spotother than the principal spot at around Rf 0.4 appears.
Content: not less than 98z. Assay—Weigh accurately0.25 g of 4,4?-bis(diethylamino)benzophenone, dissolve in50 mL of acetic acid (100), and titrate with 0.1 mol/Lperchloric acid VS (potentiometric titration). Perform ablank titration in the same manner, and make any necessarycorrection.
Each mL of 0.1 mol/L perchloric acid VS＝16.22 mg of C21H28N2O
Borate-hydrochloric acid buŠer solution, pH 9.0 Dis-solve 19.0 g of sodium borate in 900 mL of water, adjust thepH to exactly 9.0 with 1 mol/L hydrochloric acid TS, andadd water to make 1000 mL.
Bovine activated blood coagulation factor X A proteinobtained from bovine plasma. It has an activity to decom-pose prothrombin speciˆcally and limitedly and producethrombin. It does not contain thrombin and plasmin. Itcontains not less than 500 Units per mg protein. One unitindicates an amount of the factor X which hydrolyzes1 mmol of N-benzoyl-L-isoleucyl-L-glutamyl(g-OR)-glycyl-L-arginyl-p-nitroanilide in 1 minute at 259C.
Calibration ball for particle density measurement
Calibration ball with a known volume prepared for measure-ment of particle density. The volume of the calibration ballmust be accurately determined to the nearest 0.001 cm3.
Caprylic acid CH3(CH2)6COOH A clear and colorlessoily liquid, having a slight unpleasant odor. Freely soluble inethanol (95) and in chloroform, and very slightly soluble inwater.
Refractive index: nD20 1.426 – 1.430
Speciˆc gravity d420: 0.908 – 0.912
Distilling range 238 – 2429C, not less than 95 volz.
Chlorobutanol C4H7Cl3O [Same as the namesakemonograph in Part II]
Chlorogenic acid for thin-layer chromatographyC16H18O9.xH2O A white powder. Freely soluble inmethanol and in ethanol (99.5), and sparingly soluble inwater. Melting point: about 2059C (with decomposition).
Purity Related substances—Dissolve 1.0 mg of chloro-genic acid for thin-layer chromatography in 2 mL ofmethanol, and use this solution as the sample solution.Perform the test with this solution as directed under theThin-layer Chromatography. Spot 10 mL of the samplesolution on a plate of silica gel for thin-layer chro-matography, develop the plate with a mixture of ethylacetate, water and formic acid (6:1:1) to a distance of about10 cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 365 nm): no spot other than the principalspot at around Rf 0.5 appears.
Cisplatin [Same as the namesake monograph]
Cyclohexylamine C6H11NH2 A clear and colorlessliquid, having a characteristic amine-like odor. Miscible withwater, with N,N-dimethylformamide and with acetone.
Purity Related substances—Use cyclohexylamine as thesample solution. Separately, pipet 1 mL of cyclohexylamine,add hexane to make exactly 100 mL, and use this as the stan-dard solution. Perform the test as directed in the Thin-layerChromatography. Spot 5 mL each of the sample solution andthe standard solution on a plate of silica gel for thin-layerchromatography, develop the plate with a mixture of ethylacetate, methanol, ammonia water (28) and cyclohexane(6:2:1:1) to a distance of about 10 cm, and air-dry the plate.Allow the plate to stand in iodine vapor: the spot other thanthe principal spot obtained with the sample solution is notmore intense than the spot with the standard solution.
Cyclohexylmethanol C7H14O A liquid having slightcamphor odor. Soluble in ethanol (99.5).
Refractive index nD20: about 1.464
Bioling point: about 1859C
N,N?-Dibenzylethylenediamine diacetate A white toslightly pale yellow crystalline powder.
Identiˆcation—Determine the infrared absorption spec-trum of the substance to be examined as directed in thepotassium bromide disk method under the Infrared Spec-trophotometry: it exhibits absorption at the wave numbersof about 1530 cm－1, 1490 cm－1, 1460 cm－1, 1400 cm－1 and
16811681Supplement II, JPXIV General Tests, Processes and Apparatus
1290 cm－1.Content: not less than 99.0z. Assay—Weigh accurately
about 25 mg of N,N?-dibenzylethylenediamine diacetate,dissolve in 25 mL of methanol, and add a solution contain-ing 1.02 g of disodium hydrogen phosphate and 6.80 g ofpotassium dihydrogen phosphate in 1000 mL of water tomake exactly 50 mL. Pipet 5 mL of this solution, add amixture of the solution containing 1.02 g of disodium hydro-gen phosphate and 6.80 g of potassium dihydrogen phos-phate in 1000 mL of water and methanol (1:1) to makeexactly 20 mL, and use this solution as the sample solution.Separately, weigh accurately about 8 mg of acetic acid (100),add 25 mL of methanol, and add the solution containing1.02 g of disodium hydrogen phosphate and 6.80 g ofpotassium dihydrogen phosphate in 1000 mL of water tomake exactly 50 mL. Pipet 5 mL of this solution, add amixture of the solution containing 1.02 g of disodium hydro-gen phosphate and 6.80 g of potassium dihydrogen phos-phate in 1000 mL of water and methanol (1:1) to makeexactly 20 mL, and use this solution as the control solution.Perform the test with exactly 20 mL each of the sample solu-tion and the control solution as directed under the LiquidChromatography according to the following conditions, anddetermine each peak area by the automatic integrationmethod. After making correction for the peak areas basedon the valiance of the base-line and the peak of acetic acidon the chromatogram obtained with the sample solution,calculate the amount of N,N?-dibenzylethylenediamine bythe area percentage method.Operating conditions
Detector: An ultraviolet absorption photometer(wavelength: 220 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: A mixture of water, methanol and 0.25mol/L potassium dihydrogen phosphate TS, pH 3.5(11:7:2).
Flow rate: Adjust the ‰ow rate so that the retention timeof N,N?-dibenzylethylenediamine is about 4 minutes.
Time span of measurement: About 5 times as long as theretention time of N,N?-dibenzylethylenediamine.System suitability
System performance: Dissolve an amount of Benzyl-penicillin Benzathine, equivalent to about 85,000 Units, in25 mL of methanol, add a solution containing 1.02 g ofdisodium hydrogen phosphate and 6.80 g of potassiumdihydrogen phosphate in 1000 mL of water to make exactly50 mL. Pipet 5 mL of this solution, add a mixture of thesolution containing 1.02 g of disodium hydrogen phosphateand 6.80 g of potassium dihydrogen phosphate in 1000 mLof water and methanol (1:1) to make exactly 20 mL. Whenthe procedure is run with 20 mL of this solution under theabove operating conditions, N,N?-dibenzylethylenediamineand benzylpenicillin are eluted in this order with the resolu-
tion between these peaks being not less than 20.System repeatability: When the test is repeated 6 times
with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of N,N?-dibenzylethylenediamine is not more than2.0z.
1,2-Dichlorobenzene C6H4Cl2 A colorless liquid.Speciˆc gravity d4
20: 1.306Boiling point: 180 – 1819C
2,6-Dichlorophenol C6H4Cl2O White to purplish whitecrystals.
Melting point: 65 – 679C
Dicyclohexyl C12H22
Speciˆc gravity d2020: about 0.864
Boiling point: about 2279CMelting point: about 49C
Dicyclohexylurea C6H11NHCONHC6H11 A white crys-talline powder, having no odor.
Purity Related substances—Dissolve 50 mg of dicyclo-hexylurea in methanol to make 100 mL. Pipet 10 mL of thissolution, and add methanol to make 100 mL. Pipet 20 mL ofthis solution, add 10 mL of 0.5 mol/L sodium hydroxideTS, shake, then add 5 mL of diluted hydrochloric acid (1 in10), and shake. Perform the test with 50 mL of this solutionas directed under the Liquid Chromatography according tothe following conditions, determine the area of each peak bythe automatic integration method, and calculate the amountby the area percentage method: the total amount of thepeaks other than dicyclohexylurea is not more than 3.0z.Operating conditions
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Purity (5) (ii) under Acetohexamide.
Time span of measurement: About 5 times as long as theretention time of dicyclohexylurea after the solvent peak.System suitability
Test for required detectability: To exactly 5 mL of thestandard solution add water to make exactly 200 mL.Conˆrm that the peak area of dicyclohexylurea obtainedwith 50 mL of this solution is equivalent to 1.8 to 3.3z ofthat with 50 mL of the standard solution.
System performance, and system repeatability: Proceed asdirected in the system suitability in the Purity (5) (ii) underAcetohexamide.
Digoxin C41H64O14 [Same as the namesake mono-graph]
0.04 mol/L Disodium dihydrogen ethylenediamine tetraa-cetate TS Dissolve 14.890 g of disodium dihydrogenethylenediamine tetraacetate dihydrate in water to make1000 mL.
Ethylbenzene C6H5C2H5 A colorless liquid. Freelysoluble in acetone and in ethanol (99.5), and practically in-soluble in water.
Speciˆc gravity d420: 0.862¿0.872
16821682 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
Boiling point: about 1359C
Flopropione [Same as the namesake monograph]
Flopropione for assay [Same as the monographFlopropione. It contains not less than 99.0z of ‰opropione(C9H10O4), calculated on the anhydrous basis.]
Fluid thioglycolate medium See the Sterility Test underthe General Tests, Processes and Apparatus.
85z Glycerin C3H8O3 [Same as the monographGlycerin]
Human antithrombin III Serine protease inhibitionfactor obtained from normal plasma of health human. It is aprotein, which inhibits the activities of thrombin andactivated blood coagulation factor X. It contains not lessthan 300 Units per mg protein. One unit indicates an amountof the antithrombin III which inhibits 1 unit of thrombin at259C under the existence of heparin.
Dried human normal plasma powder Freeze-driednormal plasma obtained from healthy human.
Hydrogen peroxide-sodium hydroxide TS To a mixtureof water and hydrogen peroxide (30) (9:1) add 3 drops ofbromophenol blue TS, and then add 0.01 mol/L sodiumhydroxide TS until a purple-blue color develops. Preparebefore use.
N-(2-Hydroxyethyl)isonicotinamide nitric esterC8H9N3O4 A white crystalline powder.
Identiˆcation—Determine the infrared absorption spec-trum of N-(2-hydroxyethyl)isonicotinamide nitric ester asdirected in the potassium bromide disk method under theInfrared Spectrophotometry: it exhibits absorption at thewave numbers of about 3270 cm－1, 1653 cm－1, 1546 cm－1
and 1283 cm－1.
Hypaconitine for purity C33H45NO10 White, crystals orcrystalline powder. Soluble in acetonitrile, sparingly solublein ethanol (99.5) and in diethyl ether, and practically insolu-ble in water. Melting point: about 1759C (with decomposi-tion).
Identiˆcation—Determine the infrared absorption spec-trum of hypaconitine for purity as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry: it exhibits absorption at the wave numbers of about3500 cm－1, 1728 cm－1, 1712 cm－1, 1278 cm－1, 1118 cm－1,1099 cm－1 and 714 cm－1.
Absorbance E1z1 cm (230 nm): 217 – 252 [5 mg dried for
not less than 12 hours in a desiccator (reduced pressure notexceeding 0.67 kPa, phosphorus (V) oxide, 409C), ethanol(99.5), 200 mL].
Purity Related substances—(1) Dissolve 5.0 mg ofhypaconitine for purity in 2 mL of acetonitrile, and use asthe sample solution. Pipet 1 mL of the sample solution, addacetonitrile to make exactly 50 mL, and use as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 20 mL each ofthe sample solution and the standard solution on a plate of
silica gel for thin-layer chromatography, and proceed thetest as directed in the Identiˆcation in Processed AconiteRoot: the spot other than the principal spot obtained withthe sample solution is not more intense than the spot withthe standard solution.
(2) Dissolve 5.0 mg of hypaconitine for purity in 5 mLof acetonitrile, and use as the sample solution. Pipet 1 mL ofthe sample solution, add acetonitrile to make exactly 50 mL,and use as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the total area ofthe peaks other than the peaks of hypaconitine and the sol-vent obtained with the sample solution is not larger than thepeak area of hypaconitine with the standard solution.Operating conditions
Detector, column, and column temperature: Proceed asdirected in the operating conditions in the Purity underProcessed Aconite Root.
Mobile phase: A mixture of phosphate buŠer solution forprocessed aconite root and tetrahydrofuran (9:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof hypaconitine is about 23 minutes.
Time span of measurement: About 3 times as long as theretention time of hypaconitine.System suitability
Test for required detectability: Pipet 1 mL of the standardsolution, and add acetonitrile to make exactly 20 mL.Conˆrm that the peak area of hypaconitine obtained from10 mL of this solution is equivalent to 3.5 to 6.5z of thatobtained from 10 mL of the standard solution.
System performance: Dissolve 1 mg each of aconitine forpurity, hypaconitine for purity and mesaconitine for purity,and 8 mg of jesaconitine for purity in 200 mL of acetonitrile.When the procedure is run with 10 mL of this solution underthe above operating conditions, mesaconitine, hypaconitine,aconitine and jesaconitine are eluted in this order, and eachresolution between these peaks is not less than 1.5, respec-tively.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of hypaconitine is not more than 1.5z.
Water: not more than 1.0z [5 mg dried for not less than12 hours in a desiccator (reduced pressure not exceeding0.67 kPa, phosphorus (V) oxide, 409C), coulometric titra-tion].
Icariin for thin-layer chromatography C33H40O15 Lightyellow crystals. Very slightly soluble in methanol and inethanol (99.5), and practically insoluble in water. Meltingpoint: about 2349C (with decomposition).
Purity Related substances—Dissolve 1.0 mg of icariinfor thin-layer chromatography in 1 mL of methanol. Per-form the test with 10 mL of this solution as directed in theIdentiˆcation under Epimedium Herb: no spot other thanthe principal spot having Rf value about 0.4 appears.
16831683Supplement II, JPXIV General Tests, Processes and Apparatus
L-Isoleucine C6H13NO2 [Same as the namesake mono-graph]
Jesaconitine for purity C35H49NO12 A white powder.Freely soluble in acetonitrile, in ethanol (99.5) and in diethylether, and practically insoluble in water.
Identiˆcation—Determine the infrared absorption spec-trum of jesaconitine for purity as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry: it exhibits absorption at the wave numbers of about3500 cm－1, 1715 cm－1, 1607 cm－1, 1281 cm－1, 1259 cm－1,1099 cm－1 and 772 cm－1.
Absorbance E1z1 cm (258 nm): 270 – 291 [5 mg dried for
not less than 12 hours in a desiccator (reduced pressure notexceeding 0.67 kPa, phosphorus (V) oxide, 409C), ethanol(99.5), 200 mL].
Purity Related substances—(1) Dissolve 5.0 mg ofjesaconitine for purity in 2 mL of acetonitrile, and use as thesample solution. Pipet 1 mL of the sample solution, addacetonitrile to make exactly 50 mL, and use as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 20 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography, and proceed thetest as directed in the Identiˆcation in Processed AconiteRoot: the spot other than the principal spot is not more in-tense than the spot with the standard solution.
(2) Dissolve 5.0 mg of jesaconitine for purity in 5 mL ofacetonitrile, and use as the sample solution. Pipet 1 mL ofthe sample solution, add acetonitrile to make exactly 50 mL,and use as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the total area ofthe peaks other than the peaks of jesaconitine and the sol-vent is not larger than the peak area of jesaconitine with thestandard solution.Operating conditions
Detector, column, and column temperature: Proceed asdirected in the operating conditions in the Purity underProcessed Aconite Root.
Mobile phase: A mixture of phosphate buŠer solution forprocessed aconite root and tetrahydrofuran (9:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof jesaconitine is about 36 minutes.
Time span of measurement: About 3 times as long as theretention time of jesaconitine.System suitability
Test for required detectability: Pipet 1 mL of the standardsolution, and add acetonitrile to make exactly 20 mL.Conˆrm that the peak area of jesaconitine obtained from10 mL of this solution is equivalent to 3.5 to 6.5z of thatobtained from 10 mL of the standard solution.
System performance: Dissolve 5 mg each of aconitine forpurity, hypaconitine for purity and mesaconitine for purity,and 1 mg of jesaconitine for purity in 200 mL of acetonitrile.When the procedure is run with 10 mL of this solution under
the above operating conditions, mesaconitine, hypaconitine,aconitine and jesaconitine are eluted in this order, and eachresolution between these peaks is not less than 1.5, respec-tively.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of jesaconitine is not more than 1.5z.
Water: not more than 1.0z [5 mg dried for not less than12 hours in a desiccator (reduced pressure not exceeding0.67 kPa, phosphorus (V) oxide, 409C), coulometrictitration].
Loganin for thin-layer chromatography C17H26O10
White, crystals or crystalline powder. Soluble in water,sparingly soluble in methanol, and very slightly soluble inethanol (99.5). Melting point: 221 – 2279C.
Purity Related substances—Dissolve 1.0 mg of loganinfor thin-layer chromatography in 2 mL of methanol. Per-form the test with 10 mL of this solution as directed in theIdentiˆcation under Cornus Fruit: any spot other than theprincipal spot at the Rf value of about 0.4 does not appear.
Mesaconitine for purity C33H45NO11 White, crystals orcrystalline powder. Slightly soluble in acetonitrile and inethanol (99.5), very slightly soluble in diethyl ether, andpractically insoluble in water. Melting point: about 1909C(with decomposition).
Identiˆcation—Determine the infrared absorption spec-trum of mesaconitine for purity as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry: it exhibits absorption at the wave numbers of about3510 cm－1, 1713 cm－1, 1277 cm－1, 1116 cm－1, 1098 cm－1
and 717 cm－1.Absorbance E1z
1 cm (230 nm): 211 – 247 [5 mg dried fornot less than 12 hours in a desiccator (reduced pressure notexceeding 0.67 kPa, phosphorus (V) oxide, 409C), ethanol(99.5), 200 mL].
Purity Related substances—(1) Dissolve 5.0 mg ofmesaconitine for purity in 2 mL of acetonitrile, and use asthe sample solution. Pipet 1 mL of the sample solution, addacetonitrile to make exactly 50 mL, and use as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 20 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography, and proceed thetest as directed in the Identiˆcation under Processed AconiteRoot: the spot other than the principal spot is not more in-tense than the spot with the standard solution.
(2) Dissolve 5.0 mg of mesaconitine for purity in 5 mLof acetonitrile, and use as the sample solution. Pipet 1 mL ofthe sample solution, add acetonitrile to make exactly 50 mL,and use as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the total area ofthe peaks other than the peaks of mesaconitine and the sol-vent is not larger than the peak area of mesaconitine with the
16841684 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
standard solution.Operating conditions
Detector, column, and column temperature: Proceed asdirected in the operating conditions in the Purity underProcessed Aconite Root.
Mobile phase: A mixture of phosphate buŠer solution forprocessed aconite root and tetrahydrofuran (9:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof mesaconitine is about 19 minutes.
Time span of measurement: About 3 times as long as theretention time of mesaconitine.System suitability
Test for required detectability: Pipet 1 mL of the standardsolution, and add acetonitrile to make exactly 20 mL.Conˆrm that the peak area of mesaconitine obtained from10 mL of this solution is equivalent to 3.5 to 6.5z of thatobtained from 10 mL of the standard solution.
System performance: Dissolve 1 mg each of aconitine forpurity, hypaconitine for purity and mesaconitine for purity,and 8 mg of jesaconitine for purity in 200 mL of acetonitrile.When the procedure is run with 10 mL of this solution underthe above operating conditions, mesaconitine, hypaconitine,aconitine and jesaconitine are eluted in this order, and eachresolution between these peaks is not less than 1.5, respec-tively.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of mesaconitine is not more than 1.5z.
Water: not more than 1.0z [5 mg dried for not less than12 hours in a desiccator (reduced pressure not exceeding0.67 kPa, phosphorus (V) oxide, 409C), coulometric titra-tion].
Methanol for liquid chromatography CH3OH A clear,colorless liquid. Mixable with water.
Purity Ultraviolet-absorbing substances—Perform thetest as directed in the Ultraviolet-visible Spectrophotometryusing water as the blank: the absorbances at 210 nm, at220 nm, at 230 nm, at 240 nm and at 254 nm are not morethan 0.70, 0.30, 0.15, 0.07 and 0.02, respectively.
Metoclopramide for assay [Same as the monographMetoclopramide. When dried, it contains not less than99.0z of metoclopramide (C14H22ClN3O2).]
1,3-Naphthalenediol C10H8O2 Red-brown crystals orgray-brown powder. Freely soluble in water, in methanoland in ethanol (99.5). Melting point: about 1249C.
1,3-Naphthalenediol TS Dissolve 50 mg of 1,3-naphthalenediol in 25 mL of ethanol (99.5), and add 2.5 mLof phosphoric acid.
3-Nitrophenol C6H5NO3 A light yellow crystallinepowder.
Melting point: 96 – 999C
Osthole for thin-layer chromatography C15H16O3 Awhite crystalline powder, having no odor. Freely soluble inmethanol and in ethyl acetate, soluble in ethanol (99.5), and
practically insoluble in water. Melting point: 83 – 849C.Purity Related substances—Dissolve 1.0 mg of osthole
for thin-layer chromatography in 1 mL of methanol. Per-form the test with 10 mL of this solution as directed in theIdentiˆcation under Cnidium Monnieri Fruit: on spot ap-pears other than the principal spot at around Rf 0.3.
Oxytocin C43H66N12O12S2 [Same as the namesakemonograph]
D-Phenylglycine C8H9NO2 White, crystals or crystal-line powder. Slightly soluble in water.
Loss on drying: not more than 0.5z (1 g, 1059C,3 hours).
Content: not less than 98.5z. Assay—Weigh accuratelyabout 0.3 g of D-phenylglycine, previously dried, dissolve in3 mL of formic acid, add 50 mL of acetic acid (100), andtitrate with 0.1 mol/L perchloric acid VS (potentiometrictitration). Perform a blank determination in the samemanner, and make any necessary correction.
Each mL of 0.1 mol/L perchloric acid VS＝15.12 mg of C8H9NO2
Phenylsilanized silica gel for liquid chromatographyPrepared for liquid chromatography.
0.05 mol/L Phosphate buŠer solution, pH 3.5 To 1000mL of 0.05 mol/L potassium dihydrogen phosphate TS adda suitable amount of a solution of phosphoric acid (49 in10,000) to make a solution having pH 3.5.
Phosphate buŠer solution for processed aconite rootDissolve 19.3 g of disodium hydrogen phosphate 12-water in3660 mL of water, and add 12.7 g of phosphoric acid.
[6]-Shogaol for thin-layer chromatography C17H24O3
A pale yellow oil. Miscible with methanol, ethanol (99.5)and with diethyl ether, and practically insoluble in water.
Purity Related substances—Dissolve 1.0 mg of [6]-shogaol for thin-layer chromatography in 2 mL ofmethanol, and perform the test with this solution as directedunder the Thin-layer chromatography. Spot 10 mL on a plateof silica gel for thin-layer chromatography, develop the platewith a mixture of ethyl acetate and hexane (1:1) to a distanceof about 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate, heat at 1059Cfor 5 minutes, and allow to cool: no spot other than theprincipal spot at around Rf 0.5 appears.
0.0375 mol/L Sodium 1-decanesulfonate TS Dissolve3.665 g of sodium 1-decanesulfonate in 400 mL of water.
Sodium dihydrogen phosphate NaH2PO4 A white,powder or crystalline powder. Freely soluble in water, andvery slightly soluble in ethanol (99.5). It has a hygroscopicproperty.
A solution is acidic.
16851685Supplement II, JPXIV General Tests, Processes and Apparatus
Soybean-casein digest medium See the Sterility Testunder the General Tests, Processes and Apparatus.
Thioacetamide C2H5NS A white crystalline powder orcolorless crystals. Freely soluble in water and in ethanol(99.5). Melting point: 112 – 1159C
Thioacetamide TS To 0.2 mL of a solution of thioa-cetamide (1 in 25) add 1 mL of a mixture of 15 mL ofsodium hydroxide TS, 5 mL of water and 20 mL of 85zglycerin, and heat in a water bath for 20 seconds. Preparebefore use.
Tiaramide hydrochloride for assay [Same as themonograph Tiaramide Hydrochloride. When dried, it con-tains not less than 99.0z of tiaramide hydrochloride(C15H18ClN3O3S.HCl).]
Trichloroacetic acid TS for serrapeptase Dissolve 1.80 gof trichloroacetic acid and 1.80 g of anhydrous sodiumacetate in 5.5 mL of 6 mol/L acetic acid TS and water tomake 100 mL.
Trichloro‰uoromethane CCl3F A colorless liquid orgas.
Speciˆc gravity d 417.2: 1.494
Boiling point: 23.79C
Vasopressin C46H65N15O12S2 A white powder.Constituent amino acids—Perform the test as directed in
the Constituent amino acids under Oxytocin, and calculatethe respective molar ratios with respect to glycine: 0.9 – 1.1for aspartic acid, 0.9 – 1.1 for glutamic acid, 0.9 – 1.1 forproline, 0.8 – 1.1 for tyrosine, 0.9 – 1.1 for phenylalanine,0.9 – 1.1 for arginine and 0.8 – 1.1 for cystine, and not morethan 0.03 for other amino acids.
0.04 mol/L Zinc chloride TS Dissolve 5.452 g of zincchloride in water to make 1000 mL.
(3) Standard Solutions forVolumetric Analysis
Change the following:
Iodine, 0.05 mol/L1000 mL of this solution contains 12.690 g of iodine
(I: 126.90).Preparation—Dissolve 13 g of iodine in 100 mL of a solu-
tion of potassium iodide (2 in 5), add 1 mL of dilutehydrochloric acid and water to make 1000 mL, and stan-dardize the solution as follows:
Standardization—Measure exactly 15 mL of the iodinesolution, and titrate with 0.1 mol/L sodium thiosulfate VS(Indicator method: starch TS; or potentiometric titration:platinum electrode). In the indicator method, when thesolution assumes a pale yellow color as the end point isapproached, add 3 mL of starch TS, and continue the titra-tion until the blue color disappears. Calculate the molarityfactor.
Note: Store protected from light. This solution, if storedfor a long period, should be restandardized before use.
Add the following:
Sodium Thiosulfate, 0.002 mol/L1000 mL of this solution contains 0.4964 g of sodium
thiosulfate VS with freshly boiled and cooled water to makeexactly 50 times the initial volume.
Add the following:
74. Powder Particle DensityDetermination
Powder Particle Density Determination is a method todetermine particle density of powdered pharmaceuticaldrugs or raw materials of drugs, and the gas displacementpycnometer is generally used. The powder density by thismethod is determined with an assumption that the volume ofthe gas displaced by the powder in a closed system is equal tothe volume of the powder. The bulk density at loose packingor the tapped density at tapping express the apparentdensities of the powder, since interparticulate void volumeof the powder is assumed to contribute a part of the volumeof the powder. On the contrary, the pycnometric particledensity expresses the powder density nearly equal to the crys-tal density, since the volume of the powder, that is deductedwith void volume of open pores accessible to gas, is counted.
Powder particle density is expressed in mass per unitvolume (kg/m3), and generally expressed in g/cm3.
ApparatusThe schematic diagram of particle density apparatus for
gas displacement pycnometric measurement is shown inFigure 1. The apparatus consists of a test cell in which thesample is placed, a reference cell and a manometer.
Generally, helium is used as the measurement gas. Theapparatus has to be equipped with a system capable ofpressuring the test cell to the deˆned pressure through themanometer.
Calibration of apparatus The volumes of the test cell(Vc) and the reference cell (Vr) must be accurately deter-mined to the nearest 0.001 cm3, and to assure accuracy of theresults of volume obtained, calibration of the apparatus iscarried out as follows using a calibration ball of knownvolume for particle density measurement. The ˆnal pressures(Pf) are determined for the initial empty test cell followed bythe test cell placed with the calibration ball for particledensity measurement in accordance with the procedures, andVc and Vr are calculated using the equation described in thesection of Procedure. Calculation can be made taking intoaccount that the sample volume (Vs) is zero in the ˆrst run.
16861686 Supplement II, JPXIVGeneral Tests, Processes and Apparatus
Figure 1. Schematic diagram of a gas displacement py-cnometer
ProcedureThe measurement of the particle density is carried out
between 15 and 309C, and temperature must not vary bymore than 29C during the course of measurement.
Firstly, weigh the mass of the test cell and record it. Afterweighing out the amount of the sample as described in theindividual monograph and placing it in the test cell, seal thecell in the pycnometer. Secondly, introduce the measure-ment gas (helium) into the test cell, and remove volatilecontaminants in the powder. If necessary, keep the samplepowder under reduced pressure to remove the volatilecontaminants in advance and use it as the test sample formeasurement.
Open the valve which connects the reference cell with thetest cell, conˆrm with the manometer that the pressure inside
the system is stable, and then read the system referencepressure (Pr). Secondly, close the valve that connects to thetwo cells, and introduce the measurement gas into the testcell to achieve positive pressure. Conˆrm with the manome-ter that the pressure inside the system is stable, and then readthe initial pressure (Pi). Open the valve to connect the testcell with the reference cell. After conˆrming that the indica-tor of the manometer is stable, read the ˆnal pressure (Pf),and calculate the sample volume (Vs) with the followingequation.
Repeat the measurement sequence for the same powdersample until consecutive measurements of the samplevolume agree to within 0.5z, and calculate the mean ofsample volumes (Vs). Finally, unload the test cell, weigh themass of test cell, and calculate the ˆnal sample mass bydeducting the empty cell mass from the test cell mass. Thepowder particle density r is calculated by the followingequation.
r＝ mVs
r: Powder particle density (g/cm3)m: Final sample mass (g)Vs: Sample volume (cm3)
16871687
O‹cial Monographsfor Part I
Acetohexamideアセトヘキサミド
Change the origin/limits of content to read:
Acetohexamide, when dried, contains not less than98.0z and not more than 101.0z of C15H20N2O4S.
Change the Description to read:
Description Acetohexamide occurs as a white to yellowishwhite powder.
It is freely soluble in N,N-dimethylformamide, sparinglysoluble in acetone, slightly soluble in methanol and inethanol (99.5), and practically insoluble in water.
Melting point: about 1859C (with decomposition).
Change the Purity (5) to read:
Purity(5) Related substances (i) Cyclohexylamine—Dissolve
exactly 1.0 g of Acetohexamide in exactly 30 mL of 0.5mol/L sodium hydroxide TS, add exactly 5 mL of hexane,shake vigorously for 60 minutes, allow to stand for 5minutes, and use the upper layer as the sample solution.Separately, dissolve exactly 50 mg of cyclohexylamine in 0.5mol/L sodium hydroxide TS to make exactly 50 mL. Pipet 2mL of this solution, and add 0.5 mol/L sodium hydroxideTS to make exactly 300 mL. Pipet 30 mL of this solution,add exactly 5 mL of hexane, shake vigorously for 60minutes, allow to stand for 5 minutes, and use the upper lay-er as the standard solution. Perform the test with exactly 2mL each of the sample solution and the standard solution asdirected under the Gas Chromatography according to thefollowing conditions, and determine the peak area of cyclo-hexylamine by the automatic integration method: the peakarea of cyclohexylamine is not more than that with the stan-dard solution.Operating conditions—
Detector: A hydrogen ‰ame-ionization detector.Column: A fused-silica column 0.53 mm in inside di-
ameter and 30 m in length, coated the inner surface withmethylsilicone polymer for gas chromatography 1.5 mm inthickness.
Column temperature: A constant temperature of about909C.
Injection port temperature: A constant temperature ofabout 1509C.
Detector temperature: A constant temperature of about2109C.
Carrier gas: HeliumFlow rate: Adjust the ‰ow rate so that the retention time
of cyclohexylamine is about 4 minutes.Split ratio: 1:1
System suitability—System performance: When the procedure is run with 2 mL
of the standard solution under the above operatingconditions, the number of theoretical plates of the peak ofcyclohexylamine is not less than 8000.
System repeatability: When the test is repeated 6 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cyclohexylamine is not more than 5z.
(ii) Dicyclohexylurea—Dissolve exactly 1.0 g of Aceto-hexamide in exactly 10 mL of 0.5 mol/L sodium hydroxideTS, add exactly 20 mL of methanol, shake, then add exactly5 mL of diluted hydrochloric acid (1 in 10), shake vigorouslyfor 15 minutes, and centrifuge. Filter 10 mL or more of thesupernatant liquid through a membrane ˆlter with pore sizeof not larger than 0.5 mm. Discard the ˆrst 5 mL of theˆltrate, and use the subsequent ˆltrate as the sample solu-tion. Separately, dissolve exactly 50 mg of dicyclohexylureain methanol to make exactly 100 mL. Pipet 2 mL of this so-lution, and add methanol to make exactly 100 mL. Pipet 20mL of this solution, add exactly 10 mL of 0.5 mol/L sodiumhydroxide TS, shake, then add exactly 5 mL of dilutedhydrochloric acid (1 in 10), shake, and use this solution asthe standard solution. Perform the test with exactly 50 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the peak area of dicy-clohexylurea by the automatic integration method: the peakarea of dicyclohexylurea is not more than that with the stan-dard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 210 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: Dissolve 0.5 g of sodium hydroxide in 1000mL of 0.05 mol/L sodium dihydrogen phosphate TS, andadjust the pH to 6.5 with 0.5 mol/L sodium hydroxide TS.To 500 mL of this solution add 500 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention time
16881688 Supplement II, JPXIVO‹cial Monographs for Part I
of dicyclohexylurea is about 10 minutes.System suitability—
System performance: When the procedure is run with50 mL of the standard solution under the above operatingconditions, the number of theoretical plates of the peak ofdicyclohexylurea is not less than 10,000.
System repeatability: When the test is repeated 6 timeswith 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of dicyclohexylurea is not more than 2.0z.
(iii) Other related substances—Dissolve 0.10 g of Aceto-hexamide in 10 mL of acetone, and use this solution as thesample solution. Pipet 1 mL of the sample solution, and addacetone to make exactly 20 mL. Pipet two 1-mL portions ofthis solution, add acetone to make exactly 10 mL and25 mL, respectively, and use these solutions as the standardsolution (1) and the standard solution (2). Perform the testwith these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solution,the standard solution (1) and the standard solution (2) on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture of ethylacetate, methanol, ammonia water (28) and cyclohexane(6:2:1:1) to a distance of about 10 cm, and air-dry the plate.Examine under ultraviolet light (main wavelength: 254 nm):the spots other than the principal spot from the samplesolution are not more intense than spot from the standardsolution (1), and the number of them which are more intensethan the spot from the standard solution (2) is not morethan 4.
Alprostadil, when dried, contains not less than97.0z and not more than 103.0z of C20H34O5.
Description Alprostadil occurs as white crystals or crystal-line powder.
It is freely soluble in ethanol (99.5) and in tetrahydrofu-ran, slightly soluble in acetonitrile, and practically insolublein water.
Identiˆcation (1) The absorption spectrum of a solutionof Alprostadil in ethanol (99.5) (1 in 100,000) determined asdirected under the Ultraviolet-visible Spectrophotometry
shows no absorption between 210 nm and 350 nm. Separate-ly, to 10 mL of this solution add 1 mL of potassium hydrox-ide-ethanol TS, allow to stand for 15 minutes, and determinethe absorption spectrum in the same way. Compare thespectrum so obtained with the Reference Spectrum or thespectrum of a solution of Alprostadil Reference Standardprepared in the same manner as the sample solution: bothspectra exhibit similar intensities of absorption at the samewavelengths.
(2) Determine the infrared absorption spectrum ofAlprostadil, previously dried, as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of previously dried Alprostadil ReferenceStandard: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
Purity Related substances—Dissolve 4 mg of Alprostadilin 2 mL of a mixture of acetonitrile for liquid chro-matography and water (9:1), and use this solution as thesample solution. Pipet 0.5 mL of the sample solution, andadd the mixture of acetonitrile for liquid chromatographyand water (9:1) to make exactly 10 mL. Pipet 2 mL of thissolution, add the mixture of acetonitrile for liquid chro-matography and water (9:1) to make exactly 10 mL, and usethis solution as the standard solution. Perform the test withexactly 5 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the area of thepeaks, having the relative retention time of about 0.70 and1.26 with respect to alprostadil, is not larger than 1/2 timesthe peak area of alprostadil with the standard solution, thearea of the peaks, having the relative retention time of about0.88 and 1.18 with respect to alprostadil, is not larger thanthe peak area of alprostadil with the standard solution, thearea of the peaks other than alprostadil and the peaks men-tioned above is not larger than 1/10 times the peak area ofalprostadil with the standard solution and the total area ofthe peaks other than alprostadil is not larger than 2 times thepeak area of alprostadil with the standard solution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 5 times as long as theretention time of alprostadil after the solvent peak.System suitability—
Test for required detectability: Measure exactly 2 mL ofthe standard solution, add the mixture of acetonitrile forliquid chromatography and water (9:1) to make exactly20 mL. Conˆrm that the peak area of alprostadil obtainedwith 5 mL of this solution is equivalent to 7 to 13z of thatwith 5 mL of the standard solution.
16891689Supplement II, JPXIV O‹cial Monographs for Part I
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of alprostadil is not more than 1.5z.
Loss on drying Not more than 1.0z (0.1 g, in vacuum,phosphorus (V) oxide, 4 hours).
Assay Weigh accurately about 5 mg each of Alprostadiland Alprostadil Reference Standard, previously dried,dissolve in exactly 5 mL of the internal standard solution,add a mixture of acetonitrile for liquid chromatography andwater (9:1) to make 50 mL, and use these solutions as thesample solution and the standard solution, respectively.Perform the test with 5 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area ofalprostadil to that of the internal standard.
Amount (mg) of C20H34O5＝WS×QT
QS
WS: Amount (mg) of Alprostadil Reference Standard
Internal standard solution—A solution of dimethyl phtha-late in the mixture of acetonitrile for liquid chromatographyand water (9:1) (1 in 10,000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 196 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 9.07 g of potassium dihydrogenphosphate in water to make 1000 mL. Adjust the pH to 6.3with a solution prepared by dissolving 9.46 g of disodiumhydrogen phosphate in water to make 1000 mL, and diluteto 10 times its volume with water. To 360 mL of this solutionadd 110 mL of acetonitrile for liquid chromatography and30 mL of methanol for liquid chromatography.
Flow rate: Adjust the ‰ow rate so that the retention timeof alprostadil is about 10 minutes.System suitability—
System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, alprostadil and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 9.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of alprostadil to that of the internal standard isnot more than 1.0z.
Containers and storage Containers—Tight containers.Storage—Light-resistant, and at a temperature not
exceeding 59C.
Atropine Sulfate Injection硫酸アトロピン注射液
Change the origin/limits of content to read:
Atropine Sulfate Injection is an aqueous solutionfor injection.
It contains not less than 93.0z and not more than107.0z of the labeled amount of atropine sulfate[(C17H23NO3)2.H2SO4.H2O: 694.83].
Change the Identiˆcation (2) to read:
Identiˆcation(2) Evaporate an exactly measured volume of Atropine
Sulfate Injection, equivalent to 5 mg of Atropine Sulfateaccording to the labeled amount, on a water bath to dryness.After cooling, dissolve the residue in 1 mL of ethanol (95),and use this solution as the sample solution. If insolublesubstance remains, crush it, allow to stand, and use thesupernatant liquid as the sample solution. Separately,dissolve 10 mg of Atropine Sulfate Reference Standard in2 mL of ethanol (95), and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 5 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a mixture of acetone, water and ammonia water (28)(90:7:3) to a distance of about 10 cm, and dry the plate at809C for 10 minutes. After cooling, spray evenly Dragen-dorŠ's TS for spraying on the plate: the spots obtained fromthe sample solution and the standard solution show anorange color and the same Rf value.
Add the following next to Identiˆcation:
Bacterial endotoxins Less than 75 EU/mg.
Actual volume It meets the requirements of Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirement of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirement of the Insoluble Particu-late Matter Test for Injections.
Sterility Perform the test: it meets the requirement of theSterility Test.
Change the Assay to read:
Assay To an exactly measured volume of Atropine SulfateInjection, equivalent to about 5 mg of atropine sulfate[(C17H23NO3)2.H2SO4.H2O], add exactly 3 mL of the inter-nal standard solution and water to make 50 mL, and use this
16901690 Supplement II, JPXIVO‹cial Monographs for Part I
solution as the sample solution. Separately, weigh accuratelyabout 25 mg of Atropine Sulfate Reference Standard,separately determine its loss on drying in the same condi-tions as for Atropine Sulfate, and dissolve in water to makeexactly 50 mL. Pipet 10 mL of this solution, add exactly 3mL of the internal standard solution and water to make 50mL, and use this solution as the standard solution. Performthe test with 20 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of atropine sulfate tothat of the internal standard.
Amount (mg) of atropine sulfate[(C17H23NO3)2.H2SO4.H2O]
＝WS×QT
QS×
15×1.027
WS: Amount (mg) of Atropine Reference Standard, calcu-lated based on the dried basis
Internal standard solution—A solution of etilefrinehydrochloride (1 in 1000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 210 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: To 0.4 g of sodium lauryl sulfate add500 mL of diluted phosphoric acid (1 in 1000) to dissolve,and adjust the pH to 3.0 with sodium hydroxide TS. To240 mL of this solution add 70 mL of tetrahydrofuran, andmix.
Flow rate: Adjust the ‰ow rate so that the retention timeof atropine sulfate is about 16 minutes.System suitability—
System performance: When the procedure is run with 20mL of the standard solution under the above operatingconditions, the internal standard and atropine sulfate areeluted in this order with the resolution between these peaksbeing not less than 3.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of atropine sulfate to that of the internalstandard is not more than 1.5z.
Azithromycin Hydrate is the derivative oferythromycin.
It contains not less than 945 mg (potency) and notmore than 1030 mg (potency) per mg, calculated on theanhydrous basis. The potency of Azithromycin Hy-drate is expressed as mass (potency) of azithromycin(C38H72N2O12: 748.98).
Description Azithromycin Hydrate occurs as a whitecrystalline powder.
It is freely soluble in methanol and in ethanol (99.5), andpractically insoluble in water.
Identiˆcation Determine the infrared absorption spectrumof Azithromycin Hydrate as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Azithromycin Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.
Purity (1) Heavy metals—Proceed with 1.0 g ofAzithromycin Hydrate according to Method 2, and performthe test. Prepare the control solution with 1.0 mL ofStandard Lead Solution (not more than 10 ppm).
(2) Related substances—Being speciˆed separately.(3) Residual solvent—Being speciˆed separately.
Water Not less than 4.0z and not more than 5.0z (0.4 g,volumetric titration, direct titration).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately an amount of Azithromycin
16911691Supplement II, JPXIV O‹cial Monographs for Part I
Hydrate and Azithromycin Reference Standard, equivalentto about 50 mg (potency), dissolve each in an adequateamount of a mixture of acetonitrile and water (3:2), addexactly 2 mL of the internal standard solution and themixture of acetonitrile and water (3:2) to make 50 mL, anduse these solutions as the sample solution and the standardsolution. Perform the test with 5 mL each of the samplesolution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and determine the ratios, QT and QS, of the peak areaof azithromycin to that of the internal standard.
Amount [mg (potency)] of azithromycin (C38H72N2O12)
＝WS×QR
QS×1000
WS: Amount [mg (potency)] of Azithromycin ReferenceStandard
Internal standard solution—A solution of 4,4?-bis(diethylamino)benzophenone in acetonitrile (3 in 4000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 215 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized polyvinyl alcohol gel polymer for liquid chro-matography (5 mm in particle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 6.97 g of dipotassium hydrogenphosphate in about 750 mL of water, adjust the pH to 11.0with potassium hydroxide TS, and add water to make 1000mL. To 400 mL of this solution add 600 mL of acetonitrilefor liquid chromatography.
Flow rate: Adjust the ‰ow rate so that the retention timeof azithromycin is about 10 minutes.System suitability—
System performance: When the procedure is run with 5 mLof the standard solution under the above operatingconditions, azithromycin and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of azithromycin to that of the internalstandard is not more than 1.0z.
Containers and storage Containers—Tight containers.
Benidipine Hydrochloride, when dried, contains notless than 99.0z and not more than 101.0z ofC28H31N3O6.HCl.
Description Benidipine Hydrochloride occurs as a yellowcrystalline powder.
It is very soluble in formic acid, soluble in methanol,sparingly soluble in ethanol (99.5), and practically insolublein water.
A solution of Benidipine Hydrochloride in methanol (1 in100) shows no optical rotation.
Melting point: about 2009C (with decomposition).
Identiˆcation (1) Determine the absorption spectrum ofa solution of Benidipine Hydrochloride in methanol (1 in100,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofBenidipine Hydrochloride, previously dried, as directed inthe potassium chloride disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(3) To 5 mL of a solution of Benidipine Hydrochloride(1 in 10) add 5 mL of ammonia TS, heat on a water bath for5 minutes, cool, and ˆlter. The ˆltrate, which is acidiˆedwith dilute nitric acid, responds to the Qualitative Tests (2)for chloride.
Purity (1) Heavy metals—Proceed with 1.0 g of Benidi-pine Hydrochloride according to Method 2, and perform thetest. Prepare the control solution with 2.0 mL of StandardLead Solution (not more than 20 ppm).
(2) Related substances—Dissolve 20 mg of BenidipineHydrochloride in 100 mL of a mixture of water andmethanol (1:1), and use this solution as the sample solution.Pipet 1 mL of the sample solution, add the mixture of waterand methanol (1:1) to make exactly 500 mL, and use thissolution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-
16921692 Supplement II, JPXIVO‹cial Monographs for Part I
lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the peak areas ofbisbenzylpiperidyl ester having the relative retention time ofabout 0.35 with respect to benidipine, dehydro derivativehaving the relative retention time of about 0.75 and otherrelated substances are not larger than 1/2 times the peakarea of benidipine with the standard solution, and the totalarea of the peaks other than benidipine is not larger than thepeak area of benidipine with the standard solution. For thiscalculation, use the peak areas of bisbenzylpiperidyl esterand dehydro derivative after multiplying by their responsefactors, 1.6, respectively.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 237 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 10 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (3 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of 0.05 mol/L potassium di-hydrogen phosphate TS, pH 3.0, methanol and tetra-hydrofuran (65:27:8).
Flow rate: Adjust the ‰ow rate so that the retention timeof benidipine is about 20 minutes.
Time span of measurement: About 2 times as long as theretention time of benidipine after the solvent peak.System suitability—
Test for required detectability: Measure exactly 5 mL ofthe standard solution, and add the mixture of water andmethanol (1:1) to make exactly 20 mL. Conˆrm that thepeak area of benidipine obtained with 10 mL of this solutionis equivalent to 18 to 32z of that with 10 mL of the standardsolution.
System performance: Dissolve 6 mg of BenidipineHydrochloride and 5 mg of benzoin in 200 mL of themixture of water and methanol (1:1). When the procedure isrun with 10 mL of this solution under the above operatingconditions, benzoin and benidipine are eluted in this orderwith the resolution between these peaks being not lessthan 8.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of benidipine is not more than 3.5z.
Loss on drying Not more than 0.5z (0.5 g, 1059C,2 hours).
Residue on ignition Not more than 0.10z (1 g).
Assay Weigh accurately about 0.7 g of BenidipineHydrochloride, previously dried, dissolve in 10 mL offormic acid, add 70 mL of acetic anhydride, and titrate with0.1 mol/L perchloric acid VS (potentiometric titration).Perform a blank determination in the same manner, andmake any necessary correction.
Each mL of 0.1 mol/L perchloric acid VS＝54.20 mg of C28H31N3O6.HCl
Containers and storage Containers—Tight containers.
Add the following:
Benidipine Hydrochloride Tablets塩酸ベニジピン錠
Benidipine Hydrochloride Tablets contain notless than 95.0z and not more than 105.0z of thelabeled amount of benidipine hydrochloride(C28H31N3O6.HCl: 542.02).
Method of preparation Prepare as directed under Tablets,with Benidipine Hydrochloride.
Identiˆcation Shake well a quantity of powdered Benidi-pine Hydrochloride Tablets, equivalent to 10 mg of Benidi-pine Hydrochloride according to the labeled amount, with100 mL of methanol, and centrifuge. To 10 mL of the super-natant liquid add methanol to make 100 mL, and use this so-lution as the sample solution. Determine the absorptionspectrum of the sample solution as directed under theUltraviolet-visible Spectrophotometry: it exhibits maximabetween 235 nm and 239 nm, and between 350 nm and 360nm.
Purity Dehydro derivative—Weigh accurately the mass ofnot less than 20 Benidipine Hydrochloride Tablets, andpowder in an agate mortar. To an amount of the powder,equivalent to 20 mg of Benidipine Hydrochloride, add about80 mL of a mixture of diluted phosphoric acid (1 in 500) andmethanol (1:1), shake well, and add the mixture of dilutedphosphoric acid (1 in 500) and methanol (1:1) to makeexactly 100 mL. Filter through a membrane ˆlter with poresize of 0.45 mm, and use the ˆltrate as the sample solution.Separately, dissolve 20 mg of benidipine hydrochloride forassay in the mixture of diluted phosphoric acid (1 in 500) andmethanol (1:1) to make exactly 100 mL. Pipet 1 mL of thissolution, add the mixture of diluted phosphoric acid (1 in500) and methanol (1:1) to make exactly 100 mL, and usethis solution as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the peakarea of dehydro derivative having the relative retention timeof about 0.75 with respect to benidipine is not larger than1/2 times the peak area of benidipine with the standard solu-tion. For this calculation, use the peak area of dehydroderivative after multiplying by the relative response factor,1.6.Operating conditions—
Perform as directed in the operating conditions in theAssay.
16931693Supplement II, JPXIV O‹cial Monographs for Part I
System suitability—Test for required detectability: Measure exactly 2 mL of
the standard solution, and add the mixture of diluted phos-phoric acid (1 in 500) and methanol (1:1) to make exactly20 mL. Conˆrm that the peak area of benidipine obtainedwith 10 mL of this solution is equivalent to 7 to 13z of thatwith 10 mL of the standard solution.
System performance: Dissolve 6 mg of benidipinehydrochloride and 5 mg of benzoin in 200 mL of a mixtureof water and methanol (1:1). When the procedure is run with10 mL of this solution under the above operating conditions,benzoin and benidipine are eluted in this order with theresolution between these peaks being not less than 8.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of benidipine is not more than 2.0z.
Content uniformity Perform the test as directed in theAssay, and determine the content: it meets the requirementsof the Content Uniformity Test.
Dissolution Perform the test with 1 tablet of BenidipineHydrochloride Tablets at 50 revolutions per minuteaccording to Method 2 under the Dissolution Test using thesinker, using 900 mL of the 1st ‰uid under the Disintegra-tion Test as the dissolution medium. Withdraw 20 mL ormore of the dissolution medium 30 minutes after starting thetest for a 2-mg and a 4-mg tablet or 45 minutes after startingthe test for a 8-mg tablet, and ˆlter through a membraneˆlter with pore size of not more than 0.45 mm. Discard theˆrst 10 mL of the ˆltrate, pipet the subsequent V mL, andadd the 1st ‰uid to make exactly V? mL so that each mLcontains about 2.2 mg of benidipine hydrochloride(C28H31N3O6.HCl) according to the labeled amount. Pipet5 mL of this solution, add exactly 5 mL of the mobile phase,and use this solution as the sample solution. Separately,weigh accurately about 22 mg of benidipine hydrochloridefor assay, previously dried at 1059C for 2 hours, anddissolve in the mobile phase to make exactly 100 mL. Pipet2 mL of this solution, and add the mobile phase to makeexactly 50 mL. Pipet 5 mL of this solution, and add themobile phase to make exactly 20 mL. Pipet 5 mL of thissolution, add exactly 5 mL of the 1st ‰uid, and use this solu-tion as the standard solution. Perform the test with exactly50 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine the peak areas, AT
and AS, of benidipine: the dissolution rates for 2-mg or 4-mgtablet in 30 minutes and for 8-mg tablet in 45 minutes are notless than 80z and not less than 85z, respectively.
Dissolution rate (z) with respect to the labeled amount ofbenidipine hydrochloride(C28H31N3O6.HCl)
＝WS×AT
AS×
V?V×
1C×9
WS: Amount (mg) of benidipine hydrochloride for assayC: Labeled amount (mg) of benidipine hydrochloride
(C28H31N3O6.HCl) in 1 tablet.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 237 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of 0.05 mol/L potassium di-hydrogen phosphate TS, pH 3.0 and acetonitrile (11:9).
Flow rate: Adjust the ‰ow rate so that the retention timeof benidipine is about 5 minutes.System suitability—
System performance: When the procedure is run with50 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of benidipine are not less than 3000 andnot more than 2.0, respectively.
System repeatability: When the test is repeated 6 timeswith 50 mL of the standard solution under the aboveoperating conditions, the relative standard deviation of thepeak area of benidipine is not more than 1.5z.
Assay Shake 1 tablet of Benidipine Hydrochloride Tabletswith 40 mL of a mixture of diluted phosphoric acid (1 in 500)and methanol (1:1) to disintegrate, add the mixture ofdiluted phosphoric acid (1 in 500) and methanol (1:1) tomake exactly V mL so that each mL contains 40 mg ofbenidipine hydrochloride (C28H31N3O6.HCl), and cen-trifuge. Pipet 20 mL of the supernatant liquid, add exactly10 mL of the internal standard solution and the mixture ofdiluted phosphoric acid (1 in 500) and methanol (1:1) tomake 50 mL, and use this solution as the sample solution.Separately, weigh accurately about 40 mg of benidipinehydrochloride for assay, previously dried at 1059C for 2hours, and dissolve in the mixture of diluted phosphoric acid(1 in 500) and methanol (1:1) to make exactly 100 mL. Pipet2 mL of this solution, add exactly 10 mL of the internal stan-dard solution and the mixture of diluted phosphoric acid (1in 500) and methanol (1:1) to make 50 mL, and use this solu-tion as the standard solution. Perform the test with 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of benidipine to that of the internal stan-dard. Perform the test described above with not less than 10tablets, and calculate the average of these results.
Amount (mg) of benidipine hydrochloride(C28H31N3O6.HCl)
＝WS×QT
QS×
V1000
WS: Amount (mg) of benidipine hydrochloride for assay
Internal standard solution—A solution of benzoin in amixture of water and methanol (1:1) (13 in 200,000).Operating conditions—
16941694 Supplement II, JPXIVO‹cial Monographs for Part I
Detector: An ultraviolet absorption photometer(wavelength: 237 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 10 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (3 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of 0.05 mol/L potassium di-hydrogen phosphate TS, pH 3.0, methanol and tetra-hydrofuran (65:27:8).
Flow rate: Adjust the ‰ow rate so that the retention timeof benidipine is about 20 minutes.System suitability—
System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and benidipine are elutedin this order with the resolution between these peaks beingnot less than 8.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of benidipine to that of the internal standard isnot more than 1.0z.
Containers and storage Containers—Well-closed contain-ers.
Benzylpenicillin Benzathineベンジルペニシリンベンザチン
Change the origin/limits of content to read:
Benzylpenicillin Benzathine is the N,N?-diben-zylethylenediamine salt of a penicillin compoundhaving antibacterial activity produced by the growthof Penicillium species.
It contains not less than 1152 Units and not morethan 1272 Units per mg, calculated on the anhydrousbasis. The potency of Benzylpenicillin Benzathine isexpressed as unit calculated from the amount ofbenzylpenicillin sodium (C16H17N2NaO4S: 356.37).1 Unit of Benzylpenicillin Benzathine is equivalent to0.6 mg of benzylpenicillin sodium (C16H17N2NaO4S). Itcontains not less than 24.0z and not more than27.0z of N,N?-dibenzylethylenediamine (C16H20N2:240.34), calculated on the anhydrous basis.
Change the Purity (3) to read:
Purity(3) Related substances—Dissolve 70 mg of Benzyl-
penicillin Benzathine in 25 mL of methanol, add a solutionprepared by dissolving 1.02 g of disodium hydrogen phos-phate and 6.80 g of potassium dihydrogen phosphate inwater to make 1000 mL to make 50 mL, and use thissolution as the sample solution. Pipet 1 mL of the sample
solution, add the mobile phase A to make exactly 100 mL,and use this solution as the standard solution. Perform thetest with exactly 20 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine each peak area by the automatic integrationmethod: the area of the peak having the relative retentiontime of about 2.4 with respect to benzylpenicillin is not morethan 2 times the total area of the peaks of benzylpenicillinand N,N?-dibenzylethylenediamine obtained from the stan-dard solution, and the area of the peak other than benzyl-penicillin, N,N?-dibenzylethylenediamine and the peak hav-ing the relative retention time of about 2.4 is not more thanthe total area of the peaks of benzylpenicillin andN,N?-dibenzylethylenediamine obtained from the standardsolution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 220 nm).
Column: A stainless steel column 4.0 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase A: A mixture of water, methanol and0.25 mol/L potassium dihydrogen phosphate TS, pH 3.5(6:3:1).
Mobile phase B: A mixture of methanol, water and0.25 mol/L potassium dihydrogen phosphate TS, pH 3.5(6:3:1).
Flowing of the mobile phase: Control the gradient bymixing the mobile phases A and B as directed in thefollowing table.
Time after injectionof sample (min)
Mobile phaseA (z)
Mobile phaseB (z)
0 – 10 75 2510 – 20 75 ª 0 25 ª 10020 – 55 0 100
Flow rate: 1.0 mL/minTime span of measurement: About 3 times as long as the
retention time of benzylpenicillin after the solvent peak.System suitability—
Test for required detectability: To exactly 1 mL of thestandard solution add the mobile phase A to make exactly20 mL. Conˆrm that the peak area of benzylpenicillinobtained from 20 mL of this solution is equivalent to 3.5 to6.5z of that from the standard solution.
System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, N,N?-dibenzylethylenediamine and benzyl-penicillin are eluted in this order with the resolution betweenthese peaks being not less than 25.
System repeatability: When the test is repeated 3 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak
16951695Supplement II, JPXIV O‹cial Monographs for Part I
areas of benzylpenicillin is not more than 2.0z.Change the Assay to read:
Assay (1) Benzylpenicillin—Weigh accurately an amountof Benzylpenicillin Benzathine, equivalent to about 85,000Units, dissolve in 25 mL of methanol, and add a solutioncontaining 1.02 g of disodium hydrogen phosphate and6.80 g of potassium dihydrogen phosphate in 1000 mL ofwater to make exactly 50 mL. Pipet 5 mL of this solution,add a mixture of the solution containing 1.02 g of disodiumhydrogen phosphate and 6.80 g of potassium dihydrogenphosphate in 1000 mL of water and methanol (1:1) to makeexactly 20 mL, and use this solution as the sample solution.Separately, weigh accurately an amount of BenzylpenicillinPotassium Reference Standard, equivalent to about 85,000Units, and about 25 mg of N,N?-dibenzylethylenediaminediacetate, dissolve in 25 mL of methanol, and add thesolution containing 1.02 g of disodium hydrogen phosphateand 6.80 g of potassium dihydrogen phosphate in 1000 mLof water to make exactly 50 mL. Pipet 5 mL of this solution,add a mixture of the solution containing 1.02 g of disodiumhydrogen phosphate and 6.80 g of potassium dihydrogenphosphate in 1000 mL of water and methanol (1:1) to makeexactly 20 mL, and use this solution as the standardsolution. Perform the test with exactly 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and determine the peak areas, AT ands AS, ofbenzylpenicillin.
Amount (unit) of benzylpenicillin sodium (C16H17N2NaO4S)
＝WS×AT
AS
WS: Amount (unit) of Benzylpenicillin Potassium Refer-ence Standard
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 220 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: A mixture of water, methanol and 0.25mol/L potassium dihydrogen phosphate TS, pH 3.5(11:7:2).
Flow rate: Adjust the ‰ow rate so that the retention timeof benzylpenicillin is about 18 minutes.System suitability—
System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, N,N?-dibenzylethylenediamine and benzyl-penicillin are eluted in this order with the resolution betweenthese peaks being not less than 20.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak
areas of N,N?-dibenzylethylenediamine and benzylpenicillinare not more than 2.0z, respectively.
(2) N,N?-Dibenzylethylenediamine—Determine theareas, AT and AS, of the peak corresponding to N,N?-diben-zylethylenediamine on the chromatograms obtained in (1)with the sample solution and the standard solution.
Amount (z) of N,N?-dibenzylethylenediamine (C16H20N2)
＝WS
WT×
AT
AS×100×0.667
WS: Amount (mg) of N,N?-dibenzylethylenediaminediacetate
WT: Amount (mg) of the sample0.667: Conversion factor for the molecular mass
of N,N?-dibenzylethylenediamine diacetate(C16H20N2･2CH3COOH) to that of N,N?-diben-zylethylenediamine (benzathine, C16H20N2)
Add the following:
Cefepime Dihydrochloride forInjection注射用塩酸セフェピム
Cefepime Dihydrochloride for Injection is a prepa-ration for injection, which is dissolved before use.
It contains not less than 95.0z and not morethan 110.0z of the labeled amount of cefepime(C19H24N6O5S2: 480.56).
Method of preparation Prepare as directed under Injec-tions, with Cefepime Dihydrochloride.
Description Cefepime Dihydrochloride for Injectionoccurs as a white to pale yellow powder.
Identiˆcation (1) Dissolve 40 mg of Cefepime Di-hydrochloride in 2 mL of water, add 1 mL of a solution ofhydroxylammonium chloride (1 in 10) and 2 mL of sodiumhydroxide TS, allow to stand for 5 minutes, then add 3 mLof 1 mol/L hydrochloric acid TS and 3 drops of iron (III)chloride TS: a red-brown color develops.
(2) Determine the absorption spectrum of a solution ofCefepime Dihydrochloride for Injection (1 in 12,500) asdirected under the Ultraviolet-visible Spectrophotometry: itexhibits maxima between 233 nm and 237 nm and between255 nm and 259 nm.
pH The pH of a solution obtained by dissolving an amountof Cefepime Dihydrochloride for Injection, equivalent to0.5 g (potency) of Cefepime Dihydrochloride according tothe labeled amount, in 5 mL of water is between 4.0 and 6.0.
Purity (1) Clarity and color of solution—Dissolve anamount of Cefepime Dihydrochloride for Injection, equiva-lent to 0.5 g (potency) of Cefepime Dihydrochloride accord-ing to the labeled amount, in 5 mL of water: the solution isclear and colorless or light yellow. The color is not darker
16961696 Supplement II, JPXIVO‹cial Monographs for Part I
than Matching Fluid I.(2) N-Methylpyrrolidine—Weigh accurately an amount
of Cefepime Dihydrochloride for Injection, equivalent toabout 0.2 g (potency) of Cefepime Dihydrochloride accord-ing to the labeled amount, dissolve in diluted nitric acid (2 in625) to make exactly 20 mL, and use this solution as thesample solution. Separately, transfer 30 mL of water into a100-mL volumetric ‰ask, weigh accurately the mass of the‰ask, add about 0.125 g of N-methylpyrrolidine, then weighaccurately the mass, and add water to make exactly 100 mL.Pipet 4 mL of this solution, add diluted nitric acid (2 in3125) to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 100 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the peak areas of N-methyl-pyrrolidine, AT and AS, by the automatic integration methodwithin 20 minutes after the sample solution is prepared.Calculate the amount of N-methylpyrrolidine per mg (poten-cy) of Cefepime Dihydrochloride for Injection by thefollowing formula: not more than 1.0z.
Amount (z) of N-methylpyrrolidine
＝WS×f
WT×
AT
AS×
1125
WS: Amount (mg) of N-methylpyrrolidineWT: Amount [mg (potency)] of cefepime in the sample
takenf: Purity (z) of N-methylpyrrolidine
Operating conditions—Proceed as directed in the operating conditions in the
Purity (3) under Cefepime Dihydrochloride.System suitability—
Proceed as directed in the system suitability in the Purity(3) under Cefepime Dihydrochloride.
Water Not more than 4.0z (Weigh accurately about50 mg of Cefepime Dihydrochloride for Injection, dissolvein exactly 2 mL of methanol for Karl Fischer method, andperform the test with exactly 0.5 mL of this solution. Coulo-metric titration).
Bacterial endotoxins Less than 0.06 EU/mg (potency).
Mass variation It meets the requirements of the MassVariation Test.
Foreign insoluble matter Perform the test according toMethod 2: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the InsolubleParticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay Weigh accurately the mass of the contents of notless than 10 Cefepime Dihydrochloride for Injection. Weigh
accurately an amount of the content, equivalent to about60 mg (potency) of Cefepime Dihydrochloride according tothe labeled amount, dissolve in the mobile phase to makeexactly 50 mL, and use this solution as the sample solution.Separately, weigh accurately an amount of CefepimeDihydrochloride Reference Standard, equivalent to about60 mg (potency), dissolve in the mobile phase to makeexactly 50 mL, and use this solution as the standardsolution. Proceed as directed in the Assay under CefepimeDihydrochloride.
Amount [mg (potency)] of cefepime (C19H24N6O5S2)
＝WS×AT
AS×1000
WS: Amount [mg (potency)] of Cefepime DihydrochlorideReference Standard
Containers and storage Containers—Hermetic containers.Storage—Light-resistant.
Purity (1) Clarity and color of solution—Dissolve 1.0 gof Cefuroxime Sodium in 10 mL of water: the solution isclear, and its absorbance at 450 nm is not more than 0.25.
16971697Supplement II, JPXIV O‹cial Monographs for Part I
Ciclosporinシクロスポリン
Change the Purity (3) to read:
Purity(3) Related substances—Use the sample solution ob-
tained in the Assay as the sample solution. Pipet 2 mL of thesample solution, add a mixture of water and acetonitrile(1:1) to make exactly 200 mL, and use this solution as thestandard solution. Perform the test with exactly 20 mL eachof the sample solution and the standard soluton as directedunder the Liquid Chromatography according to the follow-ing conditions. Determine each peak area of both solutionsby the automatic integration method: the area of the peakother than the ciclosporin is not more than 0.7 times of thepeak area of ciclosporin from the standard solution, and thetotal area of all peaks other than the ciclosporin is not morethan 1.5 times of the peak area of ciclosporin from the stan-dard solution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 2 times as long as theretention time of ciclosporin after the solvent peak.System suitability—
Test for required detection: To exactly 2 mL of the stan-dard solution add a mixture of water and acetonitrile (1:1) tomake exactly 20 mL. Conˆrm that the peak area of ciclospo-rin obtained from 20 mL of this solution is equivalent to 7 to13z of that of ciclosporin obtained from 20 mL of thestandard solution.
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the aboveoperating conditions, the relative standard deviation of thepeak area of ciclosporin is not more than 3.0z.
Change the Assay to read:
Assay Weigh accurately about 30 mg each of Ciclosporinand Ciclosporin Reference Standard, previously determinedthe loss on drying as the same manner as Ciclosporin, anddissolve each in a mixture of water and acetonitrile (1:1) tomake exactly 25 mL, and use these solutions as the samplesolution and the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the peakareas, AT and AS, of ciclosporin.
Amount (mg) of C62H111N11O12
＝WS×AS
AT
WS: Amount (mg) of Ciclosporin Reference Standard,
calculated on the dried basis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 210 nm).Column: A stainless steel column 4 mm in inside diameter
and 25 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).Connect the sample injection port and the column with astainless steel tube 0.3 mm in inside diameter and 1 m inlength.
Column temperature: A constant temperature of about809C (including the sample injection port and the connectingtube).
Mobile phase: A mixture of water, acetonitrile, tert-butylmethyl ether and phosphoric acid (520:430:50:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof ciclosporin is about 27 minutes.System suitability—
System performance: Dissolve 3 mg of Ciclosporin U in2.5 mL of a mixture of water and acetonitrile (1:1), and add2.5 mL of the standard solution. When the procedure is runwith 20 mL of this solution under the above operatingconditions, ciclosporin U and ciclosporin are eluted in thisorder with the resolution between these peaks being not lessthan 1.2.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of ciclosporin is not more than 1.0z.
Cisplatin, when dried, contains not less than 98.0zand not more than 102.0z of Cl2H6N2Pt.
Description Cisplatin occurs as a yellow crystalline pow-der.
It is sparingly soluble in N,N-dimethylformamide, slightlysoluble in water, and practically insoluble in ethanol (99.5).
Identiˆcation (1) To 5 mL of a solution of Cisplatin (1 in2000) add 2 to 3 drops of a solution of tin (II) chloridedihydrate (1 in 100): a brown precipitate is formed.
(2) Determine the absorption spectrum of a solution ofCisplatin in a solution of sodium chloride in 0.01 mol/Lhydrochloric acid TS (9 in 1000) (1 in 2000) as directed underthe Ultraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of a
16981698 Supplement II, JPXIVO‹cial Monographs for Part I
solution of Cisplatin Reference Standard prepared in thesame manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.
(3) Determine the infrared absorption spectrum ofCisplatin as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofCisplatin Reference Standard: both spectra exhibit similarintensities of absorption at the same wave numbers.
(4) A solution of Cisplatin (1 in 2000) responds to theQualitative Test (1) for chloride.
Purity Ammonium amminetrichloroplatinate—Conductthis procedure using light-resistant vessels. Dissolve 50 mgof Cisplatin in a solution of sodium chloride (9 in 1000) tomake exactly 100 mL, and use this solution as the samplesolution. Separately, dissolve 10 mg of ammonium am-minetrichloroplatinate for liquid chromatography, previous-ly dried at 809C for 3 hours, in the solution of sodiumchloride (9 in 1000) to make exactly 200 mL. Pipet 2 mL ofthis solution, add the solution of sodium chloride (9 in 1000)to make exactly 20 mL, and use this solution as the standardsolution. Perform the test with exactly 40 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and determine the peak area of ammoniumamminetrichloroplatinate by the automatic integrationmethod: the peak area from the sample solution is not morethan that from the standard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 209 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with silica gel forliquid chromatography having quaternary ammoniumgroups (10 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A solution of ammonium sulfate (1 in 800).Flow rate: Adjust the ‰ow rate so that the retention time
of ammonium amminetrichloroplatinate is about 8 minutes.System suitability—
System performance: When the procedure is run with40 mL of the standard solution under the above operatingconditions, the number of theoretical plates and thesymmetry factor of the peak of ammonium am-minetrichloroplatinate are not less than 1500 and not morethan 2.0, respectively.
System repeatability: When the test is repeated 6 timeswith 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of ammonium amminetrichloroplatinate is not morethan 3.0z.
Loss on drying Not more than 0.1z (1 g, 1059C, 4 hours).
Assay Conduct this procedure using light-resistant vessels.Weigh accurately about 25 mg each of Cisplatin andCisplatin Reference Standard, previously dried, dissolve in
N,N-dimethylformamide to make exactly 25 mL, and usethese solutions as the sample solution and the standardsolution. Perform the test with exactly 40 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and determine the peak areas, AT and AS, ofcisplatin by the automatic integration method.
Amount (mg) of Cl2H6N2Pt＝WS×AT
AS
WS: Amount (mg) of Cisplatin Reference Standard
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 310 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 25 cm in length, packed with aminopropyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of ethyl acetate, methanol, waterand N,N-dimethylformamide (25:16:5:5).
Flow rate: Adjust the ‰ow rate so that the retention timeof cisplatin is about 4 minutes.System suitability—
System performance: When the procedure is run with40 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of cisplatin are not less than 3000 andnot more than 2.0, respectively.
System repeatability: When the test is repeated 6 timeswith 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cisplatin is not more than 1.0z.
Containers and storage Containers—Tight containers.
Anhydrous Citric Acid無水クエン酸
Change the Identiˆcation to read:
Identiˆcation Determine the infrared absorption spectrumof Anhydrous Citric Acid, previously dried at 1059C for2 hours, as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.
16991699Supplement II, JPXIV O‹cial Monographs for Part I
Clarithromycinクラリスロマイシン
Change the origin/limits of content to read:
Clarithromycin is a derivative of erythromycin.It contains not less than 950 mg (potency) and not
more than 1050 mg (potency) per mg, calculated on theanhydrous basis. The potency of Clarithromycin isexpressed as mass (potency) of clarithromycin(C38H69NO13).
Change the Melting point to read:
Melting point 220 – 2279C
Change the Purity (2) and (3) to read:
Purity(2) Arsenic—Prepare the test solution with 1.0 g of
Clarithromycin according to Method 3, and perform the test(not more than 2 ppm).
(3) Related substances—Weigh accurately about 0.1 g ofClarithromycin, dissolve in the mobile phase to make exactly20 mL, and use this solution as the sample solution.Separately, weigh accurately about 10 mg of ClarithromycinReference Standard, dissolve in the mobile phase to makeexactly 20 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and determine the each peak area by the automatic integra-tion method: the amount of each related substance calculat-ed on the anhydrous basis is not more than 2.0z, and thetotal of them is not more than 5.0z. Exclude any peak withan area of less than 0.05z.
Amount (z) of each related substance calculated on theanhydrous basis
＝WS
WT×
AT
AS×100
Total amount (z) of the related substances calculated on theanhydrous basis
＝WS
WT×
SAT
AS×100
WS: Amount (mg) of Clarithromycin Reference StandardWT: Amount (mg) of the sample, calculated on the anhy-
drous basisAS: Peak area of clarithromycin obtained with the stan-
dard solutionAT: Peak area of each related substance obtained with the
sample solutionSAT: Total area of the peaks other than clarithromycin
obtained with the sample solutionOperating conditions—
Detector, column, column temperature, mobile phase,
and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 5 times as long as theretention time of the main peak after 2 minutes of sampleinjection.System suitability—
Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 10 mL,and use this solution as the solution for system suitabilitytest. Conˆrm that when the procedure is run with 10 mL ofthe solution for system suitability test, the peak area ofclarithromycin is equivalent to 14 – 26z of that obtainedfrom the standard solution.
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standarddeviation of the peak area of clarithromycin is not morethan 3.0z.
Colchicineコルヒチン
Change to read except the structural formulaand chemical name:
Colchicine contains not less than 97.0z and notmore than 102.0z of C22H25NO6, calculated on theanhydrous basis and corrected by the amount of ethylacetate.
Description Colchicine occurs as a yellowish whitepowder.
It is very soluble in methanol, freely soluble in N,N-dimethylformamide, in ethanol (95) and in acetic anhydride,and sparingly soluble in water.
It is colored by light.
Identiˆcation (1) Determine the absorption spectrum ofa solution of Colchicine in ethanol (95) (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.
(2) To 1 g of potassium bromide for infrared absorptionspectrum add 0.5 mL of a solution of Colchicine inmethanol (1 in 50), grind thoroughly, and dry in vacuum at809C for 1 hour. Determine the infrared absorption spec-trum of this powder as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.
Optical rotation [a]D20: －235 –－2509(0.1 g calculated onthe anhydrous basis and corrected by the amount of ethyl
17001700 Supplement II, JPXIVO‹cial Monographs for Part I
acetate, ethanol (95), 10 mL, 100 mm).
Purity (1) Colchiceine—Dissolve 0.10 g of Colchicine in10 mL of water, and to 5 mL of this solution add 2 drops ofiron (III) chloride TS: no deˆnite green color develops.
(2) Chloroform and ethyl acetate—Weigh accuratelyabout 0.60 g of Colchicine, dissolve in exactly 2 mL of theinternal standard solution, add N,N-dimethylformamide tomake 10 mL, and use this solution as the sample solution.Separately, weigh 0.30 g of chloroform using a 100-mLvolumetric ‰ask containing about 20 mL of N,N-dimethyl-formamide, and add N,N-dimethylformamide to make ex-actly 100 mL. Pipet 2 mL of this solution, add N,N-dimethylformamide to make exactly 200 mL, and use thissolution as the standard solution (1). Separately, weigh ac-curately about 1.8 g of ethyl acetate using a 100-mL volu-metric ‰ask containing about 20 mL of N,N-dimethylfor-mamide, and add N,N-dimethylformamide to make exactly100 mL. Pipet 2 mL of this solution, add exactly 2 mL of theinternal standard solution and N,N-dimethylformamide tomake 10 mL, and use this solution as the standard solution(2). Perform the test with 2 mL each of the sample solutionand the standard solutions (1) and (2) as directed under theGas Chromatography according to the following conditions:the peak area of chloroform is not more than that from thestandard solution (1). Determine the ratios of the peak areaof ethyl acetate to that of the internal standard, QT and QS,of the sample solution and the standard solution (2), and cal-culate the amount of ethyl acetate by the following formula:the amount of ethyl acetate is not more than 6.0z.
Amount (z) of ethyl acetate (C4H8O2)＝WS
WT×
QT
QS×2
WS: Amount (g) of ethyl acetateWT: Amount (g) of the sample
Internal standard solution—A solution of 1-propanol inN,N-dimethylformamide (3 in 200)Operating conditions—
Detector: A hydrogen ‰ame-ionization detectorColumn: A fused silica column 0.53 mm in inside di-
ameter and 30 m in length, coated inside surface withpolyethylene glycol 20 M for gas chromatography 1.0 mm inthickness.
Column temperature: 609C for 7 minutes, then up to1009C at a rate of 409C per minute if necessary, and hold at1009C for 10 minutes.
Injection port temperature: A constant temperature ofabout 1309C
Detector temperature: A constant temperature of about2009C
Carrier gas: HeliumFlow rate: Adjust the ‰ow rate so that the retention time
of ethyl acetate is about 3 minutes.Split ratio: 1:20
System suitability—Test for required detectability: Pipet 2 mL of the standard
solution (2), and add N,N-dimethylformamide to makeexactly 25 mL. Pipet 1 mL of this solution, and add N,N-
dimethylformamide to make exactly 50 mL. Conˆrm thatthe peak area of ethyl acetate obtained from 2 mL of thissolution is equivalent to 0.11 to 0.21z of that obtained from2 mL of the standard solution (2).
System performance: To 1 mL of chloroform add N,N-dimethylformamide to make 10 mL. To 1 mL of this solu-tion add 2 mL of ethyl acetate and N,N-dimethylformamideto make 100 mL. To 2 mL of this solution add 2 mL of theinternal standard solution and N,N-dimethylformamide tomake 10 mL. When the procedure is run with 2 mL of thissolution under the above operating conditions, ethyl acetate,chloroform and the internal standard are eluted in this orderwith the resolution between the peaks of chloroform and theinternal standard being not less than 2.0.
System repeatability: When the test is repeated 3 timeswith 2 mL of the standard solution (2) under the aboveoperating conditions, the relative standard deviation of theratio of the peak area of ethyl acetate to that of the internalstandard is not more than 3.0z.
(3) Related substances Dissolve 60 mg of Colchicine in100 mL of diluted methanol (1 in 2). Pipet 1 mL of thissolution, add diluted methanol (1 in 2) to make exactly 100mL, and use this solution as the sample solution. Performthe test with 20 mL of the sample solution as directed underthe Liquid Chromatography according to the followingconditions, and determine each peak area by the automaticintegration method. Calculate the total amount of the peaksother than colchicine by the area percentage method: notmore than 5.0z.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4.6 mm in inside di-ameter and 25 cm in length, packed with octylsilanized silicagel for liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: To 450 mL of 0.05 mol/L potassium di-hydrogen phosphate TS add methanol to make 1000 mL.Adjust the pH to 5.5 with diluted phosphoric acid (7 in 200).
Flow rate: Adjust the ‰ow rate so that the retention timeof colchicine is about 7 minutes.
Time span of measurement: About 2 times as long as theretention time of colchicine after the solvent peak.System suitability—
Test for required detectability: Pipet 1 mL of the samplesolution, and add diluted methanol (1 in 2) to make exactly50 mL. Conˆrm that the peak area of colchicine obtainedfrom 20 mL of this solution is equivalent to 1.4 to 2.6z ofthat obtained from 20 mL of the sample solution.
System performance: When the procedure is run with20 mL of the sample solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of colchicine are not less than 6000 andnot more than 1.5, respectively.
System repeatability: When the test is repeated 6 timeswith 20 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak area
17011701Supplement II, JPXIV O‹cial Monographs for Part I
of colchicine is not more than 2.0z.
Water Not more than 2.0z (0.5 g, volumetric titration,back titration).
Assay Weigh accurately about 0.4 g of Colchicine, dissolvein 25 mL of acetic anhydride, and titrate with 0.05 mol/Lperchloric acid VS (potentiometric titration). Perform ablank determination in the same manner, and make anynecessary correction.
Each mL of 0.05 mol/L perchloric acid VS＝19.97 mg of C22H25NO6
Containers and storage Containers—Tight containers.Storage—Light-resistant.
Cytarabineシタラビン
Change the origin/limits of content to read:
Cytarabine, when dried, contains not less than98.5z and not more than 101.0z of C9H13N3O5.
Change the Description to read:
Description Cytarabine occurs as white crystals or crystal-line powder.
It is freely soluble in water, soluble in acetic acid (100),and very slightly soluble in ethanol (99.5).
It dissolves in 0.1 mol/L hydrochloric acid TS.Melting point: about 2149C (with decomposition).
Change the Identiˆcation to read:
Identiˆcation (1) Determine the absorption spectrum ofa solution of Cytarabine in 0.1 mol/L hydrochloric acid TS(1 in 100,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofCytarabine as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.
Delete the Absorbance:
Change the Purity (4) and (5) to read:
Purity(4) Related substances—Dissolve 0.10 g of Cytarabine in
10 mL of water, and use this solution as the sample solution.Pipet 1 mL of the sample solution, add water to makeexactly 200 mL, and use this solution as the standardsolution (1). Pipet 10 mL of the standard solution (1), add
water to make exactly 25 mL and use this solution as thestandard solution (2). Perform the test with these solutionsas directed under the Thin-layer Chromatography. Spot10 mL each of the sample solution and the standard solutions(1) and (2) on a plate of silica gel with ‰uorescent indicatorfor thin-layer chromatography. Develop the plate with1-butanol saturated with water to a distance of about 12 cm,and air-dry the plate. Examine under ultraviolet light (mainwavelength: 254 nm): the spots other than the principal spotfrom the sample solution are not more intense than the spotfrom the standard solution (1), and the number of themwhich are more intense than the spot from the standard solu-tion (2) is not more than two. Spray evenly acidic potassiumpermanganate TS on the plate: any spot other than the prin-cipal spot does not appear.
Deferoxamine Mesilateメシル酸デフェロキサミン
Change the Purity (6) to read:
Purity(6) Related substances—Dissolve 50 mg of Deferoxa-
mine Mesilata in 50 mL of the mobile phase, and use thissolution as the sample solution. Pipet 3 mL of the sample so-lution, add the mobile phase to make exactly 50 mL, and usethis solution as the standard solution. Perform the test withexactly 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions. Determine each peakarea of both solutions by the automatic integration method:the total area of all peaks other than the peak of deferoxa-mine is not larger than the peak area of deferoxamine fromthe standard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 230 nm).
Column: A stainless steel column 4 mm in inside diameterand 20 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (10 mm in particle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 1.32 g of diammonium hydrogenphosphate, 0.37 g of disodium dihydrogen ethylenediaminetetraacetate dihydrate and 1.08 g of sodium 1-heptanesul-fonate in 950 mL of water, and adjust the pH of thissolution to 2.8 with phosphoric acid. To 800 mL of thissolution add 100 mL of 2-propanol.
Flow rate: Adjust the ‰ow rate so that the retention timeof deferoxamine is about 15 minutes.
Time span of measurement: About two times as long asthe retention time of deferoxamine after the solvent peak.System suitability—
Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 100 mL.Conˆrm that the peak area of deferoxamine obtained from
17021702 Supplement II, JPXIVO‹cial Monographs for Part I
20 mL of this solution is equivalent to 1.5 to 2.5z of that ofdeferoxamine obtained from 20 mL of the standard solution.
System performance: Dissolve 16 mg of DeferoxamineMesilate and 4 mg of methyl parahydroxybenzoate in 50 mLof the mobile phase. When the procedure is run with 20 mLof this solution under the above operating conditions,deferoxamine and methyl parahydroxybenzoate are eluted inthis order with the resolution between these peaks being notless than 4.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakareas of deferoxamine is not more than 3.0z.
Digoxinジゴキシン
Change the Description to read:
Description Digoxin occurs as colorless or white crystals ora white crystalline powder.
It is freely soluble in pyridine, slightly soluble in ethanol(95), very slightly soluble in acetic acid (100), and practicallyinsoluble in water.
Change the Identiˆcation (2) to read:
Identiˆcation(2) Determine the infrared absorption spectrum of
Digoxin, previously dried, as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.
Purity(2) Related substances—Dissolve exactly 25.0 mg of
Digoxin in 50 mL of warm ethanol (95), cool, and addethanol (95) to make exactly 100 mL. Pipet 10 mL of thissolution, add 10 mL of water and dilute ethanol to make 50mL, and use this solution as the sample solution. Separately,dissolve exactly 5.0 mg of Gitoxin Reference Standard,previously dried under reduced pressure at 1059C for 1 hour,in a mixture of acetonitrile and water (7:3) to make exactly200 mL. Pipet 2 mL of this solution, add dilute ethanol tomake 50 mL, and use this solution as the standard solution.Perform the test with exactly 10 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas, AT and AS, of gitoxin: AT is notlarger than AS, and the total of the areas of the peaks other
than digitoxin and gitoxin, obtained by the area percentagemethod, is not more than 3z.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 4 times as long as theretention time of digoxin after the solvent peak.System suitability—
Test for required detectability: Pipet 1 mL of the samplesolution, add the mobile phase to make exactly 100 mL, anduse this solution as the solution for system suitability test.Pipet 1 mL of the solution, and add the mobile phase tomake exactly 10 mL. Conˆrm that the peak area of digoxinobtained from 10 mL of this solution is equivalent to 7 to13z of that from the solution for system suitability test.
System performance: Dissolve 25 mg of digoxin in 50 mLof warm ethanol (95), cool, and add ethanol (95) to makeexactly 100 mL. Pipet 10 mL of this solution, add exactly5 mL of a solution of propyl parahydroxybenzoate inethanol (95) (1 in 4000), 10 mL of water and dilute ethanol tomake 50 mL. When the procedure is run with 10 mL of thissolution under the above operating conditions, digoxin andpropyl parahydroxybenzoate are eluted in this order with theresolution between these peaks being not less than 5.
System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of digoxin is not more than 1.0z.
Change the Assay to read:
Assay Weigh accurately about 25 mg each of Digoxin andDigoxin Reference Standard, previously dried, dissolve in50 mL of warm ethanol (95), cool, and add ethanol (95) tomake exactly 100 mL. Pipet 10 mL of these solutions, addexactly 5 mL of the internal standard solution, 10 mL ofwater and dilute ethanol to make 50 mL, and use thesesolutions as the sample solution and the standard solution.Perform the test with 10 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area of digoxinto that of the internal standard.
Amount (mg) of C41H64O14＝WS×QT
QS
WS: Amount (mg) of Digoxin Reference Standard
Internal standard solution—A solution of propyl para-hydroxybenzoate in ethanol (95) (1 in 4000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 220 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about
17031703Supplement II, JPXIV O‹cial Monographs for Part I
309C.Mobile phase: A mixture of water and acetonitrile (7:3).Flow rate: Adjust the ‰ow rate so that the retention time
of digoxin is about 10 minutes.System suitability—
System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, digoxin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 5.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of digoxin to that of the internal standard isnot more than 1.0z.
Change to read:
Digoxin Injectionジゴキシン注射液
Digoxin Injection is an aqueous solution forinjection.
It contains not less than 90.0z and not more than105.0z of the labeled amount of digoxin (C41H64O14:780.94).
Method of preparation Prepare as directed under Injec-tions, with a solution of Digoxin in 5 to 50 volz ethanol.
Description Digoxin Injection is a clear, colorless liquid.
Identiˆcation Dilute Digoxin Injection, if necessary, withmethanol so that each mL contains about 0.25 mg of Digox-in according to the labeled amount, and use this solution asthe sample solution. In case where ingredients are suspectedto aŠect the test, remove them by means of a solid-phase ex-traction. Separately, dissolve 0.5 mg of Digoxin ReferenceStandard in 2 mL of methanol, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 10 mLeach of the sample solution and the standard solution on aplate of octadecylsilanized silica gel for thin-layer chro-matography. Develop the plate with a mixture of methanoland water (7:3) to a distance of about 10 cm, and air-dry theplate. Spray evenly a mixture of a solution of trichloroaceticacid in ethanol (99.5) (1 in 4) and a freshly prepared solutionof sodium toluenesulfonchloramide trihydrate (3 in 100)(4:1) on the plate, heat at 1109C for 10 minutes, andexamine under ultraviolet light (main wavelength: 366 nm):the Rf values of the principal spots with the sample solutionand the standard solution are not diŠerent each other.
Bacterial endotoxins Less than 200 EU/mg.
Actual volume It meets the requirements of Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirements of the Foreign Insoluble
Matter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirement of the Insoluble Particu-late Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay To an exact volume of Digoxin Injection, equivalentto about 2.5 mg of digoxin (C41H64O14), add exactly 5 mL ofthe internal standard solution and dilute ethanol to make50 mL, and use this solution as the sample solution.Separately, weigh accurately about 25 mg of Digoxin Refer-ence Standard, previously dried under reduced pressure at1059C for 1 hour, dissolve in 50 mL of warm ethanol (95),cool, and add ethanol (95) to make exactly 100 mL. Pipet 10mL of this solution, add exactly 5 mL of the internal stan-dard solution, 10 mL of water and dilute ethanol to make 50mL, and use this solution as the standard solution. Performthe test with 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of digoxin to that of theinternal standard.
Amount (mg) of digoxin (C41H64O14)＝WS×QT
QS×
110
WS: Amount (mg) of Digoxin Reference Standard
Internal standard solution—A solution of propyl para-hydroxybenzoate in ethanol (95) (1 in 4000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 220 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about309C.
Mobile phase: A mixture of water and acetonitrile (7:3).Flow rate: Adjust the ‰ow rate so that the retention time
of digoxin is about 10 minutes.System suitability—
System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, digoxin and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 5.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the aboveoperating conditions, the relative standard deviation of theratio of the peak area of digoxin to that of the internalstandard is not more than 1.0z.
Containers and storage Containers—Hermetic containers,and colored containers may be used.
Storage—Light-resistant.
17041704 Supplement II, JPXIVO‹cial Monographs for Part I
Change to read:
Digoxin Tabletsジゴキシン錠
Digoxin Tablets contain not less than 90.0z and notmore than 105.0z of the labeled amount of digoxin(C41H64O14: 780.94).
Method of preparation Prepare as directed under Tablets,with Digoxin.
Identiˆcation To an amount of pulverized DigoxinTablets, equivalent to 0.5 mg of Digoxin according to the la-beled amount, add 2 mL of methanol, shake for 10 minutes,ˆlter, and use the ˆltrate as the sample solution. Separately,dissolve 0.5 mg of Digoxin Reference Standard in 2 mL ofmethanol, and use this as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of octadecylsilanizedsilica gel for thin-layer chromatography. Develop the platewith a mixture of methanol and water (7:3) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly a mixture ofa solution of trichloroacetic acid in ethanol (99.5) (1 in 4)and a freshly prepared solution of sodium toluenesulfon-chloramide trihydrate (3 in 100) (4:1) on the plate, heat at1109C for 10 minutes, and examine under ultraviolet light(main wavelength: 366 nm): the Rf values of the principalspots with the sample solution and the standard solution arenot diŠerent each other.
Content uniformity Perform the test according to the fol-lowing method: it meets the requirements of the ContentUniformity Test.
To 1 tablet of Digoxin Tablets add 0.5 mL of water todisintegrate, then add exactly 0.5 mL of the internal stan-dard solution, and add V mL of dilute ethanol so that eachmL contains about 21 mg of digoxin (C41H64O14). Exposurethis solution to ultrasonic waves for 20 minutes, shake for 5minutes, ˆlter, and use the ˆltrate as the sample solution.Separately, weigh accurately about 25 mg of Digoxin Refer-ence Standard, previously dried under reduced pressure at1059C for 1 hour, dissolve in 50 mL of warm ethanol (95),cool, and add ethanol (95) to make exactly 100 mL. Pipet10 mL of this solution, and add ethanol (95) to make exactly20 mL. Pipet 1 mL of this solution, add exactly 0.5 mL ofthe internal standard solution, then add 1.5 mL of water and(V – 2) mL of dilute ethanol, and use this as the standardsolution. Proceed with the sample solution and the standardsolution as directed in the Assay.
Amount (mg) of digoxin (C41H64O14)＝WS×QT
QS×
1200
WS: Amount (mg) of Digoxin Reference Standard
Internal standard solution—A solution of propyl para-hydroxybenzoate in ethanol (95) (1 in 40,000/V).
Dissolution Perform the test with 1 tablet of DigoxinTablets, using 500 mL of diluted hydrochloric acid (3 in 500)at 100 revolutions per minute according to the Method 1 un-der the Dissolution Test as the dissolution medium.Withdraw 30 mL or more of the dissolved solution 60minutes after starting the test, and ˆlter through a mem-brane ˆlter (less than 0.8 mm in pore size). Discard the ˆrst10 mL of the ˆltrate, and use the subsequent ˆltrate as thesample solution. Separately, weigh accurately about 25 mgof Digoxin Reference Standard, previously dried in vacuumat 1059C for 1 hour, dissolve in a small portion of ethanol(95), and add a mixture of ethanol (95) and water (4:1) tomake exactly 500 mL. Pipet 5 mL of this solution, add thedissolution medium to make exactly 500 mL, and use this so-lution as the standard solution. Pipet 2 mL each of the sam-ple solution, the standard solution and the dissolution medi-um, and transfer to brown glass-stoppered test tubes. Addexactly 10 mL of 0.012 g/dL L-ascorbic acid-hydrochloricacid TS to these tubes, and shake. Immediately add exactly 1mL of dilute hydrogen peroxide TS, shake well, and allow tostand at a constant temperature between 259C and 309C for45 minutes. Determine the ‰uorescence intensities, FT, FS,and FB, of these solutions at 360 nm of the excitationwavelength and at 485 nm of the ‰uorescence wavelength asdirected under the Fluorometry, respectively: the dissolutionrate in 60 minutes is not less than 65z. No retest require-ment is applied to Digoxin Tablets.
Dissolution rate (z) with respect to the labeled amount ofdigoxin (C41H64O14)
＝WS×FT－FB
FS－FB×
1C
WS: Amount (mg) of Digoxin Reference StandardC: The labeled amount (mg) of digoxin (C41H64O14) in 1
tablet
Assay Weigh accurately the mass of not less than 20Digoxin Tablets, and powder. Weigh accurately a portion ofthe powder, equivalent to about 2.5 mg of digoxin (C41H64O
14), add 30 mL of dilute ethanol, exposure to ultrasonicwaves for 20 minutes, and shake for 5 minutes. After cool-ing, add exactly 5 mL of the internal standard solution anddilute ethanol to make 50 mL, centrifuge, and use the super-natant liquid as the sample solution. Separately, weigh ac-curately about 25 mg of Digoxin Reference Standard, previ-ously dried under reduced pressure at 1059C for 1 hour , dis-solve in 50 mL of warm ethanol (95), cool, and add ethanol(95) to make exactly 100 mL. Pipet 10 mL of this solution,add exactly 5 mL of the internal standard solution, 10 mL ofwater and dilute ethanol to make 50 mL, and use this solu-tion as the standard solution. Perform the test with 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of digoxin to that of the internal standard.
Amount (mg) of digoxin (C41H64O14)＝WS×QT
QS×
110
17051705Supplement II, JPXIV O‹cial Monographs for Part I
WS: Amount (mg) of Digoxin Reference Standard
Internal standard solution—A solution of propyl para-hydroxybenzoate in ethanol (95) (1 in 4000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 220 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about309C.
Mobile phase: A mixture of water and acetonitrile (7:3).Flow rate: Adjust the ‰ow rate so that the retention time
of digoxin is about 10 minutes.System suitability—
System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, digoxin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 5.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of digoxin to that of the internal standard isnot more than 1.0z.
Containers and storage Containers—Tight containers.Storage—Light-resistant.
Dimorpholamineジモルホラミン
Change the origin/limits of the content to read:
Dimorpholamine, when dried, contains not less than98.0z and not more than 101.0z of C20H38N4O4.
Change the Description to read:
Description Dimorpholamine is a white to light yellow,crystalline powder, masses or syrupy liquid.
It is very soluble in ethanol (99.5) and in acetic anhydride,and soluble in water.
The pH of a solution prepared by dissolving 1.0 g ofDimorpholamine in 10 mL of water is between 6.0 and 7.0.
It is hygroscopic.
Change the Identiˆcation to read:
Identiˆcation (1) Determine the absorption spectrum ofa solution of Dimorpholamine (1 in 50,000) as directedunder the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewavelengths.
(2) Determine the infrared absorption spectrum of
Dimorpholamine, previously dried, as directed in the potas-sium bromide disk method under the Infrared Spectrophoto-metry, and compare the spectrum with the ReferenceSpectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
Add the following next to Purity (4):
(5) Related substances—Dissolve 0.20 g of Dimorphola-mine in 10 mL of ethanol (99.5), and use this solution as thesample solution. Pipet 1 mL of the sample solution, addethanol (99.5) to make exactly 100 mL, and use this solutionas the standard solution. Perform the test with these solu-tions as directed under the Thin-layer Chromatography.Spot 10 mL each of the sample solution and the standardsolution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethanol(99.5) and water (4:1) to a distance of about 10 cm, andair-dry the plate. Allow the plate to stand in iodine vapor for10 minutes: the spot other than the principal spot from thesample solution is not more intense than the spot from thestandard solution.
Change the Assay to read:
Assay Weigh accurately about 0.6 g of Dimorpholamine,previously dried, dissolve in 50 mL of acetic anhydride, andtitrate with 0.1 mol/L perchloric acid VS (potentiometrictitration). Perform a blank determination, and make anynecessary correction.
Each mL of 0.1 mol/L perchloric acid VS＝39.85 mg of C20H38N4O4
Dimorpholamine Injectionジモルホラミン注射液
Change the origin/limits of content to read:
Dimorpholamine Injection is an aqueous solutionfor injection.
It contains not less than 95.0z and not more than105.0z of the labeled amount of dimorpholamine(C20H38N4O4: 398.54).
Change the Description to read:
Description Dimorpholamine Injection is a clear, colorlessliquid.
pH: 3.0 – 5.5
Change the Identiˆcation (2) to read:
Identiˆcation(2) To a volume of Dimorpholamine Injection, equiva-
lent to 50 mg of Dimorpholamine according to the labeledamount, add 1 mL of dilute hydrochloric acid, andevaporate on a water bath to dryness. Dissolve the residue in
17061706 Supplement II, JPXIVO‹cial Monographs for Part I
2 mL of hydrochloric acid, boil for 10 minutes under a re‰uxcondenser, and evaporate to dryness on a water bath.Dissolve the residue with 1 mL of water, neurtralize withsodium hydroxide TS, and add 0.2 mL of a solution ofacetaldehyde (1 in 20), 0.1 mL of sodium pentacyanonitrosylferrate (III) TS and 0.5 mL of sodium carbonate TS: a bluecolor develops.
Add the following next to Identiˆcation:
Bacterial endotoxins Less than 5.0 EU/mg. Perform thetest with the sample diluted to 0.15 w/vz with water forbacterial endotoxins test.
Actual volume It meets the requirements of Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Change the Assay to read:
Assay Measure exactly a volume of DimorpholamineInjection, equivalent to about 30 mg of dimorpholamine(C20H38N4O4), and add water to make exactly 200 mL. Pipet1 mL of this solution, shake with exactly 4 mL of the inernalstandard solution for 5 minutes, and use this solution as thesample solution. Separately, weigh accurately about 0.15 gof dimorpholamine for assay, previously dried in a desicca-tor (in vacuum, phosphorus (V) oxide) for 8 hours, anddissolve in water to make exactly 1000 mL. Pipet 1 mL ofthis solutionn, shake with exactly 4 mL of the inernalstandard solution for 5 minutes, and use this solution as thestandard solution. Perform the test with 10 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and calculate the ratios, QT and QS, of the peakarea of dimorpholamine to that of the inernal standard.
Amount (mg) of dimorpholamine (C20H38N4O4)
＝WS×QT
QS×
15
WS: Amount (mg) of dimorpholamine for assay
Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 25,000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 216 nm).
Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about409C.
Mobile phase: A mixture of water and acetonitrile (1:1).Flow rate: Adjust the ‰ow rate so that the retention time
of dimorpholamine is about 4 minutes.System suitability—
System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, dimorpholamine and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of dimorpholamine to that of the internalstandard is not more than 1.0z.
Eperisone Hydrochloride contains not less than98.5z and not more than 101.0z of C17H25NO.HCl,calculated on the anhydrous basis.
Description Eperisone Hydrochloride occurs as a whitecrystalline powder.
It is freely soluble in water, in methanol and in acetic acid(100), and soluble in ethanol (99.5).
Melting point: about 1679C (with decomposition).A solution of Eperisone Hydrochloride in methanol (1 in
100) shows no optical rotation.
Identiˆcation (1) Determine the absorption spectrum ofa solution of Eperisone Hydrochloride in methanol (1 in100,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities ofabsorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofEperisone Hydrochloride as directed in the potassium chlo-ride disk method under the Infrared Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.
(3) A solution of Eperisone Hydrochloride (1 in 50)responds to the Qualitative Tests for chloride.
Purity (1) Heavy metals—Proceed with 1.0 g of Eperi-
17071707Supplement II, JPXIV O‹cial Monographs for Part I
sone Hydrochloride according to Method 1, and perform thetest. Prepare the control solution with 2.0 mL of StandardLead Solution (not more than 20 ppm).
(2) Piperidine hydrochloride—Dissolve 1.0 g of Eperi-sone Hydrochloride in 20 mL of water, add 2.0 mL ofdiluted hydrochloric acid (1 in 2), 2.0 mL of a solution ofcopper (II) sulfate pentahydrate (1 in 20) and 1.5 mL of am-monia solution (28), and use this solution as the sample solu-tion. Separately, to 2.0 mL of a solution of piperidinehydrochloride (1 in 1000) add 18 mL of water, 2.0 mL ofdiluted hydrochloric acid (1 in 2), 2.0 mL of a solution ofcopper (II) sulfate pentahydrate (1 in 20) and 1.5 mL ofammonia solution (28), and use this solution as the standardsolution. To each of the sample solution and the standardsolution add 10 mL of a mixture of isopropylether andcarbon disulˆde (3:1), shake for 30 seconds, allow them tostand for 2 minutes, and compare the color of the upperlayer: the color obtained from the sample solution is notmore darker than that from the standard solution.
(3) Related substances—Dissolve 0.1 g of EperisoneHydrochloride in 100 mL of the mobile phase, and use thissolution as the sample solution. Pipet 1 mL of the samplesolution, add the mobile phase to make exactly 100 mL, anduse this solution as the standard solution. Perform the testwith exactly 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the totalarea of the peaks other than the peak of eperisone is notmore than 1/5 of the peak area of eperisone from the stan-dard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).Column: A stainless steel column 4.6 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about309C.
Mobile phase: A mixture of methanol, 0.0375 mol/L sodi-um 1-decanesulfonate TS and perchloric acid (600:400:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof eperisone is about 17 minutes.
Time span of measurement: About 2 times as long as theretention time of eperisone.System suitability—
Test for required detectability: Pipet 1 mL of the standardsolution, and add the mobile phase to make exactly 10 mL.Conˆrm that the peak area of eperisone obtained from 10mL of this solution is equivalent to 7 to 13z of that obtainedfrom 10 mL of the standard solution.
System performance: When the procedure is run with 10mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of eperisone are not less than 4000 stepsand not more than 2.0, respectively.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-
ing conditions, the relative standard deviation of the peakarea of eperisone is not more than 3.0z.
Water Not more than 0.2z (0.1 g, coulometric titration).
Residue on ignition Not more than 0.2z (1 g).
Assay Weigh accurately about 0.6 g of EperisoneHydrochloride, dissolve in 20 mL of acetic acid (100), add80 mL of acetic anhydride, and titrate with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blankdetermination in the same manner, and make any necessarycorrection.
Each mL of 0.1 mol/L perchloric acid VS＝29.58 mg of C17H25NO.HCl
Containers and storage Containers—Well-closed contain-ers.
Ethionamideエチオナミド
Change to read except the structural formulaand chemical name:
Ethionamide, when dried, contains not less than98.5z and not more than 101.0z of C8H10N2S.
Description Ethionamide occurs as yellow crystals or crys-talline powder, having a characteristic odor.
It is soluble in methanol and in acetic acid (100), sparinglysoluble in ethanol (99.5) and in acetone, and practically in-soluble in water.
Identiˆcation (1) Determine the absorption spectrum ofa solution of Ethionamide in methanol (3 in 160,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.
(2) Determine the infrared absorption spectrum ofEthionamide as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.
Melting point 161 – 1659C
Purity(1) Acid—Dissolve 3.0 g of Ethionamide in 30 mL of
methanol by warming, add 90 mL of water, allow to stand inice water for 1 hour, and ˆlter. To 80 mL of the ˆltrate add0.8 mL of cresol red TS and 0.20 mL of 0.1 mol/L sodiumhydroxide VS: a red color develops.
(2) Heavy metals—Proceed with 1.0 g of Ethionamideaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).
17081708 Supplement II, JPXIVO‹cial Monographs for Part I
(3) Arsenic—Prepare the test solution with 1.0 g ofEthionamide according to Method 3. Add 10 mL of a solu-tion of magnesium nitrate hexahydrate in ethanol (95) (1 in50), then add 1.5 mL of hydrogen peroxide (30), and ˆre toburn (not more than 2 ppm).
(4) Related substances—Conduct this procedure withoutexposure to light, using light-resistant vessels. Dissolve 0.20g of Ethionamide in 10 mL of acetone, and use this solutionas the sample solution. Pipet 0.5 mL of the sample solution,add acetone to make exactly 100 mL, and use this solution asthe standard solution (1). Separately, pipet exactly 0.2 mLof the sample solution, add acetone to make exactly 100 mL,and use this solution as the standard solution (2). Performthe test with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solutions (1) and (2) on a plate of silica gelwith ‰uorescent indicator for thin-layer chromatography,develop with a mixture of ethyl acetate, hexane andmethanol (6:2:1) to a distance of about 15 cm, and air-drythe plate. Examine under ultraviolet light (main wavelength:254 nm): the spot other than the principal spot obtained withthe sample solution is not more intense than the spot withthe standard solution (1), and number of the spot other thanthe principal spot obtained with the sample solution which ismore intense than the spot with the standard solution (2) isnot more than one.
Loss on drying Not more than 0.5z (1 g, 1059C, 3 hours).
Residue on ignition Not more than 0.10z (1 g).
Assay Weigh accurately about 0.3 g of Ethionamide,previously dried, dissolve in 50 mL of acetic acid (100), andtitrate with 0.1 mol/L perchloric acid VS until the color ofthe solution changes from orange-red to dark orange-brown(indicator: 2 mL of p-naphtholbenzein TS). Perform a blankdetermination in the same manner, and make any necessarycorrection.
Each mL of 0.1 mol/L perchloric acid VS＝16.62 mg of C8H10N2S
Containers and storage Containers—Well-closed contain-ers.
Etilefrine Hydrochloride Tablets塩酸エチレフリン錠
Change the origin/limits of content to read:
Etilefrine Hydrochloride Tablets contain not lessthan 93.0z and not more than 107.0z of the labeledamount of etilefrine hydrochloride (C10H15NO2.HCl:217.69).
Change the Identiˆcation to read:
Identiˆcation To a quantity of powdered EtilefrineHydrochloride Tablets, equivalent to 5 mg of EtilefrineHydrochloride according to the labeled amount, add 60 mLof diluted hydrochloric acid (1 in 1000), shake well, add40 mL of diluted hydrochloric acid (1 in 1000), and ˆlter.Determine the absorption spectrum of the ˆltrate as directedunder the Ultraviolet-visible Spectrophotometry, usingdiluted hydrochloric acid (1 in 1000) as the blank: it exhibitsa maximum between 271 nm and 275 nm.
Add the following next to Identiˆcation:
Content uniformity Perform the test according to the fol-lowing method: it meets the requirements of the ContentUniformity Test.
To 1 tablet of Etilefrine Hydrochloride Tablets add 60 mLof diluted hydrochloric acid (1 in 1000), and proceed asdirected in the Assay.
Amount (mg) of etilefrine hydrochloride (C10H15NO2.HCl)
＝WS×AT
AS×
110
WS: Amount (mg) of etilefrine hydrochloride for assay
Change the Assay to read:
Assay Weigh accurately the mass of not less than 20Etilefrine Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 5 mgof etilefrine hydrochloride (C10H15NO2.HCl), add 60 mL ofdiluted hydrochloric acid (1 in 1000), shake for 10 minutes,add diluted hydrochloric acid (1 in 1000) to make exactly 100mL, and ˆlter. Discard the ˆrst 20 mL of the ˆltrate, and usethe subsequent ˆltrate as the sample solution. Separately,weigh accurately about 50 mg of etilefrine hydrochloride forassay, previously dried at 1059C for 4 hours, and dissolve indiluted hydrochloric acid (1 in 1000) to make exactly 100mL. Pipet 10 mL of this solution, add diluted hydrochloricacid (1 in 1000) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly20 mL each of the sample solution and the standard solutionas direct under the Liquid Chromatography according to thefollowing conditions, and determine the peak areas, AT andAS, of etilefrine.
Amount (mg) of etilefrine hydrochloride (C10H15NO2.HCl)
＝WS×AT
AS×
110
WS: Amount (mg) of etilefrine hydrochloride for assay
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 220 nm).Column: A stainless steel column 4.6 mm in inside di-
ameter and 25 cm in length, packed with octylsilanized silicagel for liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about409C.
17091709Supplement II, JPXIV O‹cial Monographs for Part I
Mobile phase: Dissolve 5 g of sodium lauryl sulfate in940 mL of water and 500 mL of acetonitrile, and adjust thepH to 2.3 with phosphoric acid.
Flow rate: Adjust the ‰ow rate so that the retention timeof etilefrine is about 6 minutes.System suitability—
System performance: Dissolve 4 mg of bamethan sulfateand 4 mg of etilefrine hydrochloride in the mobile phase tomake 50 mL. When the procedure is run with 20 mL of thissolution under the above operating conditions, etilefrine andbamethan are eluted in this order with the resolutionbetween these peaks being not less than 5.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of etilefrine is not more than 1.0z.
Etoposide contains not less than 98.0z and notmore than 102.0z of C29H32O13, calculated on theanhydrous basis.
Description Etoposide occurs as white crystals or crystal-line powder.
It is sparingly soluble in methanol, slightly soluble inethanol (99.5), and very slightly soluble in water.
Melting point: about 2609C (with decomposition).
Identiˆcation (1) Determine the absorption spectrum ofa solution of Etoposide in methanol (1 in 10,000) as directedunder the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of a solution of Etoposide Reference Standardprepared in the same manner as the sample solution: bothspectra exhibit similar intensities of absorption at the samewavelengths.
(2) Determine the infrared absorption spectrum of
Etoposide as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofEtoposide Reference Standard: both spectra exhibit similarintensities of absorption at the same wave numbers.
Purity (1) Heavy metals—Proceed with 2.0 g of Etopo-side according to Method 2, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 10 ppm).
(2) Related substances—Dissolve 50 mg of Etoposide in10 mL of methanol, add the mobile phase to make 50 mL,and use this solution as the sample solution. Pipet 2 mL ofthe sample solution, add the mobile phase to make exactly200 mL, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solutionand the standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine each peak area by the automatic integrationmethod: the area of the peak other than etoposide is notlarger than 1/5 times the peak area of etoposide with thestandard solution, and the total area of the peaks other thanthe peak of etoposide with the sample solution is not largerthan 1/2 times the peak area of etoposide with the standardsolution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 3 times as long as theretention time of etoposide after the solvent peak.System suitability—
Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of etoposideobtained with 50 mL of this solution is equivalent to 7 to13z of that with 50 mL of the standard solution.
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of etoposide is not more than 2.0z.
Water Not more than 4.0z (0.5 g, volumetric titration,direct titration).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately about 25 mg each of Etoposideand Etoposide Reference Standard, separately determinedthe water content, and dissolve separately in methanol tomake exactly 25 mL. Pipet 10 mL each of these solutions,add exactly 5 mL of the internal standard solution and themobile phase to make 50 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith 50 mL each of the sample solution and the standard
17101710 Supplement II, JPXIVO‹cial Monographs for Part I
solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of etoposide to that ofthe internal standard.
Amount (mg) of C29H32O13＝WS×QT
QS
WS: Amount (mg) of Etoposide Reference Standard,calculated on the anhydrous basis
Internal standard solution—A solution of 2,6-dichloro-phenol in methanol (3 in 2500).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 290 nm).
Column: A stainless steel column 3.9 mm in insidediameter and 30 cm in length, packed with phenylsilanizedsilica gel for liquid chromatography (10 mm in particlediameter).
Column temperature: A constant temperature of about359C.
Mobile phase: Dissolve 6.44 g of sodium sulfate decahy-drate in diluted acetic acid (100) (1 in 100) to make 1000 mL,and add 250 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof etoposide is about 20 minutes.System suitability—
System performance: Dissolve 10 mg of Etoposide in2 mL of methanol, add 8 mL of the mobile phase, and mixwell. Add 0.1 mL of diluted acetic acid (100) (1 in 25) and0.1 mL of phenolphthalein TS, and add sodium hydroxideTS until the color of the solution changes to faintly red. Af-ter allowing to stand for 15 minutes, add 0.1 mL of dilutedacetic acid (100) (1 in 25). When the procedure is run with 10mL of this solution under the above operating conditions, theresolution between the peak of etoposide and the peak hav-ing the relative retention time of about 1.3 with respect toetoposide is not less than 3.
System repeatability: When the test is repeated 6 timeswith 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of etoposide to that of the internal standard isnot more than 1.0z.
Containers and storage Containers—Tight containers.Storage—Light-resistant.
Purity(5) Related substances—Dissolve 0.10 g of Flavin Ade-
nine Dinucleotide Sodium in 200 mL of the mobile phase,and use this solution as the sample solution. Perform the test
with 20 mL of the sample solution as directed under theLiquid Chromatography according to the following condi-tions. Determine the peak area, A, of ‰avin adeninedinucleotide and the total area, S, of peaks other than thepeak of ‰avin adenine dinucleotide by the automatic integra-tion method: S/(A＋S) is not more than 0.10.Operating conditions—
Column, column temperature, mobile phase, ‰ow rate,and time span of measurement: Proceed as directed in theoperating conditions in the Procedure (ii) under Assay (1).
Detector: An ultraviolet absorption photometer(wavelength: 260 nm).System suitability—
Test for required detection: To exactly 2 mL of the samplesolution add the mobile phase to make exactly 20 mL, anduse this solution as the solution for system suitability test.Conˆrm that the peak area of ‰avin adenine dinucleotideobtained from 20 mL of this solution is equivalent to 8 to12z of that of ‰avin adenine dinucleotide obtained from20 mL of the sample solution.
System performance: Proceed as directed in the systemsuitability in the Procedure (ii) under Assay (1).
System repeatability: When the test is repeated 6 timeswith 20 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of ‰avin adenine dinucleotide is notmore than 1.0z.
Change the Assay (1) to read:
Assay (1) Procedure (i) Total ‰avin content—Conductthis procedure without exposure to daylight, using light-resistant vessels. Weigh accurately about 0.1 g of FlavinAdenine Dinucleotide Sodium, and dissolve in water tomake exactly 200 mL. Pipet 5 mL of this solution, add 5 mLof zinc chloride TS, and heat in a water bath for 30 minutes.After cooling, add water to make exactly 100 mL, and usethis solution as the sample solution. Separately, weighaccurately about 50 mg of Ribo‰avin Reference Standard,previously dried at 1059C for 2 hours, dissolve in 200 mL ofdiluted acetic acid (100) (1 in 100) by warming, cool, addwater to make exactly 500 mL. Pipet 10 mL of this solution,add water to make exactly 100 mL, and use this solution asthe standard solution. Determine the absorbances, AT and AS, of the sample solution and the standard solution at 450 nmas directed under the Ultraviolet-visible Spectrophotometry,using water as the blank.
Total amout (mg) of ‰avin
＝WS×AT
AS×
45
WS: Amount (mg) of Ribo‰avin Reference Standard
(ii) Peak area ratio of ‰avin adenine dinucleotide—Dis-solve 0.1 g of Flavin Adenine Dinucleotide Sodium in 200mL of water, and use this solution as the sample solution.Perform the test with 5 mL of this solution as directed underthe Liquid Chromatography according to the followingconditions. Determine the peak area, A of ‰avin adenine
17111711Supplement II, JPXIV O‹cial Monographs for Part I
dinucleotide, and the total area, S, of the peaks other than‰avin adenine dinucleotide by the automatic integrationmethod.
Peak area ratio of ‰avin adenine dinucleotide
＝1.08×A
1.08×A＋S
Operating conditions—Detector: A visible spectrophotometer (wavelength: 450
nm).Column: A stainless steel column 4 mm in inside diameter
and 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about359C.
Mobile phase: A mixture of a solution of potassium di-hydrogen phosphate (1 in 500) and methanol (4:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof ‰avin adenine dinucleotide is about 10 minutes.
Time span of measurement: About 4.5 times as long as theretention time of ‰avin adenine dinucleotide.System suitability—
Test for required detection: To exactly 2 mL of the samplesolution add water to make exactly 20 mL, and use this solu-tion as the solution for system suitability test. Pipet 2 mL ofthe solution, and add water to make exactly 20 mL. Conˆrmthat the peak area of ‰avin adenine dinucleotide obtainedfrom 5 mL of this solution is equivalent to 8 to 12z of thatof ‰avin adenine dinucleotide obtained from 5 mL of thesolution for system suitability test.
System performance: Dissolve 20 mg each of FlavinAdenine Dinucleotide Sodium and ribo‰avin sodium phos-phate in 100 mL of water. When the procedure is run with5 mL of this solution under the above operating conditions,‰avin adenine dinucleotide and ribo‰avin phosphate areeluted in this order with the resolution between these peaksbeing not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 5 mL of the solution for system suitability test under theabove operating conditions, the relative standard deviationof the peak area of ‰avin adenine dinucleotide is not morethan 1.0z.
(2) Calculation
Amount (mg) of C27H31N9Na2O15P2
＝fT×fR×2.2040
fT: Total amount (mg) of ‰avin in Flavin AdenineDinucleotide Sodium obtained from the procedure (i).
fR: Peak area ratio of ‰avin adenine dinucleotide in FlavinAdenine Dinucleotide Sodium obtained from theprocedure (ii).
Add the following:
Flomoxef Sodium for Injection注射用フロモキセフナトリウム
Flomoxef Sodium for Injection is a preparation forinjection which is dissolved before use.
It contains not less than 90.0z and not morethan 110.0z of the labeled amount of ‰omoxef(C15H18F2N6O7S2: 496.47).
Method of preparation Prepare as directed under Injec-tions, with Flomoxef Sodium.
Description Flomoxef Sodium for Injection occurs aswhite to light yellowish white, friable masses or powder.
Identiˆcation Proceed as directed in the Identiˆcation (3)under Flomoxef Sodium.
pH The pH of a solution obtained by dissolving an amountof Flomoxef Sodium for Injection, equivalent to 0.5 g(potency) of Flomoxef Sodium according to the labeledamount, in 5 mL of water is between 4.0 and 5.5.
Purity (1) Clarity and color of solution—Dissolve anamount of Flomoxef Sodium for Injection, equivalent to 1.0g (potency) of Flomoxef Sodium according to the labeledamount, in 10 mL of water: the solution is clear and color-less or pale yellow.
(2) 1-(2-Hydroxyethyl)-1H-tetrazol-5-thiol—Use thesample solution obtained in the Assay as the sample solu-tion. Weigh accurately about 20 mg of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol, and dissolve in water to make exactly100 mL. Pipet 5 mL of this solution, add exactly 25 mL ofthe internal standard solution and water to make 50 mL, anduse this solution as the standard solution. Perform the testwith 5 mL each of the sample solution and the standard solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the ratios, QT
and QS, of the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol to that of the internal standard. Calculate the amountof 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol per 1 g (potency)of Flomoxef Sodium for Injection by the following formula:not more than 10 mg.
Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol(C3H6N4OS)
＝WS×QT
QS×
110
WS: Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol
Operating conditions—Proceed as directed in the Assay.
System suitability—Test for required detectability: Pipet 1 mL of the standard
solution, and add water to make exactly 20 mL. Conˆrmthat the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiolobtained from 5 mL of this solution is equivalent to 3.5 –6.5z of that obtained from 5 mL of the standard solution.
17121712 Supplement II, JPXIVO‹cial Monographs for Part I
System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol and the inter-nal standard are eluted in this order with the resolutionbetween these peaks being not less than 20.
System repeatability: When the test is repeated 3 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol tothat of the internal standard is not more than 1.0z.
Water Not more than 1.5z (0.5 g, volumetric titration,back titration).
Bacterial endotoxins Less than 0.025 EU/mg (potency).
Mass variation It meets the requirements of the MassVariation Test.
Foreign insoluble matter Perform the test according toMethod 2: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay Weigh accurately the mass of the contents of notless than 10 Flomoxef Sodium for Injection, and calculatethe average mass of the content. Spread out thinly about 1 gof the content in a petri dish, allow the dish to stand in adesiccator containing a saturated solution of magnesiumbromide without light exposure to equilibrate the sample toconstant water content. Determine the water content,separately, with about 0.1 g of the sample according to themethod described in Water. Weigh accurately an amount ofthe sample, equivalent to about 50 mg (potency) of Flomox-ef Sodium according to the labeled amount, add exactly 50mL of the internal standard solution to dissolve, add waterto make 100 mL, and use this solution as the sample solu-tion. Separately weigh accurately about 50 mg (potency) ofFlomoxef Triethylammonium Reference Standard, addexactly 50 mL of the internal standard solution to dissolve,add water to make 100 mL, and use this solution as thestandard solution. Proceed as directed in the Assay underFlomoxef Sodium.
Amount [mg (potency)] of ‰omoxef (C15H18F2N6O7S2)
＝WS×QT
QS×1000
WS: Amount [mg (potency)] of Flomoxef Triethylammo-nium Reference Standard
Internal standard solution—A solution of m-cresol (3 in1000).
Containers and storage Containers—Hermetic containers.Polyethylene or polypropylene containers for aqueous
injection may be used.
Add the following:
Flopropione Capsulesフロプロピオンカプセル
Flopropione Capsules contain not less than 93.0zand not more than 107.0z of the labeled amount of‰opropione (C9H10O4: 182.17).
Method of preparation Prepare as directed under theCapsules, with Flopropione.
Identiˆcation (1) Powder the contents of FlopropioneCapsules. To a portion of the powder, equivalent to 60 mgof Flopropione according to the labeled amount, add 40 mLof water, shake well, and ˆlter. To 5 mL of the ˆltrate add1 mL of iron (III) nitrate TS: a red-purple color appears.
(2) Powder the contents of Flopropione Capsules. To aportion of the powder, equivalent to 90 mg of Flopropioneaccording to the labeled amount, add 100 mL of ethanol(99.5), shake well, and ˆlter. To 5 mL of the ˆltrate addethanol (99.5) to make 50 mL. To 5 mL of this solution addethanol (99.5) to make 100 mL, and use this solution as thesample solution. Determine the absorption spectrum of thesample solution as directed under the Ultraviolet-visibleSpectrophotometry: it exhibits a maximum between 283 nmand 287 nm.
Content uniformity Perform the test as directed in theAssay, and determine the content: it meets the requirementsof the Content Uniformity Test.
Dissolution Being speciˆed separately.
Assay To 1 capsule of Flopropione Capsules add 43 mL ofa mixture of water and phosphoric acid (86:1), and disinte-grate the capsule at 509C in a water bath. After cooling, addacetonitrile to make exactly 100 mL, stir for 10 minutes, andcentrifuge a part of this solution for 5 minutes at 3000 rpm.Use the supernatant liquid as the sample solution. Separate-ly, weigh accurately about 40 mg of ‰opropione for assay,separately determine the water content in the same manneras Flopropione, add 70 mL of the mobile phase, and dis-solve by exposure for 10 minutes to ultrasonic vibration.Add the mobile phase to make exactly 100 mL, and use thissolution as the standard solution. Perform the test withexactly 5 mL each of the sample solution and the standardsolution as directed under the Liquid chromatography ac-cording to the following conditions, and determine the peakareas, AT and AS, of ‰opropione. Repeat the procedureabove with totally not less than 10 capsules, and calculatethe mean.
Amount (mg) of ‰opropione (C9H10O4)＝WS×AT
AS
WS: Amount (mg) of ‰opropione for assay, calculated on
17131713Supplement II, JPXIV O‹cial Monographs for Part I
the anhydrous basis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 267 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about359C.
Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (114:86:1)
Flow rate: Adjust the ‰ow rate so that the retention timeof ‰opropione is about 3 minutes.System suitability—
System performance: Dissolve 50 mg of ‰opropione in50 mL of the mobile phase. To 20 mL of the solution add25 mL of a solution prepared by dissolving 25 mg of ethylparahydroxybenzoate in 30 mL of acetonitrile and add waterto make 50 mL, and then add the mobile phase to make50 mL. When the procedure is run with 5 mL of this solutionunder the above operating conditions, ‰opropione and ethylparahydroxybenzoate are eluted in this order with the reso-lution between these peaks being not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of ‰opropione is not more than 1.0z.
Containers and storage Containers—Tight containers.
Fosfomycin Calciumホスホマイシンカルシウム
Change the origin/limits of content to read:
Fosfomycin Calcium is the calcium salt of a sub-stance having antibacterial actively produced by thegrowth of Streptomyces fradiae or by the chemicalsynthesis.
It contains not less than 725 mg (potency) and notmore than 805 mg (potency) per mg, calculated on theanhydrous basis. The potency of Fosfomycin Calciumis expressed as mass (potency) of fosfomycin(C3H7O4P: 138.06).
Change the Description to read:
Description Fosfomycin Calcium occurs as a white crystal-line powder.
It is slightly soluble in water, and practically insoluble inmethanol and in ethanol (99.5).
Change the Identiˆcation (2) to read:
Identiˆcation(2) Determine the spectrum of a solution of Fosfomycin
Calcium in heavy water for nuclear magnetic resonancespectroscopy (1 in 300), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound, as directedunder the Nuclear Magnetic Resonance Spectroscopy: itexhibits a double signal at around d 1.5 ppm, a duple doublesignal at around d 2.9 ppm, a multiple signal at around d 3.3ppm, and no signal at around d 1.4 ppm.
Change the Purity (1) to read:
Purity (1) Heavy metals—To 1.0 g of FosfomycinCalcium add 40 mL of 0.25 mol/L acetic acid TS and waterto make 50 mL. Proceed with this solution according toMethod 1, and perform the test. Prepare the control solutionwith 2.0 mL of Standard Lead Solution (not more than 20ppm).
Fosfomycin Sodiumホスホマイシンナトリウム
Change the origin/limits of content to read:
Fosfomycin Sodium is the sodium salt of a sub-stance having antibacterial activity produced by thegrowth of Streptomyces fradiae or by the chemicalsynthesis.
It contains not less than 725 mg (potency) and notmore than 770 mg (potency) per mg, calculated on theanhydrous basis. The potency of Fosfomycin Sodiumis expressed as mass (potency) of fosfomycin(C3H7O4P: 138.06).
Change the Description to read:
Description Fosfomycin Sodium occurs as a white crystal-line powder.
It is very soluble in water, sparingly soluble in methanol,and practically insoluble in ethanol (99.5).
Change the Identiˆcation (2) to read:
Identiˆcation(2) Determine the spectrum of a solution of Fosfomycin
Sodium in heavy water for nuclear magnetic resonancespectroscopy (1 in 300), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound, as directedunder the Nuclear Magnetic Resonance Spectroscopy: itexhibits a double signal at around d 1.5 ppm, a duple doublesignal at around d 2.8 ppm, a multiple signal at around d3.3 ppm, and no signal at around d 1.3 ppm.
17141714 Supplement II, JPXIVO‹cial Monographs for Part I
Furosemideフロセミド
Change the origin/limits of content to read:
Furosemide, when dried, contains not less than98.0z and not more than 101.0z of C12H11ClN2O5S.
Change the Description to read:
Description Furosemide occurs as white, crystals or crys-talline powder.
It is freely soluble in N,N-dimethylformamide, soluble inmethanol, sparingly soluble in ethanol (99.5), slightly solu-ble in acetonitrile and in acetic acid (100), and practicallyinsoluble in water.
It dissolves in dilute sodium hydroxide TS.It is gradually colored by light.Melting point: about 2059C (with decomposition).
Change the Identiˆcation to read:
Identiˆcation (1) Dissolve 25 mg of Furosemide in 10 mLof methanol. To 1 mL of this solution add 10 mL of 2mol/L hydrochloric acid TS. Heat the solution under are‰ux condenser on a water bath for 15 minutes, cool, andadd 18 mL of sodium hydroxide TS to make weakly acidic:the solution responds to the Qualitative Tests for primaryaromatic amines, producing a red to red-purple color.
(2) Determine the absorption spectrum of a solution ofFurosemide in dilute sodium hydroxide TS (1 in 125,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Furosemide ReferenceStandard prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption atthe same wavelengths.
(3) Determine the infrared absorption spectrum ofFurosemide as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or thespectrum of Furosemide Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.
Change the Purity to read:
Purity (1) Clarity and color of solution—Dissolve 0.5 gof Furosemide in 10 mL of a solution of sodium hydroxide(1 in 50): the solution is clear and colorless.
(2) Chloride—Dissolve 2.6 g of Furosemide in 90 mL ofdilute sodium hydroxide TS, add 2 mL of nitric acid, andˆlter. To 25 mL of the ˆltrate add 6 mL of dilute nitric acidand water to make 50 mL, and perform the test using this so-lution as the test solution. Prepare the control solution asfollows: To 0.40 mL of 0.01 mol/L hydrochloric acid VSadd 6 mL of dilute nitric acid and water to make 50 mL (not
more than 0.020z).(3) Sulfate—To 20 mL of the ˆltrate obtained in (2) add
1 mL of dilute hydrochloric acid and water to make 50 mL,and perform the test using this solution as the test solution.Prepare the control solution as follows: To 0.35 mL of 0.005mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acidand water to make 50 mL (not more than 0.030z).
(4) Heavy metals—Proceed with 2.0 g of Furosemideaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 10 ppm).
(5) Related substances—Dissolve 25 mg of Furosemidein 25 mL of the dissolving solution, and use this solution asthe sample solution. Pipet 1 mL of the sample solution, addthe dissolving solution to make exactly 200 mL, and use thissolution as the standard solution. Perform the test with 20mL each of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the area of each peakappeared ahead of the peak of furosemide is not more than2/5 times the peak area of furosemide from the standard so-lution, the area of each peak appeared behind the peak offurosemide is not more than 1/4 times the peak area offurosemide from the standard solution, and the total area ofthese peaks is not more than 2 times the peak area offurosemide from the standard solution.Dissolving solution—To 22 mL of acetic acid (100) add amixture of water and acetonitrile (1:1) to make 1000 mL.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 272 nm).
Column: A stainless steel column 4.6 mm in inside di-ameter and 25 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of water, tetrahydrofuran andacetic acid (100) (70:30:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof furosemide is about 18 minutes.
Time span of measurement: About 2.5 times as long as theretention time of furosemide after the solvent peak.System suitability—
Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the dissolving solution tomake exactly 50 mL. Conˆrm that the peak area offurosemide obtained from 20 mL of this solution is equiva-lent to 3.2 to 4.8z of that obtained from 20 mL of thestandard solution.
System performance: When the procedure is run with 20mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of furosemide is not less than 7000 andnot more than 1.5, respectively.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-
17151715Supplement II, JPXIV O‹cial Monographs for Part I
ing conditions, the relative standard deviation of the peakarea of furosemide is not more than 2.0z.
Change the Assay to read:
Assay Weigh accurately about 0.5 g of Furosemide, previ-ously dried, dissolve in 50 mL of N,N-dimethylformamide,and titrate with 0.1 mol/L sodium hydroxide VS until thecolor of the solution changes from yellow to blue (indicator:3 drops of bromothymol blue TS). Perform a blank determi-nation with a mixture of 50 mL of N,N-dimethylformamideand 15 mL of water, and make any necessary correction
Each mL of 0.1 mol/L sodium hydroxide VS＝33.07 mg of C12H11ClN2O5S
Add the following:
Furosemide Tabletsフロセミド錠
Furosemide Tablets contain not less than 95.0z andnot more than 105.0z of the labeled amount offurosemide (C12H11ClN2O5S: 330.74).
Method of preparation Prepare as directed under Tablets,with Furosemide.
Identiˆcation (1) Shake well a quantity of powderedFurosemide Tablets, equivalent to 0.2 g of Furosemideaccording to the labeled amount, with 40 mL of acetone,and ˆlter. To 0.5 mL of the ˆltrate add 10 mL of 2 mol/Lhydrochloric acid TS, and heat under a re‰ux condenser on awater bath for 15 minutes. After cooling, add 18 mL ofsodium hydroxide TS to make the solution slightly acetic:the solution responds to the Qualitative Tests for primaryaromatic amines, producing a red to red-purple color.
(2) Determine the absorption spectrum of the samplesolution obtained in the Assay as directed under theUltraviolet-visible Spectrophotometry: it exhibits maximabetween 227 nm and 231 nm, between 269 nm and 273 nm,and between 330 nm and 336 nm.
Purity To a quantity of powdered Furosemide Tablets,equivalent to 40 mg of Furosemide according to the labeledamount, add about 30 mL of acetone, shake well, and addacetone to make exactly 50 mL. Centrifuge the solution, add3.0 mL of water to 1.0 mL of the supernatant liquid, cool inice, add 3.0 mL of dilute hydrochloric acid and 0.15 mL ofsodium nitrite TS, shake, and allow to stand for 1 minute.Add 1.0 mL of ammonium amidosulfate TS, shake well,allow to stand for 3 minutes, add 1.0 mL of N,N-diethyl-N?-1-naphthylethylenediamine oxalate TS, shake well, andallow to stand for 5 minutes. Perform the test with this solu-tion as directed under the Ultraviolet-visible Spectrophoto-metry, using a solution prepared in the same manner with1.0 mL of acetone as the blank: the absorbance at 530 nm isnot more than 0.10.
Content uniformity Perform the test according to the fol-lowing method: it meets the requirements of the ContentUniformity Test.
To 1 tablet of Furosemide Tablets add a suitable amountof 0.05 mol/L sodium hydroxide TS, shake to disintegrate,then add 0.05 mol/L sodium hydroxide TS to make exactlyV mL so that each mL contains about 0.4 mg of furosemide(C12H11ClN2O5S). Filter the solution, discard the ˆrst 10 mLof the ˆltrate, pipet the subsequent 2 mL of the ˆltrate, add0.05 mol/L sodium hydroxide TS to make exactly 100 mL,and use this solution as the sample solution. Proceed asdirected in the Assay.
Amount (mg) of furosemide (C12H11ClN2O5S)
＝WS×AT
AS×
V100
WS: Amount (mg) of Furosemide Reference Standard
Dissolution Perform the test with 1 tablet of FurosemideTablets at 50 revolutions per minute according to Method 2under the Dissolution Test, using 900 mL of diluted phos-phate buŠer solution, pH 6.8 (1 in 2) as the dissolution medi-um. Withdraw 20 mL or more of the dissolution medium 15minutes after starting the test for a 20-mg tablet or 30minutes after for a 40-mg tablet, and ˆlter through a mem-brane ˆlter with pore size of not more than 0.45 mm. Discardthe ˆrst 10 mL of the ˆltrate, pipet V mL of the subsequentˆltrate, add diluted phosphate buŠer solution, pH 6.8 (1 in2) to make exactly V? mL so that each mL contains about 10mg of furosemide (C12H11ClN2O5S) according to the labeledamount, and use this solution as the sample solution.Separately, weigh accurately about 20 mg of FurosemideReference Standard, previously dried at 1059C for 4 hours,and dissolve in 5 mL of methanol, and add diluted phos-phate buŠer solution, pH 6.8 (1 in 2) to make exactly 100mL. Pipet 5 mL of this solution, add diluted phosphatebuŠer solution, pH 6.8 (1 in 2) to make exactly 100 mL, anduse this solution as the standard solution. Determine theabsorbances, AT and AS, of the sample solution and thestandard solution at 277 nm as directed under the Ultrav-iolet-visible Spectrophotometry: the dissolution rates for a20-mg tablet in 15 minutes and for a 40-mg tablet in 30minutes are not less than 80z, respectively.
Dissolution rate (z) with respect to the labeled amount offurosemide (C12H11ClN2O5S)
＝WS×AT
AS×
V?V×
1C×45
WS: Amount (mg) of Furosemide Reference StandardC: Labeled amount (mg) of furosemide (C12H11ClN2O5S)
in 1 tablet
Assay Weigh accurately the mass of not less than 20Furosemide Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 40 mg of furosemide(C12H11ClN2O5S), add about 70 mL of 0.05 mol/L sodiumhydroxide TS, shake well, and add 0.05 mol/L sodiumhydroxide TS to make exactly 100 mL. Filter, discard theˆrst 10 mL of the ˆltrate, pipet 2 mL of the subsequent
17161716 Supplement II, JPXIVO‹cial Monographs for Part I
ˆltrate, add 0.05 mol/L sodium hydroxide TS to make ex-actly 100 mL, and use this solution as the sample solution.Separately, weigh accurately about 20 mg of FurosemideReference Standard, previously dried at 1059C for 4 hours,and dissolve in 0.05 mol/L sodium hydroxide TS to makeexactly 50 mL. Pipet 2 mL of this solution, add 0.05 mol/Lsodium hydroxide TS to make exactly 100 mL, and use thissolution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and the standardsolution at 271 nm as directed under the Ultraviolet-visibleSpectrophotometry.
Amount (mg) of furosemide (C12H11ClN2O5S)＝WS×AT
AS
WS: Amount (mg) of Furosemide Reference Standard
Containers and storage Containers—Tight containers.Storage—Light-resistant.
Glutathione, when dried, contains not less than98.0z and not more than 101.0z of C10H17N3O6S.
Description Glutathione occurs as a white crystallinepowder.
It is freely soluble in water, and practically insoluble inethanol (99.5).
Melting point: about 1859C (with decomposition).
Identiˆcation Determine the infrared absorption spectrumof Glutathione, previously dried, as directed in the potassi-um bromide disk method under the Infrared Spectrophoto-metry, and compare the spectrum with the Reference Spec-trum: both spectra exhibit similar intensities of absorption atthe same wave numbers.
Purity (1) Clarity and color of solution—Dissolve 1.0 gof Glutathione in 10 mL of water: the solution is clear andcolorless.
(2) Heavy metals—Proceed with 2.0 g of Glutathioneaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 10 ppm).
(3) Arsenic—Prepare the test solution with 1.0 g of
Glutathione according to Method 1, and perform the test(not more than 2 ppm).
(4) Related substances—Dissolve 50 mg of Glutathionein 100 mL of the mobile phase, and use this solution as thesample solution. Pipet 2 mL of the sample solution, add themobile phase to make exactly 100 mL, and use this solutionas the standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the area of the peak havingthe relative retention time of about 4 with respect toglutathione is not more than 3/4 times the peak area ofglutathione from the standard solution, and the total area ofthe peaks other than the peak of glutathione is not morethan the peak area of glutathione from the standard solu-tion.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 210 nm).
Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about309C.
Mobile phase: Dissolve 6.8 g of potassium dihydrogenphosphate and 2.02 g of sodium 1-heptane sulfonate in 1000mL of water, and adjust the pH to 3.0 with phosphoric acid.To 970 mL of this solution add 30 mL of methanol.
Flow rate: Adjust the ‰ow rate so that the retention timeof glutathione is about 5 minutes.
Time span of measurement: About 6 times as long as theretention time of glutathione after the solvent peak.System suitability—
Test for required detectability: Pipet 10 mL of the stan-dard solution, and add the mobile phase to make exactly 100mL. Conˆrm that the peak area of glutathione obtainedfrom 10 mL of this solution is equivalent to 8 to 12z of thatobtained from 10 mL of the standard solution.
System performance: Dissolve 50 mg of glutathione,10 mg of D-phenylglycine and 50 mg of ascorbic acid in 100mL of water. When the procedure is run with 10 mL of thissolution under the above operating conditions, ascorbicacid, glutathione and D-phenylglycine are eluted in thisorder, and the resolutions between the peaks of ascorbic acidand glutathione and between the peaks of glutathione andD-phenylglycine are not less than 5, respectively.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of glutathione is not more than 1.5z.
Loss on drying Not more than 0.5z (1 g, 1059C, 3 hours).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately about 0.5 g of Glutathione,previously dried, dissolve in 50 mL of a solution of
17171717Supplement II, JPXIV O‹cial Monographs for Part I
metaphosphoric acid (1 in 50), and titrate with 0.05 mol/Liodine VS (indicator: 1 mL of starch TS). Perform a blankdetermination in the same manner, and make any necessarycorrection.
Each mL of 0.05 mol/L iodine VS＝30.73 mg of C10H17N3O6S
Containers and storage Containers—Tight containers.
Heparin Sodiumヘパリンナトリウム
Change the Assay to read:
Assay (i) Substrate solution: Dissolve 15 mg of N-ben-zoyl-L-isoleucyl-L-glutamyl(g-OR)-glycyl-L-arginyl-p-nitroanilide hydrochloride in 20 mL of water.
(ii) Antithrombin III solution: Dissolve human an-tithrombin III in water to make a solution containing 1 Unitper mL.
(iii) Activated blood coagulation factor X solution: Dis-solve bovine activated blood coagulation factor X in waterto make a solution containing 0.426 Units per mL.
(iv) Human normal plasma: Dissolve an amount ofdried human normal plasma powder, equivalent to 1 mL ofhuman normal plasma, in 1 mL of water. Store at 2 – 109Cand use within a week.
(v) BuŠer solution: Dissolve 6.06 g of 2-amino-2-hydroxymethyl-1,3-propanediol in 750 mL of water, adjustthe pH to 8.4 with 1 mol/L hydrochloric acid TS, and addwater to make 1000 mL.
(vi) Reaction stop solution: To 20 mL of acetic acid(100) add water to make 40 mL.
(vii) Heparin standard solution: Dissolve Heparin Sodi-um Reference Standard in isotonic sodium chloride solutionto make a solution containing 10 Units per mL, and use asthe standard stock solution. To 100 mL of the standard stocksolution add the buŠer solution to make exactly 5 mL, anduse this solution as the standard solution. Prepare theheparin standard solutions (1), (2), (3), (4) and (5) byaddition of antithrombin III solution, human normalplasma and the buŠer solution to the standard solution asdirected in the following table.
Heparin standardsolution BuŠer
solution(mL)
AntithrombinIII solution
(mL)
Humannormalplasma(mL)
Standardsolution
(mL)No.Heparin
concentration(Unit/mL)
(1) 0 800 100 100 0
(2) 0.02 700 100 100 100
(3) 0.04 600 100 100 200
(4) 0.06 500 100 100 300
(5) 0.08 400 100 100 400
(viii) Sample solution: Weigh accurately an adequateamount of Heparin Sodium, dissolve in isotonic sodiumchloride solution so that each mL contains about 0.5 Unitsaccording to the labeled amount. To 100 mL of this solutionadd 100 mL of antithrombin III solution, 100 mL of humannormal plasma and 700 mL of the buŠer solution, and usethis solution as the sample solution.
(ix) Procedure: Transfer 400 mL of the sample solutionto a test tube, and warm at 379C for 4 minutes. Add 200 mLof the activated blood coagulation factor X solution, mixwell, warm at 379C for exactly 30 seconds, add 400 mL of thesubstrate solution, previously warmed at 379C, and mixwell. Allow the tube to stand at 379C for exactly 3 minutes,add 600 mL of the reaction stop solution, mix immediately,and determine the absorbance at 405 nm, using the blanksolution prepared by addition of 600 mL of the reaction stopsolution and 600 mL of water to 400 mL of the samplesolution. Proceed the same way with the heparin standardsolution (1), the heparin standard solution (2), the heparinstandard solution (3), the heparin standard solution (4) andthe heparin standard solution (5), and determine their absor-bances.
(x) Calculation: Plot the absorbances of the standard so-lutions on the vertical axis and their heparin concentrationson the horizontal axis to prepare a calibration curve. Deter-mine the heparin concentration, C, of the sample solutionfrom its absorbance by using the curve, and calculateheparin Units per mg of Heparin Sodium from the followingformula.
Units per mg of Heparin Sodium＝C×10×ba
a: Amount of sample (mg)b: Total volume (mL) of isotonic sodium chloride solution
used to dissolve the sample to make the solutioncontaining about 0.5 Units per mL
17181718 Supplement II, JPXIVO‹cial Monographs for Part I
Heparin Sodium Injectionヘパリンナトリウム注射液
Add the following next to Bacterial endotoxins:
Actual volume It meets the requirements of Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Change the Assay to read:
Assay Proceed as directed in the Assay under HeparinSodium, replacing the sample solution indicated in (viii) andthe calculation in (x) with the following.
Sample solution: Measure exactly an adequate portion ofHeparin Sodium Injection according to the labeled Units,dilute it with isotonic sodium chloride solution so that eachmL contains about 0.5 Units. To 100 mL of this solution add100 mL of antithrombin III solution, 100 mL of human nor-mal plasma and 700 mL of the buŠer solution, and use thissolution as the sample solution.
Calculation: Plot the absorbances of the standard solu-tions on the vertical axis and their heparin concentrations onthe horizontal axis to prepare a calibration curve. Determinethe heparin concentration, C, of the sample solution from itsabsorbance by using the curve, and calculate heparin Unitsper mL of Heparin Sodium Injection from the followingformula.
Units per mL of Heparin Sodium Injection＝C×10×ba
a: Amount of sample (mL)b: Total volume (mL) of isotonic sodium chloride solution
used to dilute the sample to make the solution contain-ing about 0.5 Units per mL
Kallidinogenaseカリジノゲナーゼ
Change the Assay to read:
Assay Weigh accurately an appropriate amount of Kal-lidinogenase according to the labeled Units, dissolve in 0.05mol/L phosphate buŠer solution, pH 7.0 to prepare a solu-tion containing about 10 Kallidinogenase Units per mL, anduse this solution as the sample stock solution. Pipet 4 mL ofthe sample stock solution, add exactly 1 mL of trypsin inhi-
bitor TS and 0.05 mol/L phosphate buŠer solution, pH 7.0to make exactly 10 mL, and use this solution as the samplesolution. Pipet 2.5 mL of substrate TS for kallidinogenaseassay (1), previously warmed at 30±0.59C for 5 minutes,place in a 1-cm cell, add exactly 0.5 mL of the sample solu-tion, warmed at 30±0.59C for 5 minutes, and start simul-taneously a chronograph. Perform the test at 30±0.59C asdirected under the Ultraviolet-visible Spectrophotometryusing water as the blank, and determine the absorbances at405 nm, AT2 and AT6, of this solution after allowing to standfor exactly 2 and 6 minutes. Separately, dissolve Kal-lidinogenase Reference Standard in 0.05 mol/L phosphatebuŠer solution, pH 7.0 to make a solutin so that each mLcontains exactly 10 Units, and use this solution as the stan-dard stock solution. Pipet 4 mL of the stock solution, addexactly 1 mL of trypsin inhibitor TS and 0.05 mol/L phos-phate buŠer solution, pH 7.0 to make exactly 10 mL, anduse this solution as the standard solution. Take exactly 0.5mL of the standard solution, perform the test in the samemanner as described for the sample solution, and determinethe absorbances, AS2 and AS6, of the solution after allowingto stand for exactly 2 and 6 minutes. Separately, take exactly1 mL of the trypsin inhibitor TS, and add 0.05 mol/L phos-phate buŠer solution, pH 7.0 to make exactly 10 mL. Pipet0.5 mL of this solution, perform the test in the same manneras described for the sample solution, and determine the ab-sorbances, AO2 and AO6, of the solution after allowing tostand for exactly 2 and 6 minutes.
Units per 1 mg of Kallidinogenase
＝(AT6－AT2)－(AO6－AO2)(AS6－AS2)－(AO6－AO2)
×WS
a×
1b
WS: Amount (Units) of Kallidinogenase Reference Standarda: Volume (mL) of the standard stock solutionb: Amount (mg) of Kallidinogenase in 1 mL of the sample
stock solution
D-MannitolD-マンニトール
Change the Identiˆcation (2) to read:
Identiˆcation(2) Determine the infrared absorption spectrum of D-
Mannitol as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.If any diŠerence appears between the spectra, dissolve 1 g ofD-Mannitol in 3 mL of warm water, then allow to stand at59C for 24 hours or until crystals appear, and ˆlter. Washthe crystals so obtained with a few amount of cold water,dry at 1059C for 4 hours, and perform the test with thecrystals.
17191719Supplement II, JPXIV O‹cial Monographs for Part I
dl-Methylephedrine Hydrochloride, when dried,contains not less than 99.0z and not more than101.0z of C11H17NO.HCl.
Change the Description to read:
Description dl-Methylephedrine Hydrochloride occurs ascolorless crystals or a white, crystalline powder.
It is freely soluble in water, sparingly soluble in ethanol(99.5), slightly soluble in acetic acid (100), and practicallyinsoluble in acetic anhydride.
A solution of dl-Methylephedrine Hydrochloride (1 in 20)shows no optical rotation.
Change the Identiˆcation to read:
Identiˆcation (1) Determine the absorption spectrum ofa solution of dl-Methylephedrine Hydrochloride (1 in 2000)as directed under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.
(2) Determine the infrared absorption spectrum of dl-Methylephedrine Hydrochloride, previously dried, as direct-ed in the potassium chloride disk method under the InfraredSpectrophotometry, and compare the spectrum with the
Reference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wave numbers.
(3) A solution of dl-Methylephedrine Hydrochloride(1 in 10) responds to the Qualitative Tests for chloride.
Add the following next to Identiˆcation:
pH The pH of a solution prepared by dissolving 1.0 g ofdl-Methylephedrine Hydrochloride in 20 mL of water is be-tween 4.5 and 6.0.
Change the Purity to read:
Purity (1) Clarity and color of solution—Dissolve 1.0 gof dl-Methylephedrine Hydrochloride in 10 mL of water: thesolution is clear and colorless.
(2) Heavy metals—Proceed with 1.0 g of dl-Methylephe-drine Hydrochloride according to Method 4, and performthe test. Prepare the control solution with 1.0 mL of Stan-dard Lead Solution (not more than 10 ppm).
(3) Related substances—Dissolve 50 mg of dl-Methylephedrine Hydrochloride in 20 mL of water, and usethis solution as the sample solution. Pipet 1 mL of thesample solution, add water to make exactly 100 mL, and usethis solution as the standard solution. Perform the test withexactly 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the totalarea of the peaks other than the peak of methylephedrine isnot more than the peak area of methylephedrine from thestandard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 257 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 13.6 g of potassium dihydrogenphosphate and 3 g of sodium 1-heptane sulfonate in 1000mL of water, and adjust the pH to 2.5 with phosphoric acid.To 900 mL of this solution add 200 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof methylephedrine is about 10 minutes.
Time span of measurement: About 2 times as long as theretention time of methylephedrine after the solvent peak.System suitability—
Test for required detectability: To exactly 2 mL of thestandard solution add water to make exactly 20 mL. Con-ˆrm that the peak area of methylephedrine obtained from 20mL of this solution is equivalent to 7 to 13z of that ofmethylephedrine obtained from 20 mL of the standard solu-tion.
System performance: Dissolve 50 mg of dl-Methylephe-drine Hydrochloride and 0.4 mg of methyl parahydroxyben-zoate in 50 mL of water. When the procedure is run with
17201720 Supplement II, JPXIVO‹cial Monographs for Part I
20 mL of this solution under the above operating conditions,methylephedrine and methyl parahydroxybenzoate areeluted in this order with the resolution between these peaksbeing not less than 3.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of methylephedrine is not more than 2.0z.
Identiˆcation Determine the absorption spectrum of asolution of 10z dl-Methylephedrine Hydrochloride Powder(1 in 200) as directed under the Ultraviolet-visible Spec-trophotometry: it exhibits maxima between 250 nm and253 nm, between 255 nm and 259 nm, and between 261 nmand 264 nm.
Change the Assay to read:
Assay Weigh accurately about 0.5 g of 10z dl-Methylephedrine Hydrochloride Powder, add exactly 4 mLof the internal standard solution and 25 mL of water, shakevigorously for 20 minutes to dissolve, add water to make50 mL, ˆlter through a membrane ˆlter with pore size of0.45 mm, if necessary, discard the ˆrst 10 mL of the ˆltrate,and use the subsequent ˆltrate as the sample solution.Separately, weigh accurately about 50 mg of dl-methylephe-drine hydrochloride for assay, previously dried at 1059C for3 hours, add exactly 4 mL of the internal standard solutionand water to make 50 mL, and use this solution as thestandard solution. Perform the test with 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and determine the ratios of the peak area, QT
and QS, of methylephedrine to that of the internal standard.
Amount (mg) of dl-methylephedrine hydrochloride(C11H17NO.HCl)
＝WS×QT
QS
WS: Amount (mg) of dl-methylephedrine hydrochloridefor assay
Internal standard solution—A solution of methyl para-hydroxybenzoate in acetonitrile (1 in 10,000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 257 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-
silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 13.6 g of potassium dihydrogenphosphate and 3 g of sodium 1-heptane sulfonate in 1000mL of water, and adjust the pH to 2.5 with phosphoric acid.To 900 mL of this solution add 200 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof methylephedrine is about 10 minutes.System suitability—
System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, methylephedrine and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 3.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of methylephedrine to that of the internalstandard is not more than 1.0z.
Methylprednisolone Succinate, when dried, containsnot less than 97.0z and not more than 103.0z ofC26H34O8.
Description Methylprednisolone Succinate occurs as awhite, crystals or crystalline powder.
It is soluble in methanol, sparingly soluble in ethanol (95),and practically insoluble in water.
Melting point: about 2359C (with decomposition).
Identiˆcation (1) Determine the absorption spectrum ofa solution of Methylprednisolone Succinate in methanol(1 in 50,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum or the spectrum of a solution of Methyl-prednisolone Succinate Reference Standard prepared in thesame manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum of
17211721Supplement II, JPXIV O‹cial Monographs for Part I
Methylprednisolone Succinate, previously dried, as directedin the potassium bromide disk method under the InfraredSpectrophotometry, and compare the spectrum with theReference Spectrum or the spectrum of previously driedMethylprednisolone Succinate Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers. In case when some diŠerences are found be-tween the spectra, repeat the test with residues obtained bydissolving these substances in ethanol (95), evaporating todryness, and drying.
Purity (1) Heavy metals—Proceed with 1.0 g of Methyl-prednisolone Succinate according to Method 4, and performthe test. Prepare the control solution with 1.0 mL ofStandard Lead Solution (not more than 10 ppm).
(2) Arsenic—Prepare the test solution with 2.0 g ofMethylprednisolone Succinate according to Method 3, andperform the test (not more than 1 ppm).
(3) Related substances—Dissolve 15 mg of Methylpred-nisolone Succinate in 5 mL of methanol, add a mixture of0.05 mol/L phosphate buŠer solution, pH 3.5 and acetoni-trile (1:1) to make 50 mL, and use this solution as the samplesolution. Pipet 1 mL of the sample solution, add the mixtureof 0.05 mol/L phosphate buŠer solution, pH 3.5 andacetonitrile (1:1) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 5mL each of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the area of the peaks otherthan the peak of methylprednisolone succinate is not morethan 1/2 of the peak area of methylprednisolone succinatefrom the standard solution, and the total area of the peaksother than the peak of methylprednisolone succinate is notmore than the peak area of methylprednisolone succinatefrom the standard solution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 3 times as long as theretention time of methylprednisolone succinate.System suitability—
Test for required detectability: Pipet 1 mL of the standardsolution, and add the mixture of 0.05 mol/L phosphatebuŠer solution, pH 3.5 and acetonitrile (1:1) to make exactly10 mL. Conˆrm that the peak area of methylprednisolonesuccinate obtained from 5 mL of this solution is equivalent to7 to 13z of that obtained from 5 mL of the standard solu-tion.
System performance: Proceed as directed in the Systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of methylprednisolone succinate is not more than
1.0z.
Loss on drying Not more than 1.0z (1 g, 1059C, 3 hours).
Residue on ignition Not more than 0.2z (0.5 g).
Assay Weigh accurately about 15 mg each of Methylpred-nisolone Succinate and Methylprednisolone SuccinateReference Standard, previously dried, dissolve separately in5 mL of methanol, and add the mixture of 0.05 mol/L phos-phate buŠer solution, pH 3.5 and acetonitrile (1:1) to makeexactly 50 mL. Pipet 5 mL each of these solutions, addexactly 5 mL of the internal standard solution, and use thesesolutions as the sample solution and the standard solution,respectively. Perform the test with 5 mL each of the samplesolution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and determine the ratios, QT and QS, of the peak areaof methylprednisolone succinate to that of the internalstandard.
Amount (mg) of C26H34O8＝WS×QT
QS
WS: Amount (mg) of Methylprednisolone SuccinateReference Standard
Internal standard solution—A solution of ethyl parahydrox-ybenzoate in a mixture of 0.05 mol/L phosphate buŠer solu-tion, pH 3.5 and acetonitrile (1:1) (3 in 20,000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: To 1000 mL of 0.05 mol/L potassium di-hydrogen phosphate TS add a suitable amount of 0.05mol/L disodium hydrogen phosphate TS to make a solutionhaving pH 5.5. To 640 mL of this solution add 360 mL ofacetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof methylprednisolone succinate is about 6 minutes.System suitability—
System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, methylprednisolone succinate and the internal stan-dard are eluted in this order with the resolution betweenthese peaks being not less than 6.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of methylprednisolone succinate to that of theinternal standard is not more than 1.0z.
Containers and storage Containers—Tight containers.
17221722 Supplement II, JPXIVO‹cial Monographs for Part I
Methyltestosteroneメチルテストステロン
Change the origin/limits of content to read:
Methyltestosterone, when dried, contains not lessthan 98.0z and not more than 102.0z of C20H30O2.
Change the Description to read:
Description Methyltestosterone occurs as white to paleyellow, crystals or crystalline powder.
It is freely soluble in methanol and in ethanol (95), andpractically insoluble in water.
Change the Identiˆcation to read:
Identiˆcation(1) Determine the absorption spectrum of a solution of
Methyltestosterone in ethanol (95) (1 in 100,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of a solution of Methyltestosterone Reference Stan-dard prepared in the same manner as the sample solution:both spectra exhibit similar intensities of absorption at thesame wavelengths.
(2) Determine the infrared absorption spectrum ofMethyltestosterone, previously dried, as directed in thepotassium bromide disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of dried MethyltestosteroneReference Standard: both spectra exhibit similar intensitiesof absorption at the same wave numbers.
Change the Assay to read:
Assay Weigh accurately about 20 mg each ofMethyltestosterone and Methyltestosterone ReferenceStandard, previously dried in a desiccator (in vacuum, phos-phorus (V) oxide) for 10 hours, dissolve each in methanol tomake exactly 200 mL. Pipet 5 mL each of these solutions,add exactly 5 mL of the internal standard solution, addmethanol to make 50 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of methyltestosterone tothat of the internal standard.
Amount (mg) of C20H30O2
＝WS×QT
QS
WS: Amount (mg) of Methyltestosterone ReferenceStandard
Internal standard solution—A solution of propyl para-hydroxybenzoate in methanol (1 in 10,000).
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 241 nm).Column: A stainless steel column 6 mm in inside diameter
and 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about359C.
Mobile phase: A mixture of acetonitrile and water (11:9).Flow rate: Adjust the ‰ow rate so that the retention time
of methyltestosterone is about 10 minutes.System suitability—
System performance: When the procedure is run with 10mL of the standard solution under the above operatingconditions, the internal standard and methyltestosterone areeluted in this order with the resolution between these peaksbeing not less than 9.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the aboveoperating conditions, the relative standard deviation of theratios of the peak area of methyltestosterone to that of theinternal standard is not more than 1.0z.
Methyltestosterone Tabletsメチルテストステロン錠
Change the origin/limits of content to read:
Methyltestosterone Tablets contain not less than90.0z and not more than 110.0z of the labeledamount of methyltestosterone (C20H30O2: 302.45).
Change the Identiˆcation to read:
Identiˆcation To a portion of powdered Methyltestoster-one Tablets, equivalent to 0.01 g of Methyltestosteroneaccording to the labeled amount, add 50 mL of acetone,shake for 30 minutes, and ˆlter. Evaporate the ˆltrate todryness, dissolve the residue in 10 mL of acetone, and usethis solution as the sample solution. Separately, dissolve0.01 g of Methyltestosterone Reference Standard in 10 mLof acetone, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of chloroform and ethanol (95) (9:1) to a distance ofabout 12 cm, and air-dry the plate. Spray evenly dilute sul-furic acid on the plate, and heat at 1109C for 10 minutes: thespot from the sample solution and the standard solutionshow the same Rf value.
Change the Assay to read:
Assay Weigh accurately the mass of not less than 20Methyltestosterone Tablets, and powder. Weigh accurately aportion of the powder, equivalent to about 25 mg of
17231723Supplement II, JPXIV O‹cial Monographs for Part I
methyltestosterone (C20H30O2), add about 70 mL ofmethanol, shake for 30 minutes, and add methanol to makeexactly 100 mL. Pipet 2 mL of this solution, add exactly 5mL of the internal standard solution and methanol to make50 mL, ˆlter through a membrane ˆlter (not exceeding 0.45mm in pore size), and use the ˆltrate as the sample solution.Separately, weigh accurately about 20 mg ofMethyltestosterone Reference Standard, previously dried ina desiccator (in vacuum, phosphorus (V) oxide) for 10hours, dissolve in methanol to make exactly 200 mL. Pipet 5mL of this solution, add exactly 5 mL of the internal stan-dard solution, add methanol to make 50 mL, and use this so-lution as the standard solution. Perform the test with 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of methyltestosterone to that of the internalstandard.
Amount (mg) of methyltestosterone (C20H30O2)
＝WS×QT
QS×
52
WS: Amount (mg) of Methyltestosterone ReferenceStandard
Internal standard solution—A solution of propyl para-hydroxybenzoate in methanol (1 in 10,000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 241 nm).
Column: A stainless steel column 6 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about359C.
Mobile phase: A mixture of acetonitrile and water (11:9).Flow rate: Adjust the ‰ow rate so that the retention time
of methyltestosterone is about 10 minutes.System suitability—
System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and methyltestosterone areeluted in this order with the resolution between these peaksbeing not less than 9.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of methyltestosterone to that of the internalstandard is not more than 1.0z.
Add the following:
Metoclopramide Tabletsメトクロプラミド錠
Metoclopramide Tablets contain not less than95.0z and not more than 105.0z of the labeledamount of metoclopramide (C14H22ClN3O2: 299.80).
Method of preparation Prepare as directed under Tablets,with Metoclopramide.
Identiˆcation (1) To a quantity of powderedMetoclopramide Tablets, equivalent to 50 mg ofMetoclopramide according to the labeled amount, add 15mL of 0.5 mol/L hydrochloric acid TS, and heat in a waterbath at 709C for 15 minutes while frequent shaking. Aftercooling, centrifuge for 10 minutes, and to 5 mL of the super-natant liquid add 1 mL of 4-dimethylaminobenzaldehyde-hydrochloric acid TS: a yellow color develops.
(2) Determine the absorption spectrum of the samplesolution obtained in the Assay as directed under theUltraviolet-visible Spectrophotometry: it exhibits maximabetween 270 nm and 274 nm, and between 306 nm and310 nm.
Content uniformity Perform the test according to thefollowing method: it meets the requirements of the ContentUniformity Test.
To 1 tablet of Metoclopramide Tablets add 10 mL of 0.1mol/L hydrochloric acid TS, disperse the particles with theaid of ultrasonic waves, then add 0.1 mol/L hydrochloricacid TS to make exactly 25 mL, and centrifuge for 10minutes. Pipet 4 mL of the supernatant liquid, add0.1 mol/L hydrochloric acid TS to make exactly V mL of asolution so that each mL contains about 12 mg ofmetoclopramide (C14H22ClN3O2), and use this solution as thesample solution. Separately, weigh accurately about 80 mgof metoclopramide for assay, previously dried at 1059C for3 hours, and dissolve in 0.1 mol/L hydrochloric acid TS tomake exactly 500 mL. Pipet 4 mL of this solution, add 0.1mol/L hydrochloric acid TS to make exactly 50 mL, and usethis solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and the standardsolution at 308 nm as directed under the Ultraviolet-visibleSpectrophotometry.
Amount (mg) of metoclopramide (C14H22ClN3O2)
＝WS×AT
AS×
V1000
WS: Amount (mg) of metoclopramide for assay
Dissolution Perform the test with 1 tablet ofMetoclopramide Tablets at 50 revolutions per minute ac-cording to Method 2 under the Dissolution Test, using 900mL of diluted phosphate buŠer solution, pH 6.8 (1 in 2) asthe dissolution medium. Withdraw 20 mL or more of thedissolution medium 45 minutes after starting the test for a3.84-mg tablet or 15 minutes after starting the test for a
17241724 Supplement II, JPXIVO‹cial Monographs for Part I
7.67-mg tablet, and ˆlter through a membrane ˆlter withpore size of not more than 0.5 mm. Discard the ˆrst 10 mL ofthe ˆltrate, pipet V mL of the subsequent ˆltrate, add dilut-ed phosphate buŠer solution, pH 6.8 (1 in 2) to make exactlyV? mL so that each mL contains about 4 mg ofmetoclopramide (C14H22ClN3O2) according to the labeledamount, and use this solution as the sample solution.Separately, weigh accurately about 20 mg ofmetoclopramide for assay, previously dried at 1059C for 3hours, and dissolve in diluted phosphate buŠer solution, pH6.8 (1 in 2) to make exactly 100 mL. Pipet 2 mL of this solu-tion, add diluted phosphate buŠer solution, pH 6.8 (1 in 2)to make exactly 100 mL, and use this solution as the stan-dard solution. Perform the test with exactly 50 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine the peak areas, AT and AS, ofmetoclopramide. The dissolution rates for a 3.84-mg tabletin 45 minutes and for a 7.67-mg tablet in 15 minutes are notless than 80z and not less than 85z, respectively.
Dissolution rate (z) with respect to the labeled amount ofmetoclopramide (C14H22ClN3O2)
＝WS×AT
AS×
V?V×
1C×18
WS: Amount (mg) of metoclopramide for assayC: Labeled amount (mg) of metoclopramide
(C14H22ClN3O2) in 1 tablet
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 275 nm).Column: A stainless steel column 4 mm in inside diameter
and 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: Dissolve 0.79 g of sodium lauryl sulfate in550 mL of water, and add 450 mL of acetonitrile and 0.3 mLof acetic acid (100).
Flow rate: Adjust the ‰ow rate so that the retention timeof metoclopramide is about 5 minutes.System suitability—
System performance: When the procedure is run with50 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of metoclopramide are not less than3000 and not more than 1.5, respectively.
System repeatability: When the test is repeated 6 timeswith 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of metoclopramide is not more than 1.0z.
Assay Weigh accurately not less than 20 MetoclopramideTablets, and powder. Weigh accurately a portion of thepowder, equivalent to about 75 mg of metoclopramide(C14H22ClN3O2), add 300 mL of 0.1 mol/L hydrochloric acidTS, shake for 1 hour, and add 0.1 mol/L hydrochloric acidTS to make exactly 500 mL. Centrifuge for 10 minutes, pipet
4 mL of the supernatant liquid, add 0.1 mol/hydrochloricacid TS to make exactly 50 mL, and use this solution as thesample solution. Separately, weigh accurately about 80 mgof metoclopramide for assay, previously dried at 1059C for3 hours, and dissolve in 0.1 mol/L hydrochloric acid TS tomake exactly 500 mL. Pipet 4 mL of this solution, add 0.1mol/hydrochloric acid TS to make exactly 50 mL, and usethis solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and the standardsolution at 308 nm as directed under the Ultraviolet-visibleSpectrophotometry.
Amount (mg) of metoclopramide (C14H22ClN3O2)
＝WS×AT
AS
WS: Amount (mg) of metoclopramide for assay
Containers and storage Containers—Tight containers.
Nicorandil contains not less than 98.5z and notmore than 101.0z of C8H9N3O4, calculated on theanhydrous basis.
Description Nicorandil occurs as white crystals.It is freely soluble in methanol, in ethanol (99.5) and in
acetic acid (100), soluble in acetic anhydride, and sparinglysoluble in water.
Melting point: about 929C (with decomposition).
Identiˆcation Determine the infrared absorption spectrumof Nicorandil as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.
Purity (1) Sulfate—Dissolve 2.0 g of Nicorandil in 20 mLof dilute ethanol, add 1 mL of dilute hydrochloric acid andwater to make 50 mL, and perform the test using this solu-tion as the test solution. Prepare the control solution with0.40 mL of 0.005 mol/L sulfuric acid VS, 20 mL of diluteethanol and 1 mL of dilute hydrochloric acid, and dilutewith water to make 50 mL (not more than 0.010z).
(2) Heavy metals—Proceed with 2.0 g of Nicorandilaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 10 ppm).
17251725Supplement II, JPXIV O‹cial Monographs for Part I
(3) Related substances—Dissolve 20 mg of Nicorandil in10 mL of the mobile phase, and use this solution as thesample solution. Perform the test with 10 mL of the samplesolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the peakarea of N-(2-hydroxyethyl)isonicotinamide nitric ester,having the relative retention time of about 0.86 with respectto nicorandil, is not more than 0.5z of the peak area ofnicorandil, the area of all other peaks is less than 0.1z, andthe sum area of the peaks other than nicorandil and N-(2-hydroxyethyl)isonicotinamide nitric ester is not more than0.25z of the total peak area.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4 mm in inside diameterand 25 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of water, tetrahydrofuran,triethylamine and tri‰uoroacetic acid (982:10:5:3).
Flow rate: Adjust the ‰ow rate so that the retention timeof nicorandil is about 18 minutes.
Time span of measurement: About 3 times as long as theretention time of nicorandil after the solvent peak.System suitability—
Test for required detectability: Measure exactly 1 mL ofthe sample solution, add the mobile phase to make exactly500 mL, and use this solution as the solution for systemsuitability test. Pipet 1 mL of the solution for systemsuitability test, and add the mobile phase to make exactly20 mL. Conˆrm that the peak area of nicorandil obtainedwith 10 mL of this solution is equivalent to 2 to 8z of thatwith 10 mL of the solution for system suitability test.
System performance: Dissolve 10 mg of N-(2-hydrox-yethyl)isonicotinamide nitric ester in the mobile phase tomake 100 mL. To 1 mL of this solution add 10 mL of thesample solution. When the procedure is run with this solu-tion under the above operating conditions, N-(2-hydrox-yethyl)isonicotinamide nitric ester and nicorandil are elutedin this order with the resolution between these peaks beingnot less than 3.0.
System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of nicorandil is not more than 1.5z.
Water Not more than 0.1z (2 g, volumetric titration,direct titration).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately about 0.3 g of Nicorandil, dis-solve in 30 mL of a mixture of acetic anhydride and aceticacid (100) (7:3), and titrate with 0.1 mol/L perchloric acidVS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correc-
tion.
Each mL of 0.1 mol/L perchloric acid VS＝21.12 mg of C8H9N3O4
Containers and storage Containers—Tight containers.Storage—At a temperature between 29C and 89C.
Nilvadipine contains not less than 98.0z and notmore than 102.0z of C19H19N3O6.
Description Nilvadipine occurs as a yellow crystallinepowder.
It is freely soluble in acetonitrile, soluble in methanol,sparingly soluble in ethanol (99.5), and practically insolublein water.
A solution of Nilvadipine in acetonitrile (1 in 20) shows nooptical rotation.
Identiˆcation (1) Determine the absorption spectrum ofa solution of Nilvadipine in ethanol (99.5) (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Nilvadipine ReferenceStandard prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption atthe same wavelengths.
(2) Determine the infrared absorption spectrum ofNilvadipine as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or thespectrum of Nilvadipine Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.
Melting point 167 – 1719C
Purity (1) Heavy metals—Proceed with 2.0 g of Nilvadi-pine according to Method 2, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 10 ppm).
(2) Related substances—Dissolve 20 mg of Nilvadipinein 20 mL of acetonitrile, and use this solution as the sample
17261726 Supplement II, JPXIVO‹cial Monographs for Part I
solution. Perform the test with 5 mL of the sample solutionas directed under the Liquid Chromatography according tothe following conditions. Determine each peak area by theautomatic integration method, and calculate the amount ofthem by the area percentage method: the amount of eachrelated substance is not more than 0.3z, and the total ofthem is not more than 0.5z.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 240 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of phosphate buŠer solution,pH 7.4, methanol and acetonitrile (32:27:18).
Flow rate: Adjust the ‰ow rate so that the retention timeof nilvadipine is about 12 minutes.
Time span of measurement: About 2.5 times as long as theretention time of nilvadipine after the solvent peak.System suitability—
Test for required detectability: Pipet 1 mL of the samplesolution, add acetonitrile to make exactly 100 mL, and usethis solution as the solution for system suitability test. Pipet1 mL of the solution for system suitability test, and addacetonitrile to make exactly 10 mL. Conˆrm that the peakarea of nilvadipine obtained from 5 mL of this solution isequivalent to 7 to 13z of that obtained from 5 mL of thesolution for system suitability test.
System performance: When the procedure is run with 5 mLof the solution for system suitability test under the aboveoperating conditions, the number of theoretical plates andthe symmetry factor of the peak of nilvadipine is not lessthan 3300 and not more than 1.3, respectively.
System repeatability: Pipet 1 mL of the solution forsystem suitability test, and add acetonitrile to make exactly10 mL. When the test is repeated 6 times with 5 mL of thissolution under the above operating conditions, the relativestandard deviation of the peak area of nilvadipine is notmore than 1.5z.
Loss on drying Not more than 0.1z (1 g, 1059C, 2 hours).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately about 25 mg each of Nilvadipineand Nilvadipine Reference Standard, dissolve in methanol tomake exactly 25 mL. Pipet 10 mL each of these solutions,add exactly 20 mL of the internal standard solution, 20 mLof water and methanol to make 100 mL, and use these solu-tions as the sample solution and the standard solution, re-spectively. Perform the test with 5 mL each of the sample so-lution and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area of nilvadi-pine to that of the internal standard.
Amount (mg) of C19H19N3O6＝WS×QT
QS
WS: Amount (mg) of Nilvadipine Reference Standard
Internal standard solution—A solution of acenaphthene inmethanol (1 in 200).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: Dissolve 2.5 g of diammonium hydrogenphosphate in 1000 mL of water, add 10 mL of tetrabutylam-monium hydoxide TS, adjust the pH to 7.0 with dilutedphosphoric acid (1 in 10), and add 900 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof nilvadipine is about 12 minutes.System suitability—
System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, nilvadipine and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 8.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of nilvadipine to that of the internal standardis not more than 1.0z.
Containers and storage Containers—Well-closed contain-ers.
Add the following:
Nilvadipine Tabletsニルバジピン錠
Nilvadipine Tablets contain not less than 93.0z andnot more than 107.0z of the labeled amount ofnilvadipine (C19H19N3O6: 385.37).
Method of preparation Prepare as directed under Tablets,with Nilvadipine.
Identiˆcation To a quantity of powdered NilvadipineTablets, equivalent to 1 mg of Nilvadipine according to thelabeled amount, add 100 mL of ethanol (99.5), shake for 10minutes, centrifuge, and use the supernatant liquid as thesample solution. Determine the absorption spectrum of thesample solution as directed under the Ultraviolet-visibleSpectrophotometry: it exhibits a maximum between 239 nmand 243 nm and a maximum having a broad-ranging absorp-tion between 371 nm and 381 nm.
17271727Supplement II, JPXIV O‹cial Monographs for Part I
Content uniformity Perform the test according to the fol-lowing method: it meets the requirements of the ContentUniformity Test.
To 1 tablet of Nilvadipine Tablets add V mL of a mixtureof acetonitrile and water (7:3) so that each mL of the solu-tion contains about 200 mg of nilvadipine (C19H19N3O6)according to the labeled amount, add exactly V mL of theinternal standard solution obtained in the Assay, and dis-perse the particles with the aid of ultrasonic waves. Cen-trifuge for 10 minutes, and use the supernatant liquid as thesample solution. Separately, weigh accurately about 20 mgof Nilvadipine Reference Standard, dissolve in the mixtureof acetonitrile and water (7:3) to make exactly 20 mL. Pipet5 mL of this solution, add exactly 25 mL of the internalstandard solution obtained in the Assay and the mixture ofacetonitrile and water (7:3) to make 50 mL, and use thissolution as the standard solution. Proceed as directed in theAssay.
Amount (mg) of nilvadipine (C19H19N3O6)
＝WS×QT
QS×
V100
WS: Amount (mg) of Nilvadipine Reference Standard
Dissolution Perform the test with 1 tablet of NilvadipineTablets at 50 revolutions per minute according to Method 2under the Dissolution Test, using 900 mL of water as thedissolution medium. Withdraw 20 mL or more of the disso-lution medium 30 minutes after starting the test, and ˆlterthrough a membrane ˆlter with pore size of not more than0.5 mm. Discard the ˆrst 10 mL of the ˆltrate, pipet 10 mL ofthe subsequent ˆltrate, add exactly 1 mL of methanol, anduse this solution as the sample solution. Separately, weighaccurately an amount of Nilvadipine Reference Standard,equivalent to 10 times the labeled amount of NilvadipineTablets, and dissolve in methanol to make exactly 50 mL.Pipet 5 mL of this solution, and add methanol to makeexactly 100 mL. Pipet 1 mL of this solution, add exactly10 mL of water, and use this solution as the standardsolution. Perform the test with exactly 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine the peak areas, AT and AS, of nilvadi-pine: the dissolution rate in 30 minutes is not less than 85z.
Dissolution rate (z) with respect to the labeled amount ofnilvadipine (C19H19N3O6)
＝WS×AT
AS×
1C×9
WS: Amount (mg) of Nilvadipine Reference StandardC: Labeled amount (mg) of nilvadipine (C19H19N3O6) in 1
tablet
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 242 nm).Column: A stainless steel column 4 mm in inside diameter
and 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of phosphate buŠer solution, pH7.4, methanol and acetonitrile (7:7:6)
Flow rate: Adjust the ‰ow rate so that the retention timeof nilvadipineis about 5 minutes.System suitability—
System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetryfactor of the peak of nilvadipine are not less than 2000 andnot more than 1.5, respectively.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of nilvadipine is not more than 1.5z.
Assay Weigh accurately not less than 20 NilvadipineTablets, and powder. Weigh accurately an amount of thepowder, equivalent to about 5 mg of nilvadipine(C19H19N3O6), add 10 mL of a mixture of acetonitrile andwater (7:3) and exactly 25 mL of the internal standard solu-tion, shake for 15 minutes, and add the mixture of acetoni-trile and water (7:3) to make 50 mL. Centrifuge, and use thesupernatant liquid as the sample solution. Separately, weighaccurately about 20 mg of Nilvadipine Reference Standard,dissolve in the mixture of acetonitrile and water (7:3) tomake exactly 20 mL. Pipet 5 mL of this solution, add exactly25 mL of the internal standard solution and the mixture ofacetonitrile and water (7:3) to make 50 mL, and use thissolution as the standard solution. Perform the test with 5 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of nilvadipine to that of the internal stan-dard.
Amount (mg) of nilvadipine (C19H19N3O6)＝WS×QT
QS×
14
WS: Amount (mg) of Nilvadipine Reference Standard
Internal standard solution—A solution of acenaphthene inacetonitrile (1 in 500).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: Dissolve 2.5 g of diammonium hydrogenphosphate in 1000 mL of water, add 10 mL of tetrabutylam-monium hydoxide TS, adjust the pH to 7.0 with dilutedphosphoric acid (1 in 10), and add 900 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof nilvadipine is about 12 minutes.System suitability—
System performance: When the procedure is run with 5 mL
17281728 Supplement II, JPXIVO‹cial Monographs for Part I
of the standard solution under the above operating condi-tions, nilvadipine and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 8.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of nilvadipine to that of the internal standardis not more than 1.0z.
Containers and storage Containers—Well-closed contain-ers.
Add the following:
Oxytocinオキシトシン
C43H66N12O12S2: 1007.19[50-56-6]
Oxytocin is a synthetic peptide having the propertyof causing the contraction of uterine smooth muscle.
It contains not less than 540 oxytocin Units and notmore than 600 oxytocin Units per mg, calculated onthe dehydrated and de-acetic acid basis.
Description Oxytocin occurs as a white powder.It is very soluble in water, and freely soluble in ethanol
(99.5).It dissolves in hydrochloric acid TS.The pH of a solution prepared by dissolving 0.10 g of
Oxytocin in 10 mL of freshly boiled and cooled water is be-tween 4.0 and 6.0.
It is hygroscopic.
Identiˆcation Determine the absorption spectrum of asolution of Oxytocin (1 in 2000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.
Constituent amino acids Put about 1 mg of Oxytocin in atest tube for hydrolysis, add 6 mol/L hydrochloric acid TSto dissolve, replace the air in the tube with Nitrogen, seal thetube under reduced pressure, and heat at 110 to 1159C for16 hours. After cooling, open the tube, evaporate thehydrolyzate to dryness under reduced pressure, add 2 mL of0.02 mol/L hydrochloric acid TS to dissolve the residue, anduse this solution as the sample solution. Separately, weighaccurately about 27 mg of L-aspartic acid, about 24 mg ofL-threonine, about 21 mg of L-serine, about 29 mg ofL-glutamic acid, about 23 mg of L-proline, about 15 mg ofglycine, about 18 mg of L-alanine, about 23 mg of L-valine,about 48 mg of L-cystine, about 30 mg of methionine, about26 mg of L-isoleucine, about 26 mg of L-leucine, about36 mg of L-tyrosine, about 33 mg of phenylalanine, about
37 mg of L-lysine hydrochloride, about 42 mg of L-histidinehydrochloride monohydrate and about 42 mg of L-argininehydrochloride, dissolve them in 10 mL of 1 mol/Lhydrochloric acid TS, and add water to make exactly100 mL. Pipet 5 mL of this solution, add water to make ex-actly 20 mL, and use this solution as the standard solution.Perform the test with exactly 20 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, andcalculate the respective molar ratios with respect to leucine:0.95 – 1.05 for aspartic acid, 0.95 – 1.05 for glutamic acid,0.95 – 1.05 for proline, 0.95 – 1.05 for glycine, 0.80 – 1.10for isoleucine, 0.80 – 1.05 for tyrosine and 0.80 – 1.05 forcystine, and not more than 0.01 each for others.Operating conditions—
Detector: A visible spectrophotometer (wavelength: 440nm and 570 nm).Column: A stainless steel column 4.6 mm in inside diameterand 8 cm in length, packed with strongly acidic ion-exchangeresin for liquid chromatography (sodium type) composedwith a sulfonated polystyrene copolymer (3 mm in particlediameter).
Column temperature: A constant temperature of about579C.
Chemical reaction bath temperature: A constant tempera-ture of about 1309C.
Color developing time: About 1 minute.Mobile phase: Prepare mobile phases A, B and C accord-
ing to the following table.
Mobile phase A B C
Citric acid mono-hydrate 19.80 g 22.00 g 6.10 g
Trisodium citratedihydrate 6.19 g 7.74 g 26.67 g
Sodium chloride 5.66 g 7.07 g 54.35 gEthanol (99.5) 260.0 mL 20.0 mL —Benzyl alcohol — — 5.0 mLThiodiglycol 5.0 mL 5.0 mL —Lauromacrogolsolution (1 in 4) 4.0 mL 4.0 mL 4.0 mL
Capryric acid 0.1 mL 0.1 mL 0.1 mLWater a su‹cient
amounta su‹cient
amounta su‹cient
amount
Total amount 2000 mL 1000 mL 1000 mL
pH 3.3 3.2 4.9
Flowing of the mobile phase: Control the gradient by mix-ing the mobile phases A, B and C as directed in the followingtable.
17291729Supplement II, JPXIV O‹cial Monographs for Part I
Time afterinjection ofsample (min)
MobilephaseA (z)
MobilephaseB (z)
MobilephaseC (z)
0 – 9 100 0 09 – 25 0 100 0
25 – 61 0 100 ª 0 0 ª 10061 – 80 0 0 100
Reaction reagent: Mix 407 g of lithium acetate dihydrate,245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-propanol, add water to make 2000 mL, stir for more than 10minutes while passing Nitrogen, and use this solution as So-lution A. Separately, to 1957 mL of 1-methoxy-2-propanoladd 77 g of ninhydrin and 0.134 g of sodium borohydride,stir for more than 30 minutes while passing Nitrogen, anduse this solution as Solution B. Mix Solution A and SolutionB before use.
Flow rate of mobile phase: About 0.26 mL per minute.Flow rate of reaction reagent: About 0.3 mL per minute.
System suitability—System performance: When the procedure is run with
20 mL of the standard solution under the above operatingconditions, aspartic acid, threonine, serine, glutamic acid,proline, glycine, alanine, valine, cystine, methionine, isoleu-cine, leucine, tyrosine, phenylalanine, lysine, histidine andarginine are eluted in this order with the resolutions betweenthe peaks of threonine and serine, glycine and alanine, andisoleucine and leucine being not less than 1.5, 1.4 and 1.2,respectively.
System repeatability: When the test is repeated 3 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peakarea of aspartic acid, proline, valine and arginine are notmore than 2.0z, respectively.
Purity (1) Acetic acid—Weigh accurately about 15 mg ofOxytocin, dissolve in the internal standard solution to makeexactly 10 mL, and use this solution as the sample solution.Separately, weigh accurately about 1 g of acetic acid (100),add the internal standard solution to make exactly 100 mL.Pipet 2 mL of this solution, add the internal standardsolution to make exactly 200 mL, and use this solution as thestandard solution. Perform the test with 10 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and calculate the ratios, QT and QS, of the peakarea of acetic acid to that of the internal standard: theamount of acetic acid is not less than 6.0z and not morethan 10.0z.
Amount (z) of acetic acid (C2H4O2)
＝WS
WT×
QT
QS×
110
WS: Amount (mg) of acetic acid (100)WT: Amount (mg) of the sample
Internal standard solution—A solution of propionic acid inthe mobile phase (1 in 10,000).
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 210 nm).Column: A stainless steel column 4.6 mm in inside di-
ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about409C.
Mobile phase: To 0.7 mL of phosphoric acid add 900 mLof water, adjust the pH to 3.0 with 8 mol/L sodiumhydroxide TS, and add water to make 1000 mL. To 950 mLof this solution add 50 mL of methanol.
Flow rate: Adjust the ‰ow rate so that the retention timeof acetic acid is about 3 minutes.System suitability—
System performance: When the procedure is run with 10mL of the standard solution under the above operatingconditions, acetic acid and propionic acid are eluted in thisorder with the resolution between these peaks being not lessthan 14.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of acetic acid to that of the internal standard isnot more than 2.0z.
(2) Related substances—Dissolve 25 mg of Oxytocin in100 mL of the mobile phase A, and use this solution as thesample solution. Perform the test with 50 mL of the samplesolution as directed under the Liquid Chromatographyaccording to the following conditions, determine each peakarea by the automatic integration method, and calculate theamount of them by the area percentage method: the amountof each peak other than Oxytocin is not more than 1.5z,and the total of them is not more than 5.0z.Operating conditions—
Detector, column, column temperature, mobile phase,‰owing of mobile phase, and ‰ow rate: Proceed as directedin the operating conditions in the Assay.
Time span of measurement: About 2.5 times as long as theretention time of oxytocin.System suitability—
Test for required detectability: Measure exactly 1 mL ofthe sample solution, add the mobile phase A to make exactly100 mL, and use this solution as the solution for systemsuitability test. Pipet 1 mL of the solution for systemsuitability test, and add the mobile phase A to make exactly10 mL. Conˆrm that the peak area of oxytocin obtainedfrom 50 mL of this solution is equivalent to 5 to 15z of thatfrom 50 mL of the solution for system suitability test.
System performance: Dissolve an adequate amount ofoxytocin and vasopressin in the mobile phase A, so that eachmL contains about 0.1 mg each of them. When the proce-dure is run with 50 mL of this solution under the above oper-ating conditions, vasopressin and oxytocin are eluted in thisorder with the resolution between these peaks being not lessthan 14, and the symmetry factor of the peak of oxytocin isnot more than 1.5.
17301730 Supplement II, JPXIVO‹cial Monographs for Part I
System repeatability: When the test is repeated 6 timeswith 50 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of oxytocin is not more than 2.0z.
Water Not more than 5.0z (50 mg, coulometric titration).
Assay Weigh accurately an amount of Oxytocin, equiva-lent to about 13,000 Units, dissolve in the mobile phase A tomake exactly 100 mL, and use this solution as the samplesolution. Separately, dissolve oxytocin in 1 bottle of thePosterior Pituitary Reference Standard in the mobile phaseA to make a known concentration solution containing eachmL contains about 130 Units, and use this solution as thestandard solution. Perform the test with exactly 25 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the peak areas, AT and AS, ofoxytocin.
Units per mg of Oxytocin, calculated on the dehydrated andde-acetic acid basis
＝WS
WT×
AT
AS×100
WS: Units per mL of the standard solutionWT: Amount (mg) of the sample, calculated on the de-
hydrated and de-acetic acid basis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 220 nm).Column: A stainless steel column 4.6 mm in inside di-
ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about259C.
Mobile phase A: Dissolve 15.6 g of sodium dihydrogenphosphate dihydrate in 1000 mL of water.
Mobile phase B: A mixture of water and acetonitrile (1:1).Flowing of the mobile phase: Control the gradient by
mixing the mobile phases A and B as directed in thefollowing table.
Flow rate: About 1.0 mL per minute.System suitability—
System performance: Dissolve an adequate amount ofoxytocin and vasopressin in the mobile phase A, so that eachmL contains about 0.1 mg each of them. When the proce-dure is run with 25 mL of this solution under the above oper-ating conditions, vasopressin and oxytocin are eluted in thisorder with the resolution between these peaks being not lessthan 14, and the symmetry factor of the peak of oxytocin is
not more than 1.5.System repeatability: When the test is repeated 6 times
with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of oxytocin is not more than 1.0z.
Containers and storage Containers—Tight containers.Storage—At 2 to 89C.
Change to read:
Oxytocin Injectionオキシトシン注射液
Oxytocin Injection is an aqueous solution for injec-tion.
It contains not less than 90.0z and not more than110.0z of the labeled oxytocin Units.
Method of preparation Prepare as directed under Injec-tions, with Oxytocin.
Description Oxytocin Injection is a colorless, clear liquid.
pH 2.5 – 4.5
Bacterial endotoxins Less than 10 EU/oxytocin Unit.
Actual volume It meets the requirements of the Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay Measure exactly a portion of Oxytocin Injectionaccording to the labeled Units, dilute with the diluent so thateach mL contains about 1 Unit, and use this solution as thesample solution. Separately, dissolve oxytocin in 1 bottle ofPosterior Pituitary Reference Standard in the mobile phaseA to make exactly 20 mL. Pipet a suitable volume of this so-lution, dilute with the diluent to make a known concentra-tion solution so that each mL contains about 1 Unit, and usethis solution as the standard solution. Perform the test withexactly 100 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine the peakareas, AT and AS, of oxytocin.
Units per mL of Oxytocin Injection＝WS×AT
AS×
ba
WS: Units per mL of the standard solutiona: Volume (mL) of the sampleb: Total volume of the sample solution prepared by dilut-
17311731Supplement II, JPXIV O‹cial Monographs for Part I
ing with the diluentDiluent: Dissolve 5 g of chlorobutanol, 1.1 g of sodium
acetate trihydrate, 5 g of acetic acid (100) and 6mL of ethanol (99.5) in water to make 1000 mL.
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 220 nm).Column: A stainless steel column 4.6 mm in inside di-
ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about259C.
Mobile phase A: Dissolve 15.6 g of sodium dihydrogenphosphate dihydrate in 1000 mL of water.
Mobile phase B: A mixture of water and acetonitrile (1:1).Flowing of the mobile phase: Control the gradient by
mixing the mobile phases A and B as directed in the follow-ing table.
Flow rate: About 1.0 mL per minute.System suitability—
System performance: Dissolve an adequate amount ofoxytocin and vasopressin in the mobile phase A, so that eachmL contains about 0.02 mg each of them. When the proce-dure is run with 100 mL of this solution under the aboveoperating conditions, vasopressin and oxytocin are eluted inthis order with the resolution between these peaks being notless than 14, and the symmetry factor of the peak of oxyto-cin is not more than 1.5.
System repeatability: When the test is repeated 6 timeswith 100 mL of the standard solution under the above oper-ating conditions, the relative standard deviation of the peakarea of oxytocin is not more than 2.0z.
Containers and storage Containers—Hermetic containers.Storage—In a cold place, and avoid freezing.
Pirenoxineピレノキシン
Change the Purity (2) to read:
Purity(2) Related substances—Dissolve 10 mg of Pirenoxine in
50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution asthe standard solution. Perform the test with exactly 5 mLeach of the sample solution and the standard solution as
directed under the Liquid Chromatography according to thefollowing conditions. Determine each peak area of both so-lutions by the automatic integration method: the total areaof the peaks other than pirenoxine is not larger than the peakarea of pirenoxine from the standard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 230 nm).
Column: A stainless steel column 4 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about359C.
Mobile phase: Dissolve 1.39 g of tetra n-butylammoniumchloride and 4.5 g of disodium hydrogen phosphate 12-water in 1000 mL of water, and adjust the pH to 6.5 withphosphoric acid. To 700 mL of this solution add 200 mL ofacetonitrile and 30 mL of tetrahydrofuran, and mix.
Flow rate: Adjust the ‰ow rate so that the retention timeof pirenoxine is about 10 minutes.
Time span of measurement: About 3 times as long as theretention time of pirenoxine.System suitability—
Test for required detectability: To exactly 2 mL of thestandard solution add the mobile phase to make exactly 30mL. Conˆrm that the peak area of pirenoxine obtained from5 mL of this solution is equivalent to 5 to 8z of that ofpirenoxine obtained from 5 mL of the standard solution.
System performance: Dissolve 3 mg of Pirenoxine and16 mg of methyl parahydroxybenzoate in 100 mL of themobile phase. When the procedure is run with 5 mL of thissolution under the above operating conditions, pirenoxineand methyl parahydroxybenzoate are eluted in this orderwith the resolution between these peaks being not less than2.0.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of pirenoxine is not more than 1.0z.
17321732 Supplement II, JPXIVO‹cial Monographs for Part I
Pirenzepine Hydrochloride Hydrate contains notless than 98.5z and not more than 101.0z of pirenze-pine hydrochloride (C19H21N5O2.2HCl: 424.32), calcu-lated on the anhydrous basis.
Description Pirenzepine Hydrochloride Hydrate occurs asa white to pale yellow crystalline powder.
It is freely soluble in water and in formic acid, slightlysoluble in methanol, and very slightly soluble in ethanol(99.5).
The pH of a solution by dissolving 1 g of PirenzepineHydrochloride Hydrate in 10 mL of water is between 1.0 and2.0.
Melting point: about 2459C (with decomposition).It is gradually colored by light.
Identiˆcation (1) Determine the absorption spectrum ofa solution of Pirenzepine Hydrochloride Hydrate (1 in40,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofPirenzepine Hydrochloride Hydrate as directed in the potas-sium chloride disk method under the Infrared Spectrophoto-metry, and compare the spectrum with the ReferenceSpectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(3) A solution of Pirenzepine Hydrochloride Hydrate (1in 50) responds to the Qualitative Tests for chloride.
Purity (1) Clarity and color of solution—A solutionobtained by dissolving 1.0 g of Pirenzepine HydrochlorideHydrate in 10 mL of water is clear and not more color thanthat of the following control solution.
Control solution: To 1.2 mL of Matching ‰uid for color Fadd 8.8 mL of diluted hydrochloric acid (1 in 40).
(2) Heavy metals—Proceed with 2.0 g of PirenzepineHydrochloride Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of
Standard Lead Solution (not more than 10 ppm).(3) Related substances—Dissolve 0.3 g of Pirenzepine
Hydrochloride Hydrate in 10 mL of water. To 1 mL of thissolution add 5 mL of methanol and the mobile phase A tomake 10 mL, and use this solution as the sample solution.Pipet 1 mL of the sample solution, and add 5 mL ofmethanol and the mobile phase A to make exactly 10 mL.Pipet 1 mL of this solution, add 5 mL of methanol and themobile phase A to make exactly 10 mL, and use this solutionas the standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the area of the peak otherthan pirenzepine is not more than 3/10 times the peak areaof pirenzepine from the standard solution, and the total areaof the peaks other than pirenzepine is not more than 3/5times the peak area of pirenzepine from the standard solu-tion.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 283 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase A: Dissolve 2 g of sodium lauryl sulfate in900 mL of water, adjust the pH to 3.2 with acetic acid (100),and add water to make 1000 mL.
Mobile phase B: MethanolMobile phase C: AcetonitrileFlowing of the mobile phase: Control the gradient by
mixing the mobile phases A, B and C as directed in the fol-lowing table.
Time after injectionof sample (min)
Mobile phaseA (z)
Mobile phaseB (z)
Mobile phaseC (z)
0 – 15 55 ª 25 30 15 ª 4515 – 25 30 45
Flow rate: Adjust the ‰ow rate so that the retention timeof pirenzepine is about 8 minutes.
Time span of measurement: About 2 times as long as theretention time of pirenzepine after the solvent peak.System suitability—
Test for required detectability: Pipet 1 mL of the standardsolution, and add 5 mL of methanol and the mobile phase Ato make exactly 10 mL. Conˆrm that the peak area of piren-zepine obtained from 10 mL of this solution is equivalent to 7to 13z of that from 10 mL of the standard solution.
System performance: Dissolve 0.1 g of phenylpiperazinehydrochloride in 10 mL of methanol. Mix 1 mL of thissolution and 1 mL of the sample solution, and add 5 mL ofmethanol and the mobile phase A to make 10 mL. When theprocedure is run with 10 mL of this solution under the above
17331733Supplement II, JPXIV O‹cial Monographs for Part I
operating conditions, pirenzepine and phenylpiperazine areeluted in this order with the resolution between these peaksbeing not less than 5.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the aboveoperating conditions, the relative standard deviation of thepeak area of pirenzepine is not more than 2.0z.
Water Not less than 3.5z and not more than 5.0z (0.3 g,volumetric titration, direct titration).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately about 0.2 g of PirenzepineHydrochloride Hydrate, dissolve in 2 mL of formic acid,add 60 mL of acetic anhydride, and titrate with 0.1 mol/Lperchloric acid VS (potentiometric titration). Perform ablank determination in the same manner, and make anynecessary correction.
Each mL of 0.1 mol/L perchloric acid VS＝14.14 mg of C19H21N5O2.2HCl
Containers and storage Containers—Well-closed contain-ers.
Piroxicam contains not less than 98.5z and notmore than 101.0z of C15H13N3O4S, calculated on thedried basis.
Description Piroxicam occurs as a white to pale yellowcrystalline powder.
It is sparingly soluble in acetic anhydride, slightly solublein acetonitrile, in methanol and in ethanol (99.5), very slight-ly soluble in acetic acid (100), and practically insoluble inwater.
Melting point: about 2009C (with decomposition).
Identiˆcation (1) Dissolve 0.1 g of Piroxicam in a mix-ture of methanol and 0.5 mol/L hydrochloric acid TS(490:1) to make 200 mL. To 1 mL of this solution add themixture of methanol and 0.5 mol/L hydrochloric acid TS(490:1) to make 100 mL. Determine the absorption spectrumof this solution as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with the
Reference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofPiroxicam as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.If any diŠerence appears between the spectra, dissolve thesample with dichloromethane, evaporate the solvent, dry theresidue on a water bath, and perform the test.
Purity (1) Heavy metals—Proceed with 1.0 g of Piroxic-am according to Method 2, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 20 ppm).
(2) Related substances—Dissolve 75 mg of Piroxicam in50 mL of acetonitrile for liquid chromatography, and usethis solution as the sample solution. Pipet 1 mL of thesample solution, add acetonitrile for liquid chromatographyto make exactly 10 mL. Pipet 1 mL of this solution, addacetonitrile for liquid chromatography to make exactly 50mL, and use this solution as the standard solution. Performthe test with exactly 20 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine each peak area by the automatic integrationmethod: the area of the peak other than piroxicam is notlarger than the peak area of piroxicam with the standard so-lution, and the total area of the peaks other than piroxicamis not larger than 2 times the peak area of piroxicam with thestandard solution.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 230 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: A mixture of 0.05 mol/L potassium di-hydrogen phosphate TS, pH 3.0 and acetonitrile for liquidchromatography (3:2).
Flow rate: Adjust the ‰ow rate so that the retention timeof piroxicam is about 10 minutes.
Time span of measurement: About 5 times as long as theretention time of piroxicam after the solvent peak.System suitability—
Test for required detectability: To exactly 5 mL of thestandard solution add acetonitrile for liquid chro-matography to make exactly 20 mL. Conˆrm that the peakarea of piroxicam obtained with 20 mL of this solution isequivalent to 17.5 to 32.5z of that with 20 mL of thestandard solution.
System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry factor of the peak of piroxicam are not less than 6000 and
17341734 Supplement II, JPXIVO‹cial Monographs for Part I
not more than 1.5, respectively.System repeatability: When the test is repeated 6 times
with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of piroxicam is not more than 2.0z.
Loss on drying Not more than 0.5z (1 g, 1059C, 3 hours).
Residue on ignition Not more than 0.2z (1 g).
Assay Weigh accurately about 0.25 g of Piroxicam, dis-solve in 60 mL of a mixture of acetic anhydride and aceticacid (100) (1:1), and titrate with 0.1 mol/L perchloric acidVS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correc-tion.
Each mL of 0.1 mol/L perchloric acid VS＝33.14 mg of C15H13N3O4S
Containers and storage Containers—Tight containers.
Pyrazinamideピラジナミド
Change the Description to read:
Description Pyrazinamide occurs as white crystals orcrystalline powder.
It is sparingly soluble in water and in methanol, andslightly soluble in ethanol (99.5) and in acetic anhydride.
Change the Purity to read:
Purity(1) Heavy metals—Proceed with 1.0 g of Pyrazinamide
according to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).
(2) Related substances—Dissolve 0.10 g of Pyrazin-amide in 10 mL of methanol, and use this solution as thesample solution. Pipet 1 mL of the sample solution, addmethanol to make exactly 200 mL, and use this solution asthe standard solution. Perform the test with these solutionsas directed under the Thin-layer Chromatography. Spot 20mL each of the sample solution and the standard solution ona plate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance ofabout 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spot other than theprincipal spot obtained from the sample solution is not moreintense than the spot from the standard solution.
Change the Assay to read:
Assay Weigh accurately about 0.1 g of Pyrazinamide,previously dried, dissolve in 50 mL of acetic anhydride, andtitrate with 0.1 mol/L perchloric acid VS (potentiometrictitration). Perform a blank determination in the same man-
ner, and make any necessary correction.
Each mL of 0.1 mol/L perchloric acid VS＝12.31 mg of C5H5N3O
Pyridoxine Hydrochloride塩酸ピリドキシン
Change the origin/limits of content to read:
Pyridoxine Hydrochloride, when dried, contains notless than 98.0z and not more than 101.0z ofC8H11NO3.HCl.
Change the Description to read:
Description Pyridoxine Hydrochloride occurs as a white topale yellow, crystalline powder.
It is freely soluble in water, slightly soluble in ethanol(99.5), and practically insoluble in acetic anhydride and inacetic acid (100).
It is gradually aŠected by light.Melting point: about 2069C (with decomposition).
Change the Identiˆcation to read:
Identiˆcation(1) Determine the absorption spectrum of a solution of
Pyridoxine Hydrochloride in 0.1 mol/L hydrochloric acidTS (1 in 100,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum or the spectrum of a solution ofPyridoxine Hydrochloride Reference Standard prepared inthe same manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofPyridoxine Hydrochloride as directed in the potassiumchloride disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Pyridoxine Hydrochloride ReferenceStandard: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(3) A solution of Pyridoxine Hydrochloride (1 in 10)responds to the Qualitative Tests for chloride.
Add the following next to Identiˆcation:
pH The pH of a solution prepared by dissolving 1.0 g ofPyridoxine Hydrochloride in 50 mL of water is between 2.5and 3.5.
Add the following next to Purity (2):
(3) Related substances—Dissolve 1.0 g of PyridoxineHydrochloride in 10 mL of water, and use this solution asthe sample solution. Pipet 2.5 mL of the sample solution,and add water to make exactly 100 mL. Pipet 1 mL of thissolution, add water to make exactly 10 mL, and use this
17351735Supplement II, JPXIV O‹cial Monographs for Part I
solution as the standard solution. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 2 mL each of the sample solution and the standardsolution on a plate of silica gel for thin-layer chro-matography, and air-dry the plate. Develop the plate with amixture of acetone, tetrahydrofuran, hexane and ammoniasolution (28) (65:13:13:9) to a distance of about 10 cm, andair-dry the plate. Spray evenly a solution of sodiumcarbonate in diluted ethanol (99.5) (3 in 10) (1 in 20) on theplate. After air-drying, spray evenly a solution of 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine in ethanol(99.5) (1 in 1000) on the plate, and air-dry: the spot otherthan the principal spot obtained from the sample solution isnot more intense than the spot from the standard solution.
Pyridoxine Hydrochloride Injection塩酸ピリドキシン注射液
Change the origin/limits of content to read:
Pyridoxine Hydrochloride Injection is an aqueoussolution for injection.
It contains not less than 95.0z and not more than105.0z of the labeled amount of pyridoxinehydrochloride (C8H11NO3.HCl: 205.64).
Change the Identiˆcation to read:
Identiˆcation(1) To a volume of Pyridoxine Hydrochloride Injection,
equivalent to 50 mg of Pyridoxine Hydrochloride accordingto the labeled amount, add 0.1 mol/L hydrochloric acid TSto make 100 mL. To 2 mL of this solution add 0.1 mol/Lhydrochloric acid TS to make 100 mL. Determine theabsorption spectrum of this solution as directed under theUltraviolet-visible spectrophotometry: it exhibits a maxi-mum between 288 nm and 292 nm.
(2) To a volume of Pyridoxine Hydrochloride Injection,equivalent to 10 mg of Pyridoxine Hydrochloride accordingto the labeled amount, add water to make 10 mL, and usethis solution as the sample solution. Separately, dissolve 10mg of Pyridoxine Hydrochloride Reference Standard in 10mL of water, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 2 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography, and air-dry the plate. Developthe plate with a mixture of acetone, tetrahydrofuran, hexaneand ammonia solution (28) (65:13:13:9) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly a solutionof sodium carbonate in diluted ethanol (99.5) (3 in 10) (1 in20) on the plate. After air-drying, spray evenly a solution of2,6-dibromo-N-chloro-1,4-benzoquinone monoimine inethanol (99.5) (1 in 1000) on the plate: the spots obtainedfrom the sample solution and the standard solution are bluein color and have the same Rf value.
Add the following next to Identiˆcation:
Endotoxins Less than 3.0 EU/mg.
Delete the following Monograph:
Santonin Tablets
Add the following:
Serrapeptaseセラペプターゼ
[95077-02-4]
Serrapeptase is the enzyme preparation having pro-teolytic activity, produced by the growth of Serratiaspecies. Usually, it is diluted with Lactose.
It contains not less than 2000 serrapeptase Units andnot more than 2600 serrapeptase Units per mg.
It is hygroscopic.
Description Serrapeptase occurs as a grayish white to lightbrown powder, having a slight characteristic odor.
Identiˆcation Dissolve 0.4 g of Serrapeptase in 100 mL ofacetic acid-sodium acetate buŠer solution, pH 5.0, transferexactly 1 mL each of this solution into three tubes, and referto them as A, B and C. To tube A add exactly 1 mL of water,to tubes B and C add exactly 1 mL of 0.04 mol/L disodiumdihydrogen ethylenediamine tetraacetate TS, mix gently,and allow them to stand in a water bath at 4±19C for about1 hour. Then, to the tube B add exactly 2 mL of 0.04 mol/Lzinc chloride TS, to the tubes A and C add exactly 2 mL ofwater, mix gently, and allow them to stand in a water bath at4±19C for about 1 hour. Pipet 1 mL each of these solu-tions, add borate-hydrochloric acid buŠer solution, pH 9.0to the solutions A and B to make exactly 200 mL, to the so-lution C to make exactly 50 mL, and use these solutions asthe sample solutions. Proceed with these sample solutions asdirected in the Assay: the activities of the solutions A and Bare almost the same, and the activity of the solution C is notmore than 5z of that of the solution A.
Activity of solutions A, B or C＝AT
AS×
120×D×176
AS: Absorbance of the standard solutionAT: Absorbance of the sample solution20: Reaction time (minute)D: Dilution rate (200 for solution A and B, 50 for solution
C)176: Conversion factor (Total volume of enzyme reaction
solution/volume of ˆltrate taken×amount of tyrosine in 2mL of tyrosine standard solution)
Purity (1) Heavy metals—Put 1.0 g of Serrapeptase in a
17361736 Supplement II, JPXIVO‹cial Monographs for Part I
porcelain crucible, add 2 drops each of sulfuric acid andnitric acid, and incinerate by ignition. After cooling, to theresidue add 2 mL of hydrochloric acid, evaporate to drynesson a water bath, add 10 mL of a solution of hydroxylaminehydrochloride (3 in 100) and 2 mL of dilute acetic acid, andheat on a water bath for 5 minutes. After cooling, ˆlter ifnecessary, wash the ˆlter paper with 10 mL of water, put theˆltrate and washing in a Nessler tube, add water to make 50mL, and use this solution as the test solution. Prepare thecontrol solution as follows: Evaporate to dryness 2 dropseach of sulfuric acid and nitric acid on a sand bath, add 2mL of hydrochloric acid to the residue, evaporate to drynesson a water bath, add 2.0 mL of Standard Lead Solution, 10mL of a solution of hydroxylamine hydrochloride (3 in 100)and 2 mL of dilute acetic acid, and heat on a water bath for5 minutes. Proceed in the same manner as directed for thepreparation of the test solution, and add water to make 50mL (not more than 20 ppm).
(2) Arsenic—Prepare the test solution with 0.40 g ofSerrapeptase according to Method 3, excepting addition of 5mL of a solution of magnesium nitrate hexahydrate inethanol (95) (3 in 10) instead of a solution of magnesium ni-trate hexahydrate in ethanol (95) (1 in 50), evaporating todryness on a water bath, then incinerating with a small‰ame, and perform the test (not more than 5 ppm).
Loss on drying Not more than 7.0z (1 g, 1059C, 4 hours).
Residue on ignition Not more than 1.5z (1 g).
Assay (i) Sample solution: Dissolve exactly 0.100 g ofSerrapeptase in a solution of ammonium sulfate (1 in 20) tomake exactly 100 mL. Pipet 1 mL of this solution, addborate-hydrochloric acid buŠer solution, pH 9.0 to makeexactly 200 mL, and use this solution as the sample solution.
(ii) Tyrosine standard solution: Dissolve exactly 0.160 gof Tyrosine Reference Standard, previously dried at 1059Cfor 3 hours, in 0.2 mol/L hydrochloric acid TS to makeexactly 1000 mL. Pipet 10 mL of this solution, and add0.2 mol/L hydrochloric acid TS to make exactly 100 mL.Prepare before use.
(iii) Substrate solution: Previously determine the loss ondrying (609C, reduced pressure not exceeding 0.67 kPa, 3hours) of milk casein, previously dried. To exactly 1.20 g ofthe milk casein, calculated based on the loss on drying, add160 mL of a solution of sodium borate (19 in 1000), and heatin a water bath to dissolve. After cooling, adjust the pH toexactly 9.0 with 1 mol/L hydrochloric acid TS, and addborate-hydrochloric acid buŠer solution, pH 9.0 to make ex-actly 200 mL. Use after warming to 37±0.59C. Prepare be-fore use.
(iv) Precipitation reagent: Trichloroacetic acid TS forserrapeptase. Use after warming to 37±0.59C.
(v) Procedure: Pipet 1 mL of the sample solution, put ina glass-stoppered tube (15×130 mm), allow to stand at 37±0.59C for 5 minutes, add exactly 5 mL of the substratesolution, and mix well immediately. Allow to stand at 37±0.59C for exactly 20 minutes, add exactly 5 mL oftrichloroacetic acid TS for serrapeptase, mix, allow to stand
at 37±0.59C for 30 minutes, and ˆlter through a dried ˆlterpaper. Pipet 2 mL of the ˆltrate, add exactly 5 mL of asolution of anhydrous sodium carbonate (3 in 50), mix, addexactly 1 mL of diluted Folin's TS (1 in 3), mix well, andallow to stand at 37±0.59C for 30 minutes. Determine theabsorbance of this solution at 660 nm, A1, as directed underthe Ultraviolet-visible Spectrophotometry using water as theblank. Separately, pipet 1 mL of the sample solution, addexactly 5 mL of trichloroacetic acid TS for serrapeptase,mix, add exactly 5 mL of the substrate solution, allow tostand at 37±0.59C for 30 minutes, and proceed in the samemanner as directed above to determine the absorbance A2.Separately, pipet 2 mL of the tyrosine standard solution,add exactly 5 mL of a solution of anhydrous sodiumcarbonate (3 in 50), mix, add exactly 1 mL of diluted Folin'sTS (1 in 3), mix well, and proceed in the same manner asdirected above to determine the absorbance A3. Separately,pipet 2 mL of 0.2 mol/L hydrochloric acid TS, and proceedin the same manner as directed above to determine theabsorbance A4.
Serrapeptase Unit per mg of Serrapeptase
＝A1－A2
A3－A4×
120×200×176
20: Reaction time (minute)200: Dilution rate176: Conversion factor (Total volume of enzyme reaction
solution/volume of ˆltrate taken×amount of tyro-sine in 2 mL of tyrosine standard solution)
One serrapeptase Unit corresponds to the amount ofserrapeptase which produces 5 mg of tyrosine per minutefrom 5 mL of the substrate solution under the above condi-tions.
Containers and storage Containers—Tight containers.
Sodium Aurothiomalate金チオリンゴ酸ナトリウム
Change the origin/limits of content to read:
Sodium Aurothiomalate contains not less than49.0z and not more than 52.5z of gold (Au: 196.97),calculated on the anhydrous basis and corrected by theamount of ethanol.
Change the Description to read:
Description Sodium Aurothiomalate occurs as white tolight yellow, powder or granules.
It is very soluble in water, and practically insoluble inethanol (99.5).
It is hygroscopic.It changes in color by light to greenish pale yellow.
Change the Identiˆcation (3) to read:Identiˆcation
17371737Supplement II, JPXIV O‹cial Monographs for Part I
(3) Place 2 mL of a solution of Sodium Aurothiomalate(1 in 10) in a porcelain crucible, add 1 mL of ammonia TSand 1 mL of hydrogen peroxide (30), evaporate to dryness,and ignite. Add 20 mL of water to the residue, and ˆlter: theresidue on the ˆlter paper occurs as a yellow or dark yellow,powder or granules.
Add the following next to Identiˆcation (3):
Identiˆcation(4) The ˆltrate obtained in (3) responds to the Qualita-
tive Tests for sodium salt.(5) The ˆltrate obtained in (3) responds to the Qualita-
tive Tests for sulfate.
Add the following next to Purity (3):
Purity(4) Ethanol—Weigh accurately about 0.2 g of Sodium
Aurothiomalate, add exactly 3 mL of the internal standardsolution and 2 mL of water to dissolve, and use this solutionas the sample solution. Separately, pipet 3 mL of ethanol(99.5), and add water to make exactly 1000 mL. Pipet 2 mLof this solution, add exactly 3 mL of the internal standardsolution, and use this solution as the standard solution.Perform the test with 2 mL each of the sample solution andthe standard solution as directed under the Gas Chro-matography according to the following conditions, anddetermine the ratios of the peak area of ethanol to that ofthe internal standard, QT and QS: the amount of ethanol isnot more than 3.0z.
Amount (mg) of ethanol＝QT
QS×6×0.793
0.793: Density (g/mL) of ethanol (99.5) at 209C
Internal standard solution—A solution of 2-propanol (1 in500).Operating conditions—
Detector: Hydrogen ‰ame-ionization detector.Column: A column 3 mm in inside diameter and 3 m in
length, packed with porous styrene-divinylbenzenecopolymer for gas chromatography (particle diameter: 150 –180 mm) (average pore size: 0.0085 mm; 300 – 400 m2/g).
Column temperature: A constant temperature of about1809C.
Carrier gas: NitrogenFlow rate: Adjust the ‰ow rate so that the retention time
of the internal standard is about 7 minutes.System suitability—
System performance: When the procedure is run with 2 mLof the standard solution under the above operating condi-tions, ethanol and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 4.
System repeatability: When the test is repeated 6 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of ethanol to that of the internal standard is
not more than 2.0z.
Add the following next to Purity:
Water Not more than 5.0z (0.1 g, coulometric titration).Use a water vaporizer (heating temperature: 1059C; heatingtime: 30 minutes).
Delete the Loss on drying:
Change the Assay to read:
Assay Weigh accurately about 25 mg of SodiumAurothiomalate, and dissolve in 2 mL of aqua regia by heat-ing. After cooling, add water to make exactly 100 mL. Pipet2 mL of the solution, add water to make exactly 25 mL, anduse this solution as the sample solution. Separately, pipet 5mL, 10 mL and 15 mL of Standard Gold Solution for atom-ic absorption spectrophotometry, add water to make exactly25 mL, and use these solutions as the standard solutions (1),(2) and (3), respectively. Perform the test with the sample so-lution and the standard solutions (1), (2) and (3) as directedunder the Atomic Absorption Spectrophotometry under thefollowing conditions. Determine the amount of gold in thesample solution using the calibration curve obtained fromthe absorbances of the standard solutions.
(4) Styrene—To 10.0 g of Sodium Polystyrene Sulfonateadd 10 mL of acetone, shake for 30 minutes, centrifuge, anduse the supernatant liquid as the sample solution. Separate-ly, dissolve 10 mg of styrene in acetone to make exactly 100mL. Pipet 1 mL of this solution, add acetone to makeexactly 100 mL, and use this solution as the standardsolution. Perform the test with 20 mL each of the sample so-lution and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine peak areas, AT and AS, of styrene in each solu-tion: AT is not larger than AS.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of water and acetonitrile (1:1).Flow rate: Adjust the ‰ow rate so that the retention time
17381738 Supplement II, JPXIVO‹cial Monographs for Part I
of styrene is about 8 minutes.System suitability—
System performance: Dissolve 20 mg each of styrene andbutyl parahydroxybenzoate in 100 mL of acetone. To 5 mLof this solution add acetone to make 100 mL. When theprocedure is run with 20 mL of this solution under the aboveoperating conditions, butyl parahydroxybenzoate and sty-rene are eluted in this order with the resolution betweenthese peaks being not less than 5.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of styrene is not more than 2.0z.
Sodium Thiosulfateチオ硫酸ナトリウム
Change the origin/limits of content to read:
Sodium Thiosulfate, when dried, contains not lessthan 99.0z and not more than 101.0z of sodiumthiosulfate (Na2S2O3: 158.11).
Change the Description to read:
Description Sodium Thiosulfate occurs as colorless, crys-tals or crystalline powder. It is odorless.
It is very soluble in water, and practically insoluble inethanol (99.5).
It eŒoresces in dry air, and is deliquescent in moist air.
Change the Identiˆcation to read:
Identiˆcation(1) A solution of Sodium Thiosulfate (1 in 10) responds
to the Qualitative Tests for thiosulfate.(2) A solution of Sodium Thiosulfate (1 in 10) responds
to the Qualitative Tests for sodium salt.
Change the Loss on drying to read:
Loss on drying 32.0 – 37.0z (1 g, in vacuum, 40 – 459C,16 hours).
Change the Assay to read:
Assay Weigh accurately about 0.4 g of Sodium Thiosul-fate, previously dried, dissolve in 30 mL of water, and titratewith 0.05 mol/L iodine VS (indicator: 1 mL of starch TS).
Each mL of 0.05 mol/L iodine VS＝15.81 mg of Na2S2O3
Tegafurテガフール
Add the following next to Identiˆcation:
pH Dissolve 0.5 g of Tegafur in 50 mL of water: the pH ofthis solution is between 4.2 and 5.2.
Testosterone Propionateプロピオン酸テストステロン
Change the Description to read:
Description Testosterone Propionate occurs as white topale yellow crystals or crystalline powder.
It is freely soluble in methanol and in ethanol (95), andpractically insoluble in water.
Change the Identiˆcation to read:
Identiˆcation(1) Determine the absorption spectrum of a solution of
Testosterone Propionate in ethanol (95) (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Testosterone PropionateReference Standard prepared in the same manner as thesample solution: both spectra exhibit similar intensities ofabsorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofTestosterone Propionate, previously dried, as directed in thepotassium bromide disk method under the InfraredSpectrophotometry, and compare the spectrum with theReference Spectrum or the spectrum of TestosteronePropionate Reference Standard: both spectra exhibit similarintensities of absorption at the same wave numbers.
Purity Other steroids—Dissolve 40 mg of TestosteronePropionate in 2 mL of ethanol (95), and use this solution asthe sample solution. Pipet 1 mL of this solution, add ethanol(95) to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 10 mLeach of the sample solution and the standard solution on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture of chlo-roform and diethylamine (19:1) to a distance of about 15cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 254 nm): the spot other than the principal
17391739Supplement II, JPXIV O‹cial Monographs for Part I
spot from the sample solution is not more intense than thespot from the standard solution.
Change the Assay to read:
Assay Weigh accurately each about 10 mg of TestosteronePropionate and Testosterone Propionate ReferenceStandard, previously dried, and dissolve in methanol tomake exactly 100 mL. To exactly 5 mL of these solutionsadd exactly 5 mL of the internal standard solution andmethanol to make 20 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith 5 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of testosteronepropionate to that of the internal standard.
Amount (mg) of C22H32O3
＝WS×QT
QS
WS: Amount (mg) of Testosterone Propionate ReferenceStandard
Internal standard solution—A solution of Progesterone inmethanol (9 in 100,000).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 241 nm).
Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).
Column temperature: A constant temperature of about359C.
Mobile phase: A mixture of acetonitrile and water (7:3).Flow rate: Adjust the ‰ow rate so that the retention time
of testosterone propionate is about 10 minutes.System suitability—
System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, the internal standard and testosterone propionate areeluted in this order with the resolution between these peaksbeing not less than 9.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of testosterone propionate to that of theinternal standard is not more than 1.0z.
Testosterone Propionate Injectionプロピオン酸テストステロン注射液
Change the origin/limits of content to read:
Testosterone Propionate Injection is an oily solutionfor injection.
It contains not less than 92.5z and not more than107.5z of the labeled amount of testosteronepropionate (C22H32O3: 344.49).
Change the Identiˆcation to read:
Identiˆcation Dissolve the residue obtained as directed inthe procedure in the Assay in exactly 20 mL of methanol,and use this solution as the sample solution. Separately,dissolve 1 mg of Testosterone Propionate ReferenceStandard in 10 mL of methanol, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 10 mLeach of the sample solution and the standard solution on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture ofchloroform and diethylamine (19:1) to a distance of about15 cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 254 nm): the Rf values of the principalspot with the sample solution and of the spot with the stan-dard solution are not diŠerent each other.
Add the following next to Identiˆcation:
Actual volume It meets the requirements of Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Change the Assay to read:
Assay(i) Chromatographic tube A glass tube about 1 cm in
inside diameter and about 18 cm in length, with a glass ˆlter(G3) at the lower end.
(ii) Chromatographic column To about 2 g of silica gelfor liquid chromatography add 5 mL of dichloromethane,and mix gently. Transfer and wash into the chromatographictube with the aid of dichloromethane, allow to elute thedichloromethane through the column, and put a ˆlter paperon the upper end of the silica gel.
(iii) Standard solution Weigh accurately about 10 mgof Testosterone Propionate Reference Standard, previouslydried at 1059C for 4 hours, and dissolve in methanol tomake exactly 100 mL. Pipet 5 mL of this solution, addexactly 5 mL of the internal standard solution and methanolto make exactly 20 mL.
(iv) Sample stock solution To exactly a volume ofTestosterone Propionate Injection, equivalent to about20 mg of testosterone propionate (C22H32O3), adddichloromethane to make exactly 20 mL.
(v) Procedure Transfer exactly 2 mL of the samplestock solution into the chromatographic column, andelute to the upper surface of the silica gel. Wash the innersurface of the chromatographic tube with 15 mL of
17401740 Supplement II, JPXIVO‹cial Monographs for Part I
dichloromethane, elute to the upper surface of the silica gel,and discard the eŒuent. Elute 15 mL of a mixture ofdichloromethane and methanol (39:1), discard the ˆrst5 mL of the eŒuent, and collect the subsequent eŒuent.Wash the lower part of the column with a few amount ofdichloromethane, combine the washings and the eŒuent,and evaporate the solvent under reduced pressure. Dissolvethe residue so obtained with methanol to make exactly 20mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution and methanol to make exactly 20mL, and use this solution as the sample solution. Performthe test with 5 mL each of the sample solution and the stan-dard solution as directed in the Assay under TestosteronePropionate.
Amount (mg) of testosterone propionate (C22H32O3)
＝WS×QT
QS×2
WS: Amount (mg) of Testosterone Propionate ReferenceStandard
Thiamine Hydrochloride塩酸チアミン
Change the Purity (5) to read:
Purity(5) Related substances—Dissolve 0.10 g of Thiamine
Hydrochloride in 100 mL of the mobile phase, and use thissolution as the sample solution. Pipet 1 mL of the samplesolution, add the mobile phase to make exactly 100 mL, anduse this solution, as the standard solution. Perform the testwith exactly 10 mL each of the sample solution and the thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine the area of each peak by the automatic integrationmethod: the total area of the peaks other than thiamine isnot larger than the peak area of thiamine from the standardsolution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 3 times as long as theretention time of thiamine.System suitability—
Test for required detectability: To exactly 5 mL of thestandard solution add water to make exactly 50 mL. Con-ˆrm that the peak area of thiamine obtained from 10 mL ofthis solution is equivalent to 7 to 13z of that of thiamineobtained from 10 mL of the standard solution.
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak
area of thiamine is not more than 1.0z.
Change the Assay to read:
Assay Weigh accurately about 0.1 g each of ThiamineHydrochloride and Thiamine Hydrochloride ReferenceStandard (previously determine the water content), anddissolve them in the mobile phase to make exactly 50 mL. To10 mL each of the solutions, accurately measured, addexactly 5 mL each of the internal standard solution, add themobile phase to make 50 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak area of thiamine to that of the inter-nal standard.
Amount (mg) of C12H17ClN4OS.HCl
＝WS×QT
QS
WS: Amount (mg) of Thiamine Hydrochloride ReferenceStandard, calculated on the anhydrous basis
Internal standard solution—A solution methyl benzoate inmethanol (1 in 50).Operating conditions—Detector: An ultraviolet absorption photometer(wavelength: 254 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: Dissolve 1.1 g of sodium 1-octanesulfonatein 1000 mL of diluted acetic acid (100) (1 in 100). To 600 mLof this solution add 400 mL of a mixture of methanol andacetonitrile (3:2).
Flow rate: Adjust the ‰ow rate so that the retention timeof thiamine is about 12 minutes.System suitability—
System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, thiamine and the internal standard are eluted inthis order with the resolution between these peaks being notless than 6.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of thiamine to that of the internal standard isnot more than 1.0z.
17411741Supplement II, JPXIV O‹cial Monographs for Part I
Thiamylal Sodiumチアミラールナトリウム
Change the origin/limits of content to read:
Thiamylal Sodium contains not less than 97.5z andnot more than 101.0z of C12H17N2NaO2S, calculatedon the dried basis.
Change the Description to read:
Description Thiamylal Sodium occurs as light yellow crys-tals or powder.
It is very soluble in water, and freely soluble in ethanol(95).
The pH of a solution of Thiamylal Sodium (1 in 10) isbetween 10.0 and 11.0.
It is hygroscopic.It is gradually decomposed by light.Its solution in ethanol (95) (1 in 10) shows no optical
rotation.
Change the Identiˆcation to read:
Identiˆcation(1) Determine the absorption spectrum of a solution of
Thiamylal Sodium in ethanol (95) (7 in 1,000,000) as direct-ed under the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewavelengths.
(2) Determine the infrared absorption spectrum ofThiamylal Sodium, previously dried, as directed in thepotassium bromide disk method under the InfraredSpectrophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wave numbers.
(3) A solution of Thiamylal Sodium (1 in 10) responds tothe Qualitative Tests for sodium salt.
Change the Purity (3) to read:
Purity(3) Related substances—Dissolve 0.10 g of Thiamylal
Sodium in 10 mL of ethanol (95), and use this solution as thesample solution. Pipet 1 mL and 3 mL of the sample solu-tion, add ethanol (95) to make exactly 200 mL, and use thesesolutions as the standard solution (1) and the standard solu-tion (2). Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 10 mL each ofthe sample solution, the standard solution (1) and thestandard solution (2) on a plate of silica gel for thin-layerchromatography, develop with a mixture of toluene,methanol and ethyl acetate (40:7:3) to a distance of about12 cm, and air-dry the plate. Allow the plate to stand iniodine vapor for a night: the spot appeared around Rf 0.1obtained with the sample solution is not more intense than
the spot with the standard solution (2), and the spot otherthan the principal spot, the spot at origin and the spot men-tioned above obtained with the sample solution is not moreintense than the spot with the standard solution (1).
Change the Assay to read:
Assay Weigh accurately about 0.25 g of Thiamylal Sodi-um, dissolve in 50 mL of methanol and 5 mL of dilutehydrochloric acid, and add methanol to make exactly 100mL. Pipet 10 mL of this solution, and add methanol tomake exactly 100 mL. Pipet 5 mL of this solution, addexactly 10 mL of the internal standard solution and themobile phase to make 200 mL, and use this solution as thesample solution. Separately, weigh accurately about 23 mgof Thiamylal Reference Standard, previously dried at 1059Cfor 1 hour, dissolve in 50 mL of methanol and 0.5 mL ofdilute hydrochloric acid, and add methanol to make exactly100 mL. Pipet 5 mL of this solution, add exactly 10 mL ofthe internal standard solution and the mobile phase to make200 mL, and use this solution as the standard solution.Perform the test with 20 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area ofthiamylal to that of the internal standard.
Amount (mg) of C12H17N2NaO2S＝WS×QT
QS×10×1.0864
WS: Amount (mg) of Thiamylal Reference Standard
Internal standard solution—A solution of phenyl benzoatein methanol (3 in 500).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 289 nm).
Column: A stainless steel column about 4 mm in insidediameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of methanol and 0.05 mol/Lacetic acid-sodium acetate buŠer solution, pH 4.6 (13:7).
Flow rate: Adjust the ‰ow rate so that the retention timeof thiamylal is about 6 minutes.System suitability—
System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, thiamylal and the internal standard are eluted inthis order with the resolution between these peaks being notless than 12.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of thiamylal to that of the internal standard isnot more than 1.0z.
17421742 Supplement II, JPXIVO‹cial Monographs for Part I
Change to read:
Thiamylal Sodium for Injection注射用チアミラールナトリウム
Thiamylal Sodium for Injection is a preparation forinjection which is dissolved before use.
It contains not less than 93.0z and not more than107.0z of the labeled amount of thiamylal sodium(C12H17N2NaO2S: 276.33).
Method of preparation Prepare as directed under Injec-tions, with 100 parts of Thiamylal Sodium and 7 parts ofDried Sodium Carbonate in mass.
Description Thiamylal Sodium for Injection occurs as lightyellow crystals, powder or masses.
It is hygroscopic.It is gradually decomposed by light.
Identiˆcation (1) To 1.0 g of Thiamylal Sodium forInjection add 20 mL of ethanol (95), shake vigorously, andˆlter. Dissolve the precipitate so obtained in 1 mL of water,and add 1 mL of barium chloride TS: a white precipitate isproduced. Centrifuge this solution, take oŠ the supernatantliquid, and to the precipitate add dilute hydrochloric aciddropwise: the precipitate dissolves with eŠervescence.
(2) To 50 mg of Thiamylal Sodium for Injection add100 mL of ethanol (95), shake vigorously, and ˆlter. To 3mL of the ˆltrate add ethanol (95) to make 200 mL.Determine the absorption spectrum of this solution as direct-ed under the Ultraviolet-visible Spectrophotometry: itexhibits maxima between 236 nm and 240 nm, and between287 nm and 291 nm.
pH The pH of a solution obtained by dissolving 1.0 g ofThiamylal Sodium for Injection in 40 mL of water is be-tween 10.5 and 11.5.
Purity Related substances—To 0.10 g of Thiamylal Sodi-um for Injection add 10 mL of ethanol (95), shake vigorous-ly, ˆlter, and use the ˆltrate as the sample solution. Proceedas diected in the Purity (3) under Thiamylal Sodium.
Bacterial endotoxins Less than 1.0 EU/mg.
Mass variation It meets the requirements of the MassVariation Test.
Foreign insoluble matter Perform the test according toMethod 2: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay Open carefully 10 containers of Thiamylal Sodiumfor Injection, dissolve the contents with water, wash out theinside of each container with water, combine them, and addwater to make exactly V mL so that each mL contains about5 mg of thiamylal sodium (C12H17N2NaO2S). Pipet 5 mL ofthis solution, and add 0.5 mL of dilute hydrochloric acidand methanol to make exactly 100 mL. Pipet 5 mL of thissolution, add exactly 10 mL of the internal standard solutionand the mobile phase to make 200 mL, and use this solutionas the sample solution. Proceed the test with the sample so-lution as directed in the Assay under Thiamylal Sodium.
Amount (mg) of thiamylal sodium (C12H17N2NaO2S) in 1container
＝WS×QT
QS×
V50×1.0864
WS: Amount (mg) of Thiamylal Reference Standard
Internal standard solution—A solution of phenyl benzoatein methanol (3 in 500).
Containers and storage Containers—Hermetic containers.Storage—Light-resistant.
Add the following:
Tiaramide Hydrochloride Tablets塩酸チアラミド錠
Tiaramide Hydrochloride Tablets contain not lessthan 95.0z and not more than 105.0z of the labeledamount of tiaramide (C15H18ClN3O3S: 355.84).
Method of preparation Prepare as directed under Tablets,with Tiaramide Hydrochloride.
Identiˆcation (1) Determine the absorption spectrum ofthe sample solution obtained in the Assay as directed underthe Ultraviolet-visible Spectrophotometry: it exhibitsmaxima between 285 nm and 289 nm, and between 292 nmand 296 nm.
(2) To a quantity of powdered Tiaramide HydrochlorideTablets, equivalent to 0.1 g of tiaramide according to thelabeled amount, add 10 mL of diluted ethanol (99.5) (7 in10), shake well, ˆlter, and use the ˆltrate as the samplesolution. Separately, dissolve 0.11 g of tiaramide hydrochlo-ride for assay in 10 mL of diluted ethanol (99.5) (7 in 10),and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 20 mL each of the sample solutionand the standard solution on a plate of silica gel for thin-lay-er chromatography, develop with a mixture of 1-butamol,water and acetic acid (100) (4:2:1) to a distance of about 10cm, and dry the plate at 1009C for 30 minutes. Spray evenlyDragendorŠ's TS for spraying followed by diluted nitric acid(1 in 50) on the plate: the principal spot obtained with thesample solution and the spot with the standard solution areyellow-red in color and have the same Rf value.
17431743Supplement II, JPXIV O‹cial Monographs for Part I
Content uniformity Perform the test according to thefollowing method: it meets the requirements of the Contentuniformity Test.
To 1 tablet of Tiaramide Hydrochloride Tablets add avolume of 0.1 mol/L hydrochloric acid TS, equivalent to3/5 volume of V mL which makes a solution so that each mLcontains about 1 mg of tiaramide (C15H18ClN3O3S) accord-ing to the labeled amount, shake for 60 minutes, then add0.1 mol/L hydrochloric acid TS to make exactly V mL, andˆlter. Discard the ˆrst 20 mL of the ˆltrate, pipet 5 mL ofthe subsequent ˆltrate, add water to make exactly 100 mL,and use this solution as the sample solution. Separately,weigh accurately about 55 mg of tiaramide hydrochloridefor assay, previously dried at 1059C for 3 hours, and dis-solve in 0.1 mol/L hydrochloric acid TS to make exactly 50mL. Pipet 5 mL of this solution, add water to make exactly100 mL, and use this solution as the standard solution.Determine the absorbances, AT and AS, of the sample solu-tion and the standard solution at 294 nm as directed underthe Ultraviolet-visible Spectrophotometry.
Amount (mg) of tiaramide (C15H18ClN3O3S)
＝WS×AT
AS×
V50×0.907
WS: Amount (mg) of tiaramide hydrochloride for assay
Dissolution Perform the test with 1 tablet of TiaramideHydrochloride Tablets at 50 revolutions per minute accord-ing to Method 2 under the Dissolution Test, using 900 mL ofwater as the dissolution medium. Withdraw 20 mL or moreof the dissolution medium 15 minutes after starting the testfor a 50-mg tablet or 30 minutes after starting the test for a100-mg tablet, and ˆlter through a membrane ˆlter with poresize of not more than 0.5 mm. Discard the ˆrst 10 mL of theˆltrate, pipet V mL of the subsequent ˆltrate, add water tomake exactly V? mL so that each mL contains about 56 mg oftiaramide (C15H18ClN3O3S) according to the labeled amount,and use this solution as the sample solution. Separately,weigh accurately about 15 mg of tiaramide hydrochloridefor assay, previously dried at 1059C for 3 hours, and dis-solve in water to make exactly 50 mL. Pipet 5 mL of thissolution, add water to make exactly 25 mL, and use this so-lution as the standard solution. Determine the absorbances,AT and AS, at 294 nm of the sample solution and the stan-dard solution as directed under the Ultraviolet-visibleSpectrophotometry: the dissolution rates for a 50-mg tabletin 15 minutes and a 100-mg tablet in 30 minutes are not lessthan 80z, respectively.
Dissolution rate (z) with respect to the labeled amount oftiaramide (C15H18ClN3O3S)
＝WS×AT
AS×
V?V×
1C×360×0.907
WS: Amount (mg) of tiaramide hydrochloride for assayC: Labeled amount (mg) of tiaramide (C15H18ClN3O3S) in
1 tablet
Assay Weigh accurately the mass of more than 20Tiaramide Hydrochloride Tablets, and powder. Weigh ac-
curately an amount of the powder, equivalent to about 0.1 gof tiaramide (C15H18ClN3O3S), add 60 mL of 0.1 mol/Lhydrochloric acid TS, shake for 30 minutes, add 0.1 mol/Lhydrochloric acid TS to make exactly 100 mL, and ˆlter.Discard the ˆrst 20 mL of the ˆltrate, pipet 5 mL of the sub-sequent ˆltrate, add water to make exactly 100 mL, and usethis solution as the sample solution. Separately, weigh ac-curately about 0.11 g of tiaramide hydrochloride for assay,previously dried at 1059C for 3 hours, and dissolve in 0.1mol/L hydrochloric acid TS to make exactly 100 mL. Pipet5 mL of this solution, add water to make exactly 100 mL,and use this solution as the standard solution. Determine theabsorbances, AT and AS, of the sample solution and thestandard solution at 294 nm as directed under the Ultrav-iolet-visible Spectrophotometry.
Amount (mg) of tiaramide (C15H18ClN3O3S)
＝WS×AT
AS×0.907
WS: Amount (mg) of tiaramide hydrochloride for assay
Containers and storage Containers—Tight containers.
Tinidazoleチニダゾール
Change the origin/limits of content to read:
Tinidazole, when dried, contains not less than98.5z and not more than 101.0z of C8H13N3O4S.
Change the Description to read:
Description Tinidazole occurs as a light yellow, crystallinepowder.
It is soluble in acetic anhydride and in acetone, sparinglysoluble in methanol, slightly soluble in ethanol (99.5), andvery slightly soluble in water.
Change the Identiˆcation to read:
Identiˆcation(1) Determine the absorption spectrum of a solution of
Tinidazole in methanol (1 in 50,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofTinidazole as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.
17441744 Supplement II, JPXIVO‹cial Monographs for Part I
Tizanidine Hydrochloride, when dried, containsnot less than 99.0z and not more than 101.0z ofC9H8ClN5S.HCl.
Description Tizanidine Hydrochloride occurs as a white tolight yellowish white crystalline powder.
It is soluble in water, slightly soluble in ethanol (99.5), andpractically insoluble in acetic anhydride and in acetic acid(100).
Melting point: about 2909C (with decomposition).
Identiˆcation (1) Determine the absorption spectrum ofa solution of Tizanidine Hydrochloride in diluted 1 mol/Lammonia TS (1 in 10) (1 in 125,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.
(2) Determine the infrared absorption spectrum ofTizanidine Hydrochloride as directed in the potassium chlo-ride disk method under the Infrared Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.
(3) A solution of Tizanidine Hydrochloride (1 in 50)responds to the Qualitative Tests for chloride.
Purity (1) Heavy metals—Proceed with 1.0 g of Tizani-dine Hydrochloride according to Method 3, and perform thetest. Prepare the control solution with 2.0 mL of StandardLead Solution (not more than 20 ppm).
(2) Related substances—Dissolve 60 mg of TizanidineHydrochloride in 10 mL of a mixture of water and acetoni-trile (17:3), and use this solution as the sample solution.Pipet 1 mL of the sample solution, add the mixture of waterand acetonitrile (17:3) to make exactly 200 mL, and use thissolution as the standard solution. Perform the test with ex-actly 10 mL of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine each peak area bythe automatic integration method: the area of the peak otherthan tizanidine is not larger than 1/5 times the peak area oftizanidine with the standard solution.Operating conditions—
Detector: An ultraviolet absorption photometer
(wavelength: 230 nm for about 3 minutes after sample injec-tion and 318 nm subsequently).
Column: A stainless steel column 4.6 mm in insidediameter and 12.5 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase A: A mixture of water and formic acid(200:1), adjusted to pH 8.5 with ammonia water (28).
Mobile phase B: A mixture of acetonitrile and the mobilephase A (4:1).
Flowing of the mobile phase: Control the gradient bymixing the mobile phases A and B as directed in the follow-ing table.
Flow rate: Adjust the ‰ow rate so that the retention timeof tizanidine is about 7 minute.
Time span of measurement: About 4 times as long as theretention time of tizanidine after the solvent peak.System suitability—
Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the mixture of water andacetonitrile (17:3) to make exactly 10 mL. Conˆrm that thepeak area of tizanidine obtained with 10 mL of this solutionis equivalent to 14 to 26z of that with 10 mL of the standardsolution.
System performance: Dissolve 2 mg each of TizanidineHydrochloride and p-toluenesulfonic acid monohydrate in100 mL of the mixture of water and acetonitrile (17:3).When the procedure is run with 10 mL of this solution underthe above operating conditions, p-toluenesulfonic acid andtizanidine are eluted in this order with the resolutionbetween these peaks being not less than 10.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tizanidine is not more than 2.0z.
Loss on drying Not more than 0.2z (1 g, 1059C, 3 hours).
Residue on ignition Not more than 0.1z (1 g).
Assay Weigh accurately about 0.2 g of TizanidineHydrochloride, previously dried, dissolve in 60 mL of amixture of acetic anhydride and acetic acid (100) (7:3) withthe aid of warming. After cooling, titrate with 0.1 mol/Lperchloric acid VS (potentiometric titration). Perform ablank determination in the same manner, and make anynecessary correction.
17451745Supplement II, JPXIV O‹cial Monographs for Part I
Each mL of 0.1 mol/L perchloric acid VS＝29.02 mg of C9H8ClN5S.HCl
Containers and storage Containers—Well-closed contain-ers.
Tranexamic Acidトラネキサム酸
Change the origin/limits of content to read:
Tranexamic Acid, when dried, contains not less than98.0z and not more than 101.0z of C8H15NO2.
Change the Description to read:
Description Tranexamic Acid occurs as white crystals orcrystalline powder.
It is freely soluble in water, and practically insoluble inethanol (99.5).
Change the Identiˆcation to read:
Identiˆcation Determine the infrared absorption spectrumof Tranexamic Acid as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Tranexamic Acid Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.
Add the following next to Identiˆcation:
pH The pH of a solution prepared by dissolving 1.0 g ofTranexamic Acid in 20 mL of water is between 7.0 and 8.0.
Change the Purity to read:
Purity (1) Clarity and color of solution—Dissolve 1.0 gof Tranexamic Acid in 10 mL of water: the solution is clearand colorless.
(2) Chloride—Perform the test with 1.0 g of TranexamicAcid. Prepare the control solution with 0.40 mL of 0.01mol/L hydrochloric acid VS (not more than 0.014z).
(3) Heavy metals—Dissolve 2.0 g of Tranexamic Acid inwater to make 20 mL, and use this solution as the samplestock solution. To 12 mL of the sample stock solution add2 mL of hydrochloric acid-ammonium acetate buŠer solu-tion, pH 3.5, mix, add 1.2 mL of thioacetamide TS , miximmediately, and use this solution as the sample solution.Separately, proceed in the same manner as above with amixture of 1 mL of Standard Lead Solution, 2 mL of thesample stock solution and 9 mL of water, and use the solu-tion so obtained as the standard solution. Separately, pro-ceed in the same manner with a mixture of 10 mL of waterand 2 mL of the sample stock solution, and use the solutionso obtained as the control solution. Conform that the colorof the standard solution is slightly darker than that of the
control solution. Compare the sample solution and the stan-dard solution 2 minutes after they are prepared: the color ofthe sample solution is not more intense than that of the stan-dard solution (not more than 10 ppm).
(4) Arsenic—Prepare the test solution by dissolving1.0 g of Tranexamic Acid in 10 mL of water, and performthe test (not more than 2 ppm).
(5) Related substances—Dissolve 0.20 g of TranexamicAcid in water to make exactly 20 mL, and use this solutionas the sample solution. Pipet 5 mL of the sample solution,and add water to make exactly 100 mL. Pipet 1 mL of thissolution, add water to make exactly 10 mL, and use thissolution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the area multi-plied by relative response factor 1.2 of the peak, having therelative retention time of about 1.5 with respect to tranexam-ic acid, is not more than 2/5 of the peak area of tranexamicacid from the standard solution, and the area of the peak,having the relative retention time of about 2.1 with respectto tranexamic acid, is not more than 1/5 of the peak area oftranexamic acid from the standard solution. The area ofeach peak other than tranexamic acid and other than thepeaks mentioned above is not more than 1/5 of the peakarea of tranexamic acid from the standard solution. For thiscomparison, use the area of the peaks, having the relativeretention time of about 1.1 and about 1.3, after multiplyingby their relative response factors 0.005 and 0.006, respec-tively. The total area of the peaks other than tranexamic acidis not more than the peak area of tranexamic acid from thestandard solution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 3 times as long as theretention time of tranexamic acid after the solvent peak.System suitability—
Test for required detectability: To exactly 5 mL of thestandard solution add water to make exactly 25 mL. Con-ˆrm that the peak area of tranexamic acid obtained from20 mL of this solution is equivalent to 14 to 26z of that from20 mL of the standard solution.
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tranexamic acid is not more than 7z.
Change the Assay to read:
Assay Weigh accurately about 50 mg each of TranexamicAcid and Tranexamic Acid Reference Standard, previouslydried, dissolve in water to make exactly 25 mL, and use thesesolutions as the sample solution and the standard solution.
17461746 Supplement II, JPXIVO‹cial Monographs for Part I
Perform the test with exactly 20 mL each of the samplesolution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and determine the peak areas, AT and AS, of tranex-amic acid.
Amount (mg) of C8H15NO2＝WS×AT
AS
WS: Amount (mg) of Tranexamic Acid ReferenceStandard
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 220 nm).Column: A stainless steel column 6.0 mm in inside
diameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: Dissolve 11.0 g of sodium dihydrogenphosphate in 500 mL of water, and add 5 mL of triethyla-mine and 1.4 g of sodium lauryl sulfate. Adjust the pH to2.5 with phosphoric acid or diluted phosphoric acid (1 in10), add water to make 600 mL, and add 400 mL ofmethanol.
Flow rate: Adjust the ‰ow rate so that the retention timeof tranexamic acid is about 20 minutes.System suitability—
System performance: To 5 mL of the standard solutionadd 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in10,000) and water to make 50 mL. When the procedure isrun with 20 mL of this solution under the above operatingconditions, tranexamic acid and 4-(aminomethyl)benzoicacid are eluted in this order with the resolution between thesepeaks being not less than 5.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tranexamic acid is not more than 0.6z.
Add the following:
Tranexamic Acid Capsulesトラネキサム酸カプセル
Tranexamic Acid Capsules contain not less than95.0z and not more than 105.0z of the labeledamount of tranexamic acid (C8H15NO2: 157.21).
Method of preparation Prepare as directed underCapsules, with Tranexamic Acid.
Identiˆcation Take an amount of powdered contents ofTranexamic Acid Capsules, equivalent to 0.5 g of Tranex-amic Acid according to the labeled amount, add 50 mL ofwater, shake well, and ˆlter. To 5 mL of the ˆltrate add1 mL of ninhydrin TS, and heat for 3 minutes: a dark purple
color develops.
Mass variation It meets the requirements of the MassVariation Test.
Dissolution Being speciˆed separately.
Assay Weigh accurately the mass of the contents of notless than 20 Tranexamic Acid Capsules, and powder. Weighaccurately an amount of the powder, equivalent to about 0.1g of tranexamic acid (C8H15NO2), add 30 mL of water, shakewell, and add water to make exactly 50 mL. Centrifuge, ˆlterthe supernatant liquid through a membrane ˆlter with poresize of not more than 0.45 mm, discard the ˆrst 10 mL of theˆltrate, and use the subsequent ˆltrate as the sample solu-tion. Separately, weigh accurately about 50 mg of Tranex-amic Acid Reference Standard, previously dried at 1059Cfor 2 hours, dissolve in water to make exactly 25 mL, anduse this solution as the standard solution. Perform the testwith exactly 30 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine thepeak areas, AT and AS, of tranexamic acid.
Amount (mg) of tranexamic acid (C8H15NO2)
＝WS×AT
AS×2
WS: Amount (mg) of Tranexamic Acid Reference Stan-dard
Operating conditions—Detector, column, and mobile phase: Proceed as directed
in the operating conditions in the Assay under TranexamicAcid.
Column temperature: A constant temperature of about359C.
Flow rate: Adjust the ‰ow rate so that the retention timeof tranexamic acid is about 16 minutes.System suitability—
System performance: To 5 mL of the standard solutionadd 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in10,000) and water to make 50 mL. When the procedure isrun with 30 mL of this solution under the above operatingconditions, tranexamic acid and 4-(aminomethyl)benzoicacid are eluted in this order with the resolution between thesepeaks being not less than 3.
System repeatability: When the test is repeated 6 timeswith 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tranexamic acid is not more than 1.0z.
Containers and storage Containers—Tight containers.
17471747Supplement II, JPXIV O‹cial Monographs for Part I
Add the following:
Tranexamic Acid Injectionトラネキサム酸注射液
Tranexamic Acid Injection is an aqueous injection.It contains not less than 95.0z and not more than
105.0z of the labeled amount of tranexamic acid(C8H15NO2: 157.21).
Method of preparation Prepare as directed underInjections, with Tranexamic Acid.
Description Tranexamic Acid Injection is a clear andcolorless liquid.
Identiˆcation To a volume of Tranexamic Acid Injection,equivalent to 50 mg of Tranexamic Acid according to thelabeled amount, add water to make 5 mL, add 1 mL ofninhydrin TS, and heat: a dark purple color develops.
pH 7.0 – 8.0
Bacterial endotoxins Not more than 0.12 EU/mg.
Actual volume It meets the requirements of Injections.
Foreign insoluble matter Perform the test according toMethod 1: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay Take accurately a volume of Tranexamic AcidInjection, equivalent to about 0.1 g of tranexamic acid(C8H15NO2), add water to make exactly 50 mL, and use thissolution as the sample solution. Separately, weigh accuratelyabout 50 mg of Tranexamic Acid Reference Standard,previously dried at 1059C for 2 hours, dissolve in water tomake exactly 25 mL, and use this solution as the standardsolution. Perform the test with exactly 30 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and determine the peak areas, AT and AS, oftranexamic acid.
Amount (mg) of tranexamic acid (C8H15NO2)＝WS×AT
AS
WS: Amount (mg) of Tranexamic Acid ReferenceStandard
Operating conditions—Detector, column, and mobile phase: Proceed as directed
in the operating conditions in the Assay under TranexamicAcid.
Column temperature: A constant temperature of about
359C.Flow rate: Adjust the ‰ow rate so that the retention time
of tranexamic acid is about 16 minutes.System suitability—
System performance: To 5 mL of the standard solutionadd 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in10,000) and water to make 50 mL. When the procedure isrun with 30 mL of this solution under the above operatingconditions, tranexamic acid and 4-(aminomethyl)benzoicacid are eluted in this order with the resolution between thesepeaks being not less than 3.
System repeatability: When the test is repeated 6 timeswith 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tranexamic acid is not more than 1.0z.
Containers and storage Containers—Hermetic containers.
Add the following:
Tranexamic Acid Tabletsトラネキサム酸錠
Tranexamic Acid Tablets contain not less than95.0z and not more than 105.0z of the labeledamount of tranexamic acid (C8H15NO2: 157.21).
Method of preparation Prepare as directed under Tablets,with Tranexamic Acid.
Identiˆcation To an amount of powdered TranexamicAcid Tablets, equivalent to 0.5 g of Tranexamic Acidaccording to the labeled amount, add 50 mL of water, shakewell, and ˆlter. To 5 mL of the ˆltrate add 1 mL ofninhydrin TS, and heat for 3 minutes: a dark purple colordevelops.
Mass variation It meets the requirements of the MassVariation Test.
Dissolution Being speciˆed separately.
Assay Weigh accurately the mass of not less than 20Tranexamic Acid Tablets, and powder. Weigh accurately aquantity of the powder, equivalent to about 5 g of tranexam-ic acid (C8H15NO2), add 150 mL of water, disintegrate thetablets completely with the aid of ultrasonic waves, and addwater to make exactly 200 mL. Centrifuge, pipet 4 mL of thesupernatant liquid, and add water to make exactly 50 mL.Filter through a membrane ˆlter with pore size of not morethan 0.45 mm, discard the ˆrst 10 mL of the ˆltrate, and usethe subsequent ˆltrate as the sample solution. Separately,weigh accurately about 50 mg of Tranexamic Acid ReferenceStandard, previously dried at 1059C for 2 hours, dissolve inwater to make exactly 25 mL, and use this solution as thestandard solution. Perform the test with exactly 30 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the peak areas, AT and AS, of
17481748 Supplement II, JPXIVO‹cial Monographs for Part I
tranexamic acid.
Amount (mg) of tranexamic acid (C8H15NO2)
＝WS×AT
AS×100
WS: Amount (mg) of Tranexamic Acid ReferenceStandard
Operating conditions—Detector, column, and mobile phase: Proceed as directed
in the operating conditions in the Assay under TranexamicAcid.
Column temperature: A constant temperature of about359C.
Flow rate: Adjust the ‰ow rate so that the retention timeof tranexamic acid is about 16 minutes.System suitability—
System performance: To 5 mL of the standard solutionadd 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in10,000) and water to make 50 mL. When the procedure isrun with 30 mL of this solution under the above operatingconditions, tranexamic acid and 4-(aminomethyl)benzoicacid are eluted in this order with the resolution between thesepeaks being not less than 3.
System repeatability: When the test is repeated 6 timeswith 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tranexamic acid is not more than 1.0z.
Containers and storage Containers—Tight containers.
Trichlormethiazideトリクロルメチアジド
Change the origin/limits of the content to read:
Trichlormethiazide, when dried, contains not lessthan 97.5z and not more than 102.0z ofC8H8Cl3N3O4S2.
Change the Description to read:
Description Trichlormethiazide occurs as a white powder.It is freely soluble in N,N-dimethylformamide and in
acetone, slightly soluble in acetonitrile and in ethanol (95),and practically insoluble in water.
A solution of Trichlormethiazide in acetone (1 in 50)shows no optical rotation.
Melting point: about 2709C (with decomposition).
Change the Identiˆcation to read:
Identiˆcation(1) Determine the absorption spectrum of a solution of
Trichlormethiazide in ethanol (95) (3 in 250,000) as directedunder the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of a solution of Trichlormethiazide Reference
Standard prepared in the same manner as the samplesolution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths.
(2) Determine the infrared absorption spectrum ofTrichlormethiazide as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Trichlormethiazide Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.
(3) Perform the test with Trichlormethiazide as directedunder the Flame Coloration Test (2): a green color appears.
Change the Purity (4) and (5) to read:
(4) Arsenic—Prepare the test solution with 0.6 g ofTrichlormethiazide according to Method 5, using 20 mL ofN,N-dimethylformamide, and perform the test (not morethan 3.3 ppm).
(5) Related substances―Dissolve 25 mg of Trichlor-methiazide in 50 mL of acetonitrile, and use the solution asthe sample solution. Perform the test with 10 mL of the sam-ple solution as directed under the Liquid Chromatographyaccording to the following conditions, determine each peakarea by the automatic integration method, and calculate theamount of related substances by the area percentagemethod: the amount of 4-amino-6-chlorobenzene-1,3-disul-fonamide, having the relative retention time of about 0.3with respect to trichlormethiazide, is not more than 2.0z,and the total amount of the related substances is not morethan 2.5z.Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 268 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with phenylsilanizedsilica gel for liquid chromatography (5 mm in particlediameter).
Column temperature: A constant temperature of about259C.
Mobile phase A: A mixture of diluted phosphoric acid(1 in 1000) and acetonitrile (3:1).
Mobile phase B: A mixture of acetonitrile and dilutedphosphoric acid (1 in 1000) (3:1).
Flowing of the mobile phase: Control the gradient bymixing the mobile phases A and B as directed in the follow-ing table.
Time after injectionof sample (min)
Mobile phaseA (volz)
Mobile phaseB (volz)
0 – 10 100 010 – 20 100 ª 0 0 ª 100
Flow rate: 1.5 mL per minuteTime span of measurement: About 2.5 times as long as the
retention time of trichlormethiazide after the solvent peak.System suitability—
Test for required detectability: To exactly 1 mL of the
17491749Supplement II, JPXIV O‹cial Monographs for Part I
sample solution add acetonitrile to make exactly 50 mL, anduse this solution as the solution for system suitability test.Pipet 1 mL of the solution, and add acetonitrile to make ex-actly 20 mL. Conˆrm that the peak area of trichlor-methiazide obtained from 10 mL of this solution is equiva-lent to 3.5 to 6.5z of that of trichlormethiazide obtainedfrom 10 mL of the solution for system suitability test.
System performance: To 5 mL of the solution for systemsuitability test add 5 mL of water, and warm in a water bathat 609C for 30 minutes. When the procedure is run with10 mL of this solution, after cooling, under the above operat-ing conditions, 4-amino-6-chlorobenzene-1,3-disulfonamideand trichlormethiazide are eluted in this order, the relativeretention time of 4-amino-6-chlorobenzene-1,3-disul-fonamide with respect to trichlormethiazide is about 0.3,and the number of theoretical plates and the symmetry fac-tor of the peak of trichlormethiazide are not less than 5000and not more than 1.2, respectively.
System repeatability: When the test is repeated 3 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of trichlormethiazide is not more than2.0z.
Change the Assay to read:
Assay Weigh accurately about 25 mg of Trichlor-methiazide and Trichlormethiazide Reference Standard,previously dried, and dissolve separately in exactly 20 mL ofthe internal standard solution. To 1 mL of these solutionsadd acetonitrile to make 20 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of trichlormethiazide tothat of the internal standard.
Amount (mg) of C8H8Cl3N3O4S2＝WS×QT
QS
WS: Amount (mg) of Trichlormethiazide Reference Stan-dard
Internal standard solution—A solution of 3-nitrophenol inacetonitrile (1 in 800).Operating conditions—
Detector: An ultraviolet absorption photometer(wavelength: 268 nm).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with phenylsilanizedsilica gel for liquid chromatography (5 mm in particlediameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of diluted phosphoric acid (1 in1000) and acetonitrile (3:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof trichlormethiazide is about 8 minutes.System suitability—
System performance: When the procedure is run with
10 mL of the standard solution under the above operatingconditions, the internal standard and trichlormethiazide areeluted in this order with the resolution between these peaksbeing not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of trichlormethiazide to that of the internalstandard is not more than 1.0z.
Add the following:
Trichlormethiazide Tabletsトリクロルメチアジド錠
Trichlormethiazide Tablets contain not less than93.0z and not more than 107.0z of the labeledamount of trichlormethiazide (C8H8Cl3N3O4S2:380.66).
Method of preparation Prepare as directed under Tablets,with Trichlormethiazide.
Identiˆcation To an amount of pulverized Trichlor-methiazide Tablets, equivalent to 4 mg of Trichlor-methiazide according to the labeled amount, add 10 mL ofacetone, shake vigorously for 5 minutes, centrifuge, and usethe supernatant liquid as the sample solution. Separately,dissolve 4 mg of Trichlormethiazide Reference Standard in10 mL of acetone, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 5 mL each ofthe sample solution and the standard solution on a plate ofsilica gel with ‰uorescent indicator for thin-layer chro-matography. Develop the plate with a mixture of ethylacetate, hexane and methanol (10:4:1) to a distance of about10 cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 254 nm): the principal spots from thesample solution and the standard solution show the same Rfvalue.
Purity Related substances—Pulverize a suitable amount ofTrichlormethiazide Tablets in an agate mortar. Take anamount of the powder, equivalent to 10 mg of Trichlor-methiazide according to the labeled amount, add 20 mL ofacetonitrile, shake vigorously for 15 minutes, centrifuge,and use the supernatant liquid as the sample solution. Per-form the test with 10 mL of the sample solution as directedunder the Liquid Chromatography according to the follow-ing conditions, determine each peak area by the automaticintegration method, and calculate the amount of eachrelated substance by the area percentage method: theamount of 4-amino-6-chlorobenzene-1,3-disulfoneamide,having the relative retention time of about 0.3 with respectto trichlormethiazide, is not more than 4.0z, and the totalamount of the peaks other than trichlormethiazide is notmore than 5.0z.
17501750 Supplement II, JPXIVO‹cial Monographs for Part I
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 268 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 15 cm in length, packed with phenylsilanizedsilica gel for liquid chromatography (5 mm in particlediameter).
Column temperature: A constant temperature of about259C.
Mobile phase A: A mixture of diluted phosphoric acid(1 in 1000) and acetonitrile (3:1).
Mobile phase B: A mixture of acetonitrile and dilutedphosphoric acid (1 in 1000) (3:1).
Flowing of the mobile phase: Control the gradient bymixing the mobile phases A and B as directed in the follow-ing table.
Time after injectionof sample (min)
Mobile phaseA (z)
Mobile phaseB (z)
0 – 10 100 010 – 20 100 ª 0 0 ª 100
Flow rate: 1.5 mL/minuteTime span of measurement: About 2.5 times as long as the
retention time of trichlormethiazide after the solvent peak.System suitability—
Test for required detectability: Dissolve 25 mg ofTrichlormethiazide in 50 mL of acetonitrile. Pipet 1 mL ofthis solution, add acetonitrile to make exactly 50 mL, anduse this solution as the solution for system suitability test.Pipet 1 mL of the solution for system suitability test, andadd acetonitrile to make exactly 20 mL. Conˆrm that thepeak area of trichlormethiazide obtained from 10 mL of thissolution is equivalent to 3.5 to 6.5z of that obtained from10 mL of the solution for system suitability test.
System performance: To 5 mL of the solution for systemsuitability test add 5 mL of water, and warm in a water bathof 609C for 30 minutes. When the procedure is run with 10mL of this solution, after cooling, under the above operatingconditions, 4-amino-6-chlorobenzene-1,3-disulfonamideand trichlormethiazide are eluted in this order, the relativeretention time of 4-amino-6-chlorobenzene-1,3-disul-fonamide with respect to trichlormethiazide is about 0.3,and the number of theoretical plates and the symmetry fac-tor of the peak of trichlormethiazide are not less than 5000and not more than 1.2, respectively.
System repeatability: When the test is repeated 3 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of trichlormethiazide is not more than2.0z.
Content uniformity Perform the test according to the fol-lowing method: it meets the requirements of the ContentUniformity Test. To one tablet of TrichlormethiazideTablets add 5 mL of diluted phosphoric acid (1 in 50) to dis-integrate. Add exactly an amount of the internal standardsolution, equivalent to 10 mL per 2 mg of trichlormethiazide
(C8H8Cl3N3O4S2) according to the labeled amount, addacetonitrile to make 25 mL, shake vigorously for 15minutes, and centrifuge. To an amount of the supernatantliquid add the mobile phase to make a solution so that it con-tains about 40 mg of trichlormethiazide (C8H8Cl3N3O4S2) ineach mL, and use this solution as the sample solution.Separately, weigh accurately about 20 mg of Trichlor-methiazide Reference Standard, previously dried at 1059Cfor 3 hours, and dissolve in acetonitrile to make exactly 100mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add 10 mL of acetonitrile and 5 mLof diluted phosphoric acid (1 in 50), and use this solution asthe standard solution. Perform the test with 20 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the conditionsdescribed in Assay, and determine the ratios, QT and QS, ofthe peak area of trichlormethiazide to that of the internalstandard.
Amount (mg) of trichlormethiazide (C8H8Cl3N3O4S2)
＝WS×QT
QS×C×
120
WS: Amount (mg) of Trichlormethiazide ReferenceStandard
C: Labeled amount (mg) of trichlormethiazide(C8H8Cl3N3O4S2) per tablet
Internal standard solution—A solution of 3-nitrophenol inacetonitrile (1 in 5000).
Dissolution Perform the test with 1 tablet of Trichlor-methiazide Tablets at 50 revolutions per minute according toMethod 2 under the Dissolution Test, using 900 mL of wateras the dissolution medium. Withdraw 20 mL or more of thedissolution medium 15 minutes after starting the test, andˆlter through a membrane ˆlter with pore size of not morethan 0.45 mm. Discard the ˆrst 10 mL of the ˆltrate, pipetthe subsequent V mL of the ˆltrate, add diluted phosphoricacid (1 in 50) to make exactly V? mL so that each mL con-tains about 1.1 mg of trichlormethiazide (C8H8Cl3N3O4S2)according to the labeled amount, and use this solution as thesample solution. Separately, weigh accurately about 22 mgof Trichlormethiazide Reference Standard, previously driedat 1059C for 3 hours, and dissolve in acetonitrile to make ex-actly 200 mL. Pipet 2 mL of this solution, add diluted phos-phoric acid (1 in 50) to make exactly 200 mL, and use this so-lution as the standard solution. Perform the test with exactly40 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine the peak areas, ATa
and ASa, of trichlormethiazide obtained with the sample so-lution and the standard solution, and the area, ATb, of thepeak, having the relative retention time of about 0.3 withrespect to trichlormethiazide, obtained with the sample solu-tion. The dissolution rate in 15 minutes is not less than 75z.
Dissolution rate (z) with respect to the labeled amount oftrichlormethiazide (C8H8Cl3N3O4S2)
17511751Supplement II, JPXIV O‹cial Monographs for Part I
＝WS×ATa＋ATb×0.95
ASa×
V?V×
1C×
92
WS: Amount (mg) of Trichlormethiazide Reference Stan-dard
C: Labeled amount (mg) of trichlormethiazide(C8H8Cl3N3O4S2) per tablet
Operating conditions—Proceed as directed in the operating conditions in the
Assay.System suitability—
System performance: Dissolve 25 mg of Trichlor-methiazide in 50 mL of acetonitrile. To 1 mL of this solutionadd acetonitrile to make 50 mL. To 5 mL of this solutionadd 5 mL of water, and heat at 609C in a water bath for 30minutes. After cooling, when the procedure is run with10 mL of this solution under the above operating conditions,4-amino-6-chlorobenzene-1,3-disulfonamide and trichlor-methiazide are eluted in this order, the relative retentiontime of 4-amino-6-chlorobenzene-1,3-disulfonamide withrespect to trichlormethiazide is about 0.3, and the number oftheoretical plates and the symmetry factor of the peak oftrichlormethiazide are not less than 5000 and not more than1.2, respectively.
System repeatability: When the test is repeated 6 timeswith 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of trichlormethiazide is not more than 2.0z.
Assay Weigh accurately the mass of not less than 20Trichlormethiazide Tablets, and pulverize the tablets in anagate mortar. Weigh accurately an amount of the powder,equivalent to about 2 mg of trichlormethiazide(C8H8Cl3N3O4S2) according to the labeled amount, add 5 mLof diluted phosphoric acid (1 in 50) and exactly 10 mL of theinternal standard solution, add 10 mL of acetonitrile, shakevigorously for 15 minutes, and centrifuge. To 2 mL of thesupernatant liquid add 2 mL of the mobile phase, and usethis solution as the sample solution. Separately, weigh ac-curately about 40 mg of Trichlormethiazide Reference Stan-dard, previously dried at 1059C for 3 hours, and dissolve inacetonitrile to make exactly 200 mL. Pipet 5 mL of this solu-tion, add exactly 5 mL of the internal standard solution, add10 mL of acetonitrile and 5 mL of diluted phosphoric acid (1in 50), and use this solution as the standard solution. Per-form the test with 20 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine the ratios, QT and QS, of the peak area of trichlor-methiazide to that of the internal standard.
Amount (mg) of trichlormethiazide (C8H8Cl3N3O4S2)
＝WS×QT
QS×
120
WS: Amount (mg) of Trichlormethiazide ReferenceStandard
Internal standard solution—A solution of 3-nitrophenol inacetonitrile (1 in 5000).
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 268 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 15 cm in length, packed with phenylsilanizedsilica gel for liquid chromatography (5 mm in particlediameter).
Column temperature: A constant temperature of about259C.
Mobile phase: A mixture of diluted phosphoric acid (1 in1000) and acetonitrile (3:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof trichlormethiazide is about 8 minutes.System suitability—
System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, the internal standard and trichlormethiazide areeluted in this order with the resolution between these peaksbeing not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of trichlormethiazide to that of the internalstandard is not more than 1.0z.
Containers and storage Containers—Tight containers.
Vasopressin Injectionバソプレシン注射液
Change the paragraph (i) under Purity to read:
Purity(i) Standard stock solution: Dissolve 200 oxytocin Units
of Posterior Pituitary Reference Standard, according to thelabeled Units, in exactly 10 mL of diluted acetic acid (100)(1 in 400). Pipet 1 mL of this solution, and add diluted aceticacid (100) (1 in 400) to make exactly 10 mL. Store in a coldplace, avoiding freezing. Use within 6 months from the dateof preparation.
Change the paragraph (ii) under Assay to read:
Assay(ii) Standard stock solution: Dissolve 2000 vasopressin
Units of Posterior Pituitary Reference Standard, accordingto the labeled Units, in exactly 100 mL of diluted acetic acid(100) (1 in 400). Pipet 1 mL of this solution, and add dilutedacetic acid (100) (1 in 400) to make exactly 10 mL. Store in acold place, avoiding freezing. Use within 6 months from thedate of preparation.
17521752 Supplement II, JPXIVO‹cial Monographs for Part I
Vinblastine Sulfate硫酸ビンブラスチン
Change to read except the structural formula,chemical name and origin/limits of content:
Description Vinblastine Sulfate occurs as a white to paleyellow powder.
It is soluble in water, sparingly soluble in methanol, andpractically insoluble in ethanol (99.5).
It is hygroscopic.Optical rotation [a]D20: －28 –－359(0.20 g calculated on
the dried basis, methanol, 10 mL, 100 mm).
Identiˆcation (1) Determine the absorption spectrum ofa solution of Vinblastine Sulfate (1 in 50,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or thespectrum of a solution of Vinblastine Sulfate ReferenceStandard prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption atthe same wavelengths.
(2) Determine the infrared absorption spectrum ofVinblastine Sulfate as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Vinblastine Sulfate Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.
(3) A solution of Vinblastine Sulfate (1 in 100) respondsto the Qualitative Tests for sulfate.
pH Dissolve 15 mg of Vinblastine Sulfate in 10 mL ofwater: the pH of this solution is between 3.5 and 5.0.
Purity (1) Clarity and color of solution—Dissolve 50 mgof Vinblastine Sulfate in 10 mL of water: the solution is clearand colorless.
(2) Related substances—Dissolve about 4 mg of Vin-blastine Sulfate in 10 mL of water, and use this solution asthe sample solution. Pipet 1 mL of the sample solution, addwater to make exactly 25 mL, and use this solution as thestandard solution. Perform the test with exactly 200 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area of these solu-tions by the automatic integration method: the area of peakother than the main peak is not larger than 1/4 of the peakarea of vinblastine from the standard solution, and the totalarea of the peaks other than the main peak is not larger than3/4 of the peak area of vinblastine from the standardsolution.Operating conditions—
Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.
Time span of measurement: About 4 times as long as theretention time of vinblastine after the solvent peak.
System suitability—Test for required detectability: To exactly 2.5 mL of the
standard solution add water to make exactly 100 mL.Conˆrm that the peak area of vinblastine obtained from200 mL of this solution is equivalent to 1.7 to 3.3z of thatfrom 200 mL of the standard solution.
System performance: Proceed as directed in the systemsuitability in the Assay.
System repeatability: When the test is repeated 5 timeswith 200 mL of the standard solution under the aboveoperating conditions, the relative standard deviation of thepeak area of vinblastine is not more than 1.5z.
Loss on drying Perform the test with about 10 mg ofVinblastine Sulfate as directed in Method 2 under the Ther-mal Analysis according to the following conditions: notmore than 15.0z.Operating conditions—
Heating rate: 59C/minuteTemperature range: room temperature to 2009CAtmospheric gas: dried NitrogenFlow rate of atmospheric gas: 40 mL/minute
Assay Weigh accurately about 10 mg each of VinblastineSulfate and Vinblastine Sulfate Reference Standard(separately determine its loss on drying), dissolve in water tomake exactly 25 mL, and use these solutions as the samplesolution and the standard solution, respectively. Performthe test with exactly 20 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine the peak areas, AT and AS, of vinblastine.
Amount (mg) of C46H58N4O9.H2SO4＝WS×AT
AS
WS: Amount (mg) of Vinblastine Sulfate ReferenceStandard, calculated on the dried basis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 262 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about259C.
Mobile phase: To 7 mL of diethylamine add water tomake 500 mL, and adjust the pH to 7.5 with phosphoricacid. To 380 mL of this solution add 620 mL of a mixture ofmethanol and acetonitrile (4:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof vinblastine is about 8 minutes.System suitability—
System performance: Dissolve 10 mg each of VinblastineSulfate and vincristine sulfate in 25 mL of water. When theprocedure is run with 20 mL of this solution under the aboveoperating conditions, vincristine and vinblastine are elutedin this order with the resolution between these peaks being
17531753Supplement II, JPXIV O‹cial Monographs for Part I
not less than 4.System repeatability: When the test is repeated 5 times
with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of vinblastine is not more than 1.0z.
Containers and storage Containers—Tight containers.Storage—Light-resistant, at not exceeding －209C.
Change to read:
Vinblastine Sulfate for Injection注射用硫酸ビンブラスチン
Vinblastine Sulfate for Injection is a preparation forinjection, which is dissolved before use. It containsnot less than 90.0z and not more than 110.0z ofthe labeled amount of vinblastine sulfate(C46H58N4O9.H2SO4: 909.05).
Method of preparation Prepare as directed under Injec-tions, with Vinblastine Sulfate.
Description Vinblastine Sulfate for Injection occurs aswhite to pale yellow, light masses or powder.
It is freely soluble in water.The pH of a solution (1 in 1000) is 3.5 – 5.0.
Identiˆcation Proceed as directed in the Identiˆcation (1)under Vinblastine Sulfate.
Purity Related substances—Dissolve 4 mg of VinblastineSulfate for Injection in 10 mL of water, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add water to make exactly 25 mL, and use this solution asthe standard solution. Perform the test with exactly 200 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the area of the peak otherthan the main peak from the sample solution is not largerthan 1/2 of the peak area of vinblastine from the standardsolution, and the total area of the peaks other than the mainpeak is not larger than 2 times the peak area of vinblastinefrom the standard solution.Operating conditions—
Perform as directed in the operating conditions in Purity(2) under Vinblastine Sulfate.System suitability—
Perform as directed in the system suitability in Purity (2)under Vinblastine Sulfate.
Bacterial endotoxins Less than 10 EU/mg.
Content uniformity Perform the test according to the fol-
lowing method: it meets the requirements of the ContentUniformity Test. Open a container of Vinblastine Sulfatefor Injection, add an exact amount of water to make a solu-tion so that each mL contains about 0.4 mg of vinblastinesulfate (C46H58N4O9.H2SO4) according to the labeledamount, and use this solution as the sample solution.Separately, weigh accurately about 10 mg of VinblastineSulfate Reference Standard (separately determine its loss ondrying in the same manner as for Vinblastine Sulfate), dis-solve in water to make exactly 25 mL, and use this solutionas the standard solution. Proceed as directed in the Assayunder Vinblastine Sulfate.
Amount (mg) of vinblastine sulfate (C46H58N4O9.H2SO4)
＝WS×AT
AS
WS: Amount (mg) of Vinblastine Sulfate ReferenceStandard, calculated on the dried basis
Foreign insoluble matter Perform the test according toMethod 2: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Assay Take an amount of Vinblastine Sulfate for Injec-tion, equivalent to 0.10 g of vinblastine sulfate(C46H58N4O9.H2SO4), dissolve each content with a suitableamount of water, transfer into a 100-mL volumetric ‰ask,wash each container with water, transfer the washings intothe volumetric ‰ask, and add water to make exactly 100 mL.Pipet 10 mL of this solution, add water to make exactly 25mL, and use this solution as the sample solution. Separately,weigh accurately about 10 mg of Vinblastine Sulfate Refer-ence Standard (separately determine its loss on drying in thesame manner as for Vinblastine Sulfate), dissolve in water tomake exactly 25 mL, and use this solution as the standardsolution. Proceed as directed in the Assay under VinblastineSulfate.
Amount (mg) of vinblastine sulfate (C46H58N4O9.H2SO4)
＝WS×AT
AS
WS: Amount (mg) of Vinblastine Sulfate ReferenceStandard, calculated on the dried basis
Containers and storage Containers—Hermetic containers,and colored containers may be used.
Storage—Light-resistant, at 2 to 89C.
17551755
General Rulesfor Crude Drugs
Change the paragraph 1 to read:
1. Crude drugs in the monographs include medici-nal parts obtained from plants or animals, cell inclu-sions and secretes separated from the origins, theirextracts, and minerals. General Rules for Crude Drugsand the Crude Drugs in General Tests, Processes andApparatus are applicable to the followings:
Powdered Aloe is the powder of Aloe.It contains not less than 4.0z of barbaloin, calcu-
lated on the basis of dried material.
Change the Loss on drying to read:
Loss on drying Not more than 12.0z.
Add the following next to Extract content:
Component determination Weigh accurately about 0.1 gof Powdered Aloe, add 40 mL of methanol, and heat undera re‰ex condenser on a water bath for 30 minutes. Aftercooling, ˆlter, and add methanol to the ˆltrate to makeexactly 50 mL. Pipet 5 mL of the solution, add methanol tomake exactly 10 mL, and use this solution as the samplesolution. Separately, weigh accurately about 10 mg ofbarbaloin for component determination, previously dried ina desiccator (in vacuum, phosphorus (V) oxide) for 24hours, add 40 mg of oxalic acid dihydrate, and dissolve inmethanol to make exactly 100 mL. Pipet 5 mL of thesolution, add methanol to make exactly 10 mL, and use thissolution as the standard solution. Perform the test withexactly 5 mL each of the sample solution and the standard
solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine thepeak areas of barbaloin, AT and AS, of both solutions.
Amount (mg) of barbaloin＝WS×AT
AS×2
WS: Amount (mg) of barbaloin for component determina-tion
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 360 nm).Column: A stainless steel column about 6 mm in inside
diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).
Column temperature: A constant temperature of about309C.
Mobile phase: A mixture of water, acetonitrile and aceticacid (100) (74:26:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof barbaloin is about 12 minutes.System suitability—
System performance: To about 10 mg of barbaloin forcomponent determination add 40 mg of oxalic acid dihy-drate, and dissolve in methanol to make exactly 100 mL.Pipet 5 mL of the solution, add 1 mL of a solution of ethen-zamide in methanol (1 in 2000) and methanol to makeexactly 10 mL. When the procedure is run with 5 mL of thissolution under the above operating conditions except thewavelength of 300 nm, barbaloin and ethenzamide areeluted in this order with the resolution between these peaksbeing not less than 2.0.
System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of barbaloin is not more than 1.5z.
Benzyl Alcoholベンジルアルコール
Change to read except the structural formulaand chemical name:
Benzyl Alcohol contains not less than 98.0z andnot more than 100.5z of C7H8O.
The label states, where applicable, that it is suitablefor use in the manufacture of injection forms.
17581758 Supplement II, JPXIVO‹cial Monographs for Part II
Description Benzyl Alcohol is a clear, colorless oily liquid.It is miscible with ethanol (95), with fatty oils and with
volatile oils.It is soluble in water.Speciˆc gravity d 20
20: 1.043 – 1.049
Identiˆcation Determine the infrared absorption spectrumof Benzyl Alcohol as directed in the liquid ˆlm methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.
Refractive index nD20: 1.538 – 1.541
Purity (1) Clarity and color of solution—Dissolve 2.0mL of Benzyl Alcohol in 60 mL of water: the solution isclear and colorless.
(2) Acid—To 10 mL of Benzyl Alcohol add 10 mL ofneutralized ethanol, 2 drops of phenolphthalein TS and 1.0mL of 0.1 mol/L sodium hydroxide VS: the color of thesolution is red.
(3) Benzaldehyde and other related substances—UseBenzyl Alcohol as the sample solution. Separately, weighexactly 0.750 g of benzaldehyde and 0.500 g of cyclohex-ylmethanol, and add Benzyl Alcohol to make exactly 25 mL.Pipet 1 mL of this solution, add exactly 2 mL of ethylben-zene internal standard solution and exactly 3 mL of dicyclo-hexyl internal standard solution, then add Benzyl Alcohol tomake exactly 20 mL, and use this solution as the standardsolution (1). Perform the test with 0.1 mL each of the samplesolution and the standard solution (1) as directed under theGas Chromatography according to the following conditions:no peaks of ethylbenzene and dicyclohexyl appear on thechromatogram obtained with the sample solution. When0.1 mL of the standard solution (1) is injected, adjust thesensitivity of the detector so that the peak height of ethyl-benzene is not more than 30z of the full scale of the record-er. The peak area of benzaldehyde obtained with the samplesolution is not more than the deference between the peakareas of benzaldehyde of the sample solution and thestandard solution (1) (0.15z), and the peak area of cyclo-hexylmethanol with the sample solution is not more than thedeference between the peak areas of cyclohexylmethanol ofthe sample solution and the standard solution (1) (0.10z).The total area of the peaks having smaller retention timethan benzyl alcohol and other than benzaldehyde andcyclohexylmethanol obtained with the sample solution is notmore than 4 times the peak area of ethylbenzene with thestandard solution (1) (0.04z). The total area of the peakshaving larger retention time than benzaldehyde obtainedwith the sample solution is not more than the peak area ofcyclohexylmethanol with the standard solution (1) (0.3z).For these calculations the peak areas less than 1/100 timesthe peak area of ethylbenzene with the standard solution (1)are excluded.
Benzyl Alcohol labeled that it is suitable for use in themanufacture of injection forms meets the following require-ments.
Use Benzyl Alcohol as the sample solution. Separately,
weigh exactly 0.250 g of benzaldehyde and 0.500 g of cyclo-hexylmethanol, and add Benzyl Alcohol to make exactly25 mL. Pipet 1 mL of this solution, add exactly 2 mL of theethylbenzene internal standard solution and exactly 2 mL ofthe dicyclohexyl internal standard solution, then add BenzylAlcohol to make exactly 20 mL, and use this solution as thestandard solution (2). Perform the test with 0.1 mL each ofthe sample solution and the standard solution (2) as directedunder the Gas Chromatography according to the followingconditions: no peaks of ethylbenzene and dicyclohexylappear on the chromatogram obtained with the samplesolution. When 0.1 mL of the standard solution (2) is inject-ed, adjust the sensitivity of the detector so that the peakheight of ethylbenzene is not more than 30z of the full scaleof the recorder. The peak area of benzaldehyde of obtainedwith the sample solution is not more than the deferencebetween the peak areas of benzaldehyde of the samplesolution and the standard solution (2) (0.05z), and the peakarea of cyclohexylmethanol with the sample solution is notmore than the deference between the peak areas of cyclohex-ylmethanol of the sample solution and the standard solution(2) (0.10z). The total area of the peaks having smallerretention time than benzyl alcohol and other than benzalde-hyde and cyclohexylmethanol obtained with the samplesolution is not more than 2 times the peak area of ethylben-zene with the standard solution (2) (0.02z). The total areaof the peaks having larger retention time than benzaldehydeobtained with the sample solution is not more than the peakarea of cyclohexylmethanol with the standard solution (2)(0.2z). For these calculation the peak areas less than 1/100times the peak area of ethylbenzene with the standardsolution (2) are excluded.
Ethylbenzene internal standard solution: Dissolve exactly0.100 g of ethylbenzene in Benzyl Alcohol to make exactly10 mL. Pipet 2 mL of this solution, and add Benzyl Alcoholto make exactly 20 mL.
Dicyclohexyl internal standard solution: Dissolve exactly2.000 g of dicyclohexyl in Benzyl Alcohol to make exactly10 mL. Pipet 2 mL of this solution, and add Benzyl Alcoholto make exactly 20 mL.Operating conditions—
Detector: A hydrogen ‰ame-ionization detectorColumn: A fused silica column 0.33 mm in inside
diameter and 30 m in length, coated inside with polyethyleneglycol 20M for gas chromatography in 0.5 mm thickness.
Column temperature: Raise the temperature at a rate of59C per minutes from 509C to 2209C, and maintain at2209C for 35 minutes.
Temperature of injection port: A constant temperature ofabout 2009C.
Temperature of detector: A constant temperature ofabout 3109C.
Carrier gas: HeliumFlow rate: Adjust the ‰ow rate so that the retention time
of benzyl alcohol is between 24 and 28 minutes.Split ratio: Splitless
System suitability—System performance: When the procedure is run with the
17591759Supplement II, JPXIV O‹cial Monographs for Part II
standard solution (1) under the above operating conditions,the relative retention times of ethylbenzene, dicyclohexyl,benzaldehyde and cyclohexylmethanol with respect to benzylalcohol are about 0.28, about 0.59, about 0.68 and about0.71, respectively, and the resolution between the peaks ofbenzaldehyde and cyclohexylmethanol is not less than 3.0.In the case of Benzyl Alcohol labeled to use for injection,proceed with the standard solution (2) instead of thestandard solution (1).
(4) Peroxide value—Dissolve 5 g of Benzyl Alcohol in30 mL of a mixture of acetic acid (100) and chloroform (3:2)in a 250-mL glass-stoppered conical ‰ask. Add 0.5 mL ofpotassium iodide saturated solution, shake exactly for 1minute, add 30 mL of water, and titrate with 0.01 mol/Lsodium thiosulfate VS until the blue color of the solutiondisappears after addition of 10 mL of starch TS near the endpoint where the solution is a pale yellow color. Perform ablank determination in the same manner. Calculate theamount of peroxide by the following formula: not morethan 5.
Amount (mEq/kg) of peroxide＝(V1－V0)
W×10
V1: Volume (mL) of 0.01 mol/L sodium thiosulfate VSconsumed in the test
V0: Volume (mL) of 0.01 mol/L sodium thiosulfate VSconsumed in the blank
W: Amount (g) of the sample
(5) Residue on evaporation—Perform the test afterconformation that the sample meets the requirement of theperoxide value. Evaporate to dryness 10.0 g of BenzylAlcohol on a water bath, dry the residue at 1059C for 1hour, and allow to cool in a desiccator: not more than 5 mg.
Assay Weigh accurately about 0.9 g of Benzyl Alcohol,add exactly 15.0 mL of a mixture of pyridine and aceticanhydride (7:1), and heat on a water bath under a re‰uxcondenser for 30 minutes. Cool, add 25 mL of water, andtitrate the excess acetic acid with 1 mol/L sodium hydroxideVS (indicator: 2 drops of phenolphthalein TS). Perform ablank determination.
Each mL of 1 mol/L sodium hydroxide VS＝108.1 mg of C7H8O
Containers and storage Containers—Tight containers.Storage—Light-resistant.
Carmellose Calciumカルメロースカルシウム
Change the Description to read:
Description Carmellose Calcium occurs as a white toyellowish white powder.
It is practically insoluble in ethanol (95) and in diethylether.
It swells with water to form a suspension.The pH of a suspension, obtained by shaking 1.0 g of
Carmellose Calcium with 100 mL of water, is between 4.5and 6.0.
It is hygroscopic.
Change the Identiˆcation (1) and (4) to read:
Identiˆcation (1) Shake thoroughly 0.1 g of CarmelloseCalcium with 10 mL of water, followed by 2 mL of sodiumhydroxide TS, allow to stand for 10 minutes, and use thissolution as the sample solution. To 1 mL of the samplesolution add water to make 5 mL. To 1 drop of this solutionadd 0.5 mL of chromotropic acid TS, and heat in a waterbath for 10 minutes: a red-purple color develops.
(4) Ignite 1 g of Carmellose Calcium to ash, dissolve theresidue in 10 mL of water and 6 mL of acetic acid (31), andˆlter, if necessary. Boil the ˆltrate, cool, and neutralize withammonia TS: the solution responds to the Qualitative Tests(1) and (3) for calcium salt.
Change the Purity to read:
Purity (1) Alkali—Shake thoroughly 1.0 g of CarmelloseCalcium with 50 mL of freshly boiled and cooled water, andadd 2 drops of phenolphthalein TS: no red color develops.
(2) Chloride—Shake thoroughly 0.80 g of CarmelloseCalcium with 50 mL of water, add 10 mL of sodiumhydroxide TS to dissolved, add water to make 100 mL, anduse this solution as the sample solution. Heat 20 mL of thesample solution with 10 mL of 2 mol/L nitric acid on awater bath until a ‰occulent precipitate is produced. Aftercooling, centrifuge, and take out the supernatant liquid.Wash the precipitate with three 10-mL portions of water bycentrifuging each time, combine the supernatant and thewashings, and add water to make 100 mL. Take 25 mL ofthis solution, and add 1 mL of nitric acid and water to make50 mL. Perform the test using this solution as the testsolution. Prepare the control solution with 0.40 mL of 0.01mol/L hydrochloric acid VS (not more than 0.36z).
(3) Sulfate—Heat 10 mL of the sample solution ob-tained in (2) with 1 mL of hydrochloric acid in a water bathuntil a ‰occulent precipitate is produced. Cool, centrifuge,and take out the supernatant liquid. Wash the precipitatewith three 10-mL portions of water by centrifuging eachtime, combine the supernatant and the washings, and addwater to make 100 mL. Perform the test with 25 mL thissolution as the test solution. Prepare the control solutionwith 0.42 mL of 0.005 mol/L sulfuric acid VS. To the testsolution and the control solution add 1 mL of 3 mol/Lhydrochloric acid and 3 mL of barium chloride TS, then addwater to make 50 mL, and mix. Allow to stand for 10minutes, and compare the turbidity of these solutions: theturbidity obtained with the test solution is not more thanthat obtained with the control solution (not more than1.0z).
(4) Heavy metals—Proceed with 1.0 g of CarmelloseCalcium according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard Lead
17601760 Supplement II, JPXIVO‹cial Monographs for Part II
Solution (not more than 20 ppm).
Change the Residue on ignition to read:
Residue on ignition 10.0 – 20.0z (after drying, 1 g).
Cellulose Acetate Phthalate酢酸フタル酸セルロース
Add the following commonly used name:
Cellacefate
Change the origin/limits of the content to read:
Cellulose Acetate Phthalate is a reaction product ofphthalic anhydride and partially acetylated cellulose.
Cellulose Acetate Phthalate, calculated on theanhydrous and free acid-free basis, contains not lessthan 21.5z and not more than 26.0z of acetyl group(-COCH3: 43.04), and not less than 30.0z andnot more than 36.0z of carboxybenzoyl group(-COC6H4COOH: 149.12).
Change the Description to read:
Description Cellulose Acetate Phthalate occurs as a whitepowder or grain.
It is freely soluble in acetone, and practically insoluble inwater, in methanol and in ethanol (99.5).
Change the Identiˆcation to read:
Identiˆcation Determine the infrared absorption spectrumof Cellulose Acetate Phthalate as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor spectrum of Cellulose Acetate Phthalate ReferenceStandard: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
Change the Water to read:
Water Not more than 5.0z (1 g, volumetric titration,direct titration, using a mixture of ethanol (99.5) anddichloromethane (3:2) instead of methanol for Karl Fischermethod).
Change the Assay (2) to read:
Assay(2) Acetyl group—Weigh accurately about 0.1 g of
Cellulose Acetate Phthalate, put in a glass-stoppered conical‰ask, add exactly 25 mL of 0.1 mol/L sodium hydroxideVS, and boil for 30 minutes under a re‰ux condenser. Aftercooling, add 5 drops of phenolphthalein TS, and titrate with0.1 mol/L hydrochloric acid VS. Perform a blank determi-nation.
Content (z) of free acids and bound acetyl group (C2H3O)
＝AW×0.4304
A: Amount (mL) of 0.1 mol/L sodium hydroxide con-sumed, corrected by the blank determination
W: Amount (g) of the test sample, calculated on theanhydrous basis
Content (z) of acetyl group (C2H3O)
＝P－0.5182×B
100－B×100－0.5772×C
B: Amount (z) of free acids obtained in Purity (2) Freeacids
C: Content (z) of carboxybenzoyl groupP: Content (z) of free acids and bound acetyl group
(C2H3O)
Chrysanthemum Flowerキクカ
Change the Extract content to read:
Extract content Dilute ethanol-extract: not less than30.0z.
Add the following:
Cnidium Monnieri Fruit
Cnidii Monnieris Fructus
ジャショウシ
Cnidium Monnieri Fruit is the fruit of Cnidiummonnieri Cusson (Umbelliferae).
Description Elliptical cremocarp, often each mericarpseparated; 2 – 3 mm in length, 1 – 2 mm in width; externallylight brown to brown, each mericarp usually with ˆvewinged longitudinal ridges; inner surface of mericarp almost‰at.
Odor, characteristic; it gives characteristic aroma, later aslight sensation of numbness on chewing.
Under a microscope, a transverse section reveals one oilcanal between longitudinal ridges, usually two oil canals inthe inner part of mericarp facing to gynophore; longitudinalridges composed of slightly ligniˆed parenchymatous cells,with vascular bundles in the base; epidermal cells andparenchymatous cells of longitudinal ridges contain solitarycrystals of calcium oxalate; parenchymatous cells ofalbumen contain oil drops and aleurone grains, and oc-casionally starch grains.
Identiˆcation To 1 g of pulverized Cnidium MonnieriFruit add 10 mL of ethyl acetate, shake for 10 minutes,ˆlter, and use the ˆltrate as the sample solution. Separately,
17611761Supplement II, JPXIV O‹cial Monographs for Part II
dissolve 1 mg of osthole for thin-layer chromatography in2 mL of methanol, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 5 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography, develop the platewith a mixture of hexane and ethyl acetate (2:1) to a distanceof about 10 cm, and air-dry the plate. Examine under ultrav-iolet light (main wavelength: 365 nm): one of the spot amongthe several spots from the sample solution has the same colortone and the Rf value with the blue-white ‰uorescent spotfrom the standard solution.
Loss on drying Not more than 12.0z (6 hours).
Total ash Not more than 17.0z.
Acid-insoluble ash Not more than 6.0z.
Extract content Not less than 8.0z (dilute ethanol-solubleextract).
Change to read:
Corn Starch
Amylum Maydis
トウモロコシデンプン
Corn Starch consists of starch granules derived fromthe ripen seeds of Zea mays Linn áe (Gramineae).
Description Corn Starch occurs as white to pale yellowishwhite masses or powder.
It is practically insoluble in water and in ethanol (99.5).
Identiˆcation (1) Under a microscope, Corn Starch,preserved in a mixture of water and glycerin (1:1), appears asirregularly polygonal simple grains usually 2 – 23 mm indiameter, or irregularly orbicular or spherical simple grains25 – 35 mm in diameter; hilum appears as distinct cave or 2 –5 radial clefts; concentric striation absent; a black cross, itsintersection point on hilum, is observed when grains are putbetween two polarizing prisms ˆxed at right angle to eachother.
(2) To 1 g of Corn Starch add 50 mL of water, boil for 1minute, and allow to cool: a subtle white-turbid, pasty liquidis formed.
(3) To 10 mL of the pasty liquid obtained in (2) add0.04 mL of iodine TS: a orange-red to dark blue-purplecolor is formed, and the color disappears by heating.
pH Put 5.0 g of Corn Starch in a non-metal vessel, add25.0 mL of freshly boiled and cooled water, mix gently for 1minute, and allow to stand for 15 minutes: the pH of thesolution is between 4.0 and 7.0.
Purity (1) Iron—To 1.5 g of Corn Starch add 15 mL of 2mol/L hydrochloric acid TS, mix, ˆlter, and use the ˆltrateas the test solution. To 2.0 mL of Standard Iron Solution
add water to make 20 mL, and use as the control solution.Put 10 mL each of the test solution and the control solutionin test tubes, add 2 mL of a solution of citric acid (2 in 10)and 0.1 mL of mercapto acetic acid, and mix. Alkalize withammonia solution (28) to litmus paper, add water to make20 mL, and mix. Transfer 10 mL each of these solutions intotest tubes, allow to stand for 5 minutes, and compare thecolor of these solutions against a white background: thecolor of the test solution is not darker than that of thecontrol solution (not more than 10 ppm).
(2) Oxidizing substances—To 4.0 g of Corn Starch add50.0 mL of water, shake for 5 minutes, and centrifuge. To30.0 mL of the supernatant liquid add 1 mL of acetic acid(100) and 0.5 to 1.0 g of potassium iodide, shake, and allowto stand for 25 to 30 minutes at a dark place. Add 1 mL ofstarch TS, and titrate with 0.002 mol/L sodium thiosulfateVS until the color of the solution disappears. Perform ablank determination and make any necessary correction: thevolume of 0.002 mol/L sodium thiosulfate VS consumed isnot more than 1.4 mL (not more than 20 ppm, calculated ashydrogen peroxide).
(3) Sulfur dioxide—(i) Apparatus Use as shown in the following ˆgure.
(ii) Procedure Introduce 150 mL of water into theboiling ‰ask, close the tap of the funnel, and pass carbondioxide through the whole system at a rate of 100±5 mL perminute. Pass cooling water through the condenser, and place10 mL of hydrogen peroxide-sodium hydroxide TS in thetest-tube. After 15 minutes, remove the funnel withoutinterrupting the stream of carbon dioxide, and introducethrough the opening into the ‰ask about 25 g of CornStarch, accurately weighed, with the aid of 100 mL of water.Apply tap grease to the outside of the connection part of the
17621762 Supplement II, JPXIVO‹cial Monographs for Part II
funnel, and load the funnel. Close the tap of the funnel,pour 80 mL of 2 mol/L hydrochloric acid TS into thefunnel, open the tap to introduce the hydrochloric acid intothe ‰ask, and close the tap while several mL of thehydrochloric acid remains, in order to avoid losing sulfurdioxide. Place the ‰ask in a water bath, and heat the mixturefor 1 hour. Transfer the contents of the test-tube with the aidof a little water to a wide-necked conical ‰ask. Heat in awater bath for 15 minutes, and cool. Add 0.1 mL ofbromophenol blue TS, and titrate with 0.1 mol/L sodiumhydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determi-nation and make any necessary correction. Calculate theamount of sulfur dioxide by applying the following formula:it is not more than 50 ppm.
Amount (ppm) of sulfur dioxide＝VW×1000×3.203
W: Amount (g) of the sampleV: Amount (mL) of 0.1 mol/L sodium hydroxide VS
consumed
Loss on drying Not more than 15.0z (1 g, 1309C, 90minutes).
Reisdue on ignition Not more than 0.6z (1 g).
Containers and storage Containers—Well-closed contain-ers.
Cornus Fruitサンシュユ
Change the Identiˆcation to read:
Identiˆcation To 1 g of coarse cuttings of Cornus Fruitadd 10 mL of methanol, shake for 5 minutes, ˆlter, and usethe ˆltrate as the sample solution. Separately, dissolve 1 mgof loganin for thin-layer chromatography in 2 mL ofmethanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop with a mixture of ethylacetate, water and formic acid (6:1:1) to a distance of about10 cm, and air-dry the plate. Spray evenly 4-methoxybenzal-dehyde-sulfuric acid TS on the plate, and heat at 1059C for 5minutes: one of the spots from the sample solution is thesame with a red-purple spot from the standard solution incolor tone and Rf value.
Delete the following Monographs:
Digitalis
Powdered Digitalis
Add the following:
Epimedium Herb
Epimedii Herba
インヨウカク
Epimedium Herb is the terrestrial part of Epimedi-um pubescens Maximowicz, Epimedium brevicornumMaximowicz, Epimedium wushanense T. S. Ying,Epimedium sagittatum Maximowicz, Epimediumkoreanum Nakai, Epimedium grandi‰orum Morrenvar. thunbergianum Nakai or Epimedium sempervi-rens Nakai (Berberidaceae).
Description Epimedium Herb is composed of a stem and aternate to triternate compound leaf; lea‰et ovate to broadlyovate or ovate-lanceolate, 3 – 20 cm in length, 2 – 8 cm inwidth, petiolule 15 – 70 mm in length, apex of lea‰etacuminate, needle hair on margin 0.1 – 0.2 cm in length,base of lea‰et cordate to deeply cordate, lateral lea‰et asym-metry; upper surface green to greenish brown, sometimeslustrous, lower surface light green, often pilose, especiallyon vein densely pilose, papery or coriaceous; petiole andstem cylindrical, light yellowish brown to slightly purplishand light greenish brown, easily broken.
Odor, slight; taste, slightly bitter.Under a microscope, a transverse section of the leaf
reveals 3 – 6 vascular bundles in midvein; mesophyllcomposed of upper epidermis, single-layered palisade,spongy tissue and lower epidermis; leaf margins orbicular oroblong, sclerenchymatous; multi-cellular hairs on epidermis;8 – 20 vascular bundles in petiole and 6 – 15 vascular bundlesin petiolule. Under a microscope, a transverse section of thestem reveals a single to several-layered hypodermis, cortexof 4 – 10 layers of sclerenchymatous cells, vascular bundle13 – 30 in number, oblong to obovate.
Identiˆcation To 2 g of pulverized Epimedium Herb add20 mL of methanol, shake for 15 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 1 mg oficariin for thin-layer chromatography in 1 mL of methanol,and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of silica gel with ‰uores-cent indicator for thin-layer chromatography. Develop theplate with a mixture of ethyl acetate, ethanol (99.5) andwater (8:2:1) to a distance of about 10 cm, and air-dry theplate. Examine under ultraviolet light (main wavelength: 254
17631763Supplement II, JPXIV O‹cial Monographs for Part II
nm): one spot among the spots from the sample solution anda dark purple spot from the standard solution show the samecolor tone and the same Rf value.
Loss on drying Not more than 12.5z (6 hours).
Total ash Not more than 8.5z.
Acid-insoluble ash Not more than 2.0z.
Extract content Not less than 17.0z (dilute ethanol-solu-ble extract).
Powdered Gardenia Fruitサンシシ末
Change the origin/limits of content to read:
Powdered Gardenia Fruit is the powder of GardeniaFruit.
It contains not less than 3.0z of geniposide, calcu-lated on the basis of dried material.
Change the Identiˆcation (1) to read:
Identiˆcation (1) To 1.0 g of Powdered Gardenia Fruit,previously dried in a desiccator (silica gel) for 24 hours, add100 mL of hot water, warm the mixture between 609C and709C for 30 minutes with frequent shaking, and ˆlter aftercooling. To 1.0 mL of the ˆltrate add water to make 10 mL:the color of the resulting solution is yellow and is not lighterthan that of the following control solution.
Control solution: Dissolve 9.8 mg of carbazochromesodium sulfonate in water to make exactly 10 mL. Pipet 1mL of this solution, and add water to make exactly 50 mL.
Add the following next to Identiˆcation:
Loss on drying Not more than 13.0z.
Add the following next to Total ash:
Component determination Weigh accurately about 0.5 gof Powdered Gardenia Fruit, transfer into a glass-stopperedcentrifuge tube, add 40 mL of diluted methanol (1 in 2),shake for 15 minutes, centrifuge, and take the supernatantliquid. To the residue add 40 mL of diluted methanol (1 in2), and perform as the same as above. Combine the extractsso obtained, and add diluted methanol (1 in 2) to makeexactly 100 mL. Pipet 5 mL of the solution, add methanol tomake exactly 20 mL, use this solution as the samplesolution. Separately, weigh accurately about 10 mg ofgeniposide for component determination, previously driedin a desiccator (in vacuum, phosphorus (V) oxide) for 24hours, and dissolve in methanol to make exactly 100 mL.Pipet 5 mL of the solution, add methanol to make exactly 10mL, and use this solution as the standard solution. Performthe test with exactly 10 mL each of the sample solution andthe standard solution as directed under the Liquid
Chromatography according to the following conditions, andmeasure the peak areas of geniposide, AT and AS, of bothsolutions.
Amount (mg) of geniposide＝WS×AT
AS×2
WS: Amount (mg) of geniposide for component determi-nation
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 240 nm).Column: A stainless steel column 6 mm in inside diameter
and 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about309C.
Mobile phase: A mixture of water and acetonitrile (22:3).Flow rate: Adjust the ‰ow rate so that the retention time
of geniposide is about 15 minutes.System suitability—
System performance: Dissolve 1 mg each of geniposidefor component determination and caŠeine in methanol tomake 15 mL. When the procedure is run with 10 mL of thissolution under the above operating conditions, caŠeine andgeniposide are eluted in this order with the resolutionbetween these peaks being not less than 3.5.
System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of geniposide is not more than 1.5z.
Gingerショウキョウ
Change the Identiˆcation to read:
Identiˆcation To 2 g of pulverized Ginger add 5 mL ofdiethyl ether, shake for 10 minutes, ˆlter, and use the ˆltrateas the sample solution. Separately, dissolve 1 mg of [6]-gin-gerol for thin-layer chromatography in 2 mL of methanol,and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL of the sample solution and thestandard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethylacetate and hexane (1:1) to a distance of about 10 cm, andair-dry the plate. Spray evenly 4-dimethylaminobenzalde-hyde TS on the plate, heat at 1059C for 5 minutes, and allowto cool: one of the spots from the sample solution and agreen spot from the standard solution show the same colortone and Rf value.
17641764 Supplement II, JPXIVO‹cial Monographs for Part II
Powdered Gingerショウキョウ末
Change the Identiˆcation to read:
Identiˆcation To 2 g of Powdered Ginger add 5 mL ofdiethyl ether, shake for 10 minutes, ˆlter, and use the ˆltrateas the sample solution. Separately, dissolve 1 mg of [6]-gin-gerol for thin-layer chromatography in 2 mL of methanol,and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL of the sample solution and thestandard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethylacetate and hexane (1:1) to a distance of about 10 cm, andair-dry the plate. Spray evenly 4-dimethylaminobenzalde-hyde TS on the plate, heat at 1059C for 5 minutes, and allowto cool: one of the spots from the sample solution and agreen spot from the standard solution show the same colortone and Rf value.
Chorionic Gonadotrophin胎盤性性腺刺激ホルモン
Delete the origin/limits of content and add thefollowing:
Method of preparation Chorionic Gonadotrophin is adried preparation of gonad-stimulating hormone obtainedfrom the urine of healthy pregnant women after themanufacturing process to remove or inactivate the virus. Itcontains not less than 2500 chorionic gonadotrophin Unitsper mg, and contains not less than 3000 chorionicgonadotrophin Units per mg protein.
It contains not less than 80z and not more than 125z ofthe labeled Units of chorionic gonadotrophin.
Change the Description to read:
Description Chorionic Gonadotrophin occurs as a white tolight yellow-brown powder.
It is freely soluble in water.
Delete the Toxicity and Pyrogen, and add thefollowing:
Bacterial endotoxins Less than 0.03 EU/unit.
Abnormal toxicity Dilute Chorionic Gonadotrophin withisotonic sodium chloride solution so that each mL of thesolution contains 120 Units, and use this solution as thesample solution. Inject 5.0 mL of the sample solution intothe peritoneal cavity of each of 2 or more of well-nourished,healthy guinea pigs weighing about 350 g, and observe theconditions of the animals for more than 7 days: all theanimals exhibit no abnormalities.
Speciˆc activity When calculate from the results obtainedby the Assay and the following test, the speciˆc activity isnot less than 3000 chorionic gonadotrophin Units per mgprotein.
(i) Sample solution—To an exactly amount of ChorionicGonadotrophin add water to make a solution so that eachmL contains about 500 Units of chorionic gonadotrophinaccording to the labeled amount.
(ii) Standard solution—Weigh accurately about 10 mgof bovine serum albumin, and dissolve in water to makeexactly 20 mL. To a suitable volume of this solution addwater to make four solutions containing exactly 300, 200,100 and 50 mg of the albumin per mL, respectively.
(iii) Procedure—Pipet 0.5 mL each of the sample solu-tion and the standard solutions, put them in glass test tubesabout 18 mm in inside diameter and about 130 mm in length,add exactly 5 mL of alkaline copper TS, mix, and allow thetubes to stand in a water bath at 309C for 10 minutes. Thenadd exactly 0.5 mL of diluted Folin's TS (1 in 2), mix, andwarm in a water bath at 309C for 20 minutes. Determine theabsorbances of these solutions at 750 nm as directed underthe Ultraviolet-visible Spectrophotometry using a solutionobtained in the same manner with 0.5 mL of water as theblank.
Plot the absorbances of the standard solutions on thevertical axis and their protein concentrations on the horizon-tal axis to prepare a calibration curve, and determine theprotein content of the sample solution from its absorbanceby using this curve. Then calculate the amount of the proteinin the sample.
Change the Assay (i) to read:
Assay (i) Test animals—Select healthy female albino ratsweighing about 45 to 65 g.
Description Chorionic Gonadotrophin for Injectionoccurs as a white to light yellow-brown powder or masses.
Add the following next to Identiˆcation:
pH Prepare a solution so that each mL of isotonic sodiumchloride solution contains 2 mg of Chorionic Gonadotoro-phin for Injection: the pH of this solution is between 5.0 and7.0.
Delete the Pyrogaen and add the following:
Bacterial endotoxins Less than 0.03 EU/unit.
Mass variation When calculate the acceptance value usingthe mean of estimated contents of the units tested as M, it
17651765Supplement II, JPXIV O‹cial Monographs for Part II
meets the requirements of the Mass Variation Test.
Foreign insoluble matter Perform the test according toMethod 2: it meets the requirements of the Foreign InsolubleMatter Test for Injections.
Insoluble particulate matter Perform the test according toMethod 1: it meets the requirements of the Insoluble Par-ticulate Matter Test for Injections.
Sterility Perform the test according to the Membraneˆltration method: it meets the requirements of the SterilityTest.
Serum Gonadotrophin血清性性腺刺激ホルモン
Change the origin/limits of content to read:
Method of preparation Serum Gonadotrophin is a driedpreparation of gonad-stimulating hormone, obtained frompregnant mares' serum which has adequately inspectedviruses, and subjected to a suitable process for removal orinactivation of viruses. It contains not less than 2000 serumgonadotrophin Units per mg.
It contains not less than 80z and not more than 125z ofthe labeled Units of serum gonadotrophin.
Change the Description to read:
Description Serum Gonadotrophin occurs as a whitepowder.
It is freely soluble in water.
Add the following next to Purity:
Speciˆc activity When calculated from the results obtainedby the Assay and the following test, Serum Gonadotrophincontaines not less than 3000 serum gonadotrophin Units permg of protein.
(1) Standard solutions—Dissolve about 3 mg of bovineserum albumin in water to make a solution containing500 mg of the albumin in each mL. To this solution addwater to make four standard solutions so that each mLcontains exactly 200 mg, 150 mg, 100 mg and 50 mg of thealbumin, respectively.
(2) Sample solution—Dissolve about 1 mg of SerumGonadotrophin in water to make a solution containingexactly 180 mg in each mL.
(3) Sodium carbonate solution—Dissolve 2 g of sodiumcarbonate (standard reagent) in 0.1 mol/L sodiumhydroxide TS to make 100 mL.
(4) Sodium tartrate solution—Dissolve about 1 g ofsodium tartrate dihydrate in water to make 100 mL.
(5) Copper (II) sulfate solution—Dissolve 0.5 g ofcopper (II) sulfate pentahydrate in the sodium tartratesolution to make 100 mL.
(6) Alkaline copper solution—Mix 50 mL of the sodiumcarbonate solution and 1 mL of the copper (II) sulfate
solution. Prepare before use. Use within the day of prepara-tion.
(7) Procedure—Pipet 0.5 mL each of the standardsolutions and the sample solution in small test tubes, add3 mL of the alkaline copper solution to them, and mix.Allow them to stand at the room temperature for not lessthan 10 minutes, add 0.3 mL of diluted Folin's TS (1 in 2),mix immediately, and allow to stand for not less than 30minutes. Determine the absorbances of these solutions soobtained at 750 nm as directed under the Ultraviolet-visibleSpectrophotometry using a solution, prepared in the samemanner with 0.5 mL of water, as the blank. Plot the calibra-tion curve from the absorbances obtained with the standardsolutions, and determine the amount of protein in thesample solution from this curve.
Speciˆc activity (unit/mg protein)
＝units per mg, obtained in the Assayamount (z) of protein in the sample
×100
Change the Toxicity to read:
Abnormal toxicity Dissolve Serum Gonadotrophin inisotonic sodium chloride solution so that each 5 mL of thesolution contains 4000 Units, and use this solution as thesample solution. Inject 5.0 mL of the sample solution intothe peritoneal cavity of each of 2 or more of well-nourished,healthy guinea pigs weighing about 350 g, and inject 0.5 mLof the sample solution into the peritoneal cavity of each of 2or more of well-nourished, healthy mice aged about 5 weeks.Observe the conditions of the animals for more than 7 days:all the animals exhibit no abnormalities.
Change the Pyrogen to read:
Bacterial endotoxins Less than 0.1 EU/unit.
Serum Gonadotrophin for Injection注射用血清性性腺刺激ホルモン
Change the Description to read:
Description Serum Gonadotrophin for Injection occurs aswhite powder or masses.
Add the following next to Identiˆcation:
pH Dissolve 30 mg of Serum Gonadotrophin for Injectionin 20 mL of isotonic sodium chloride solution: the pH of thissolution is between 5.0 and 7.0.
Delete the Pyrogen and add the following:
Bacterial endotoxins Less than 0.1 EU/unit.
17661766 Supplement II, JPXIVO‹cial Monographs for Part II
Add the following:
Lindera Root
Linderae Radix
ウヤク
Lindera Root is the root of Lindera strychnifoliaFernandez-Villar (Lauraceae).
Description Fusiform or rosary-like root, 10 – 15 cm inlength, 10 – 25 mm in diameter; externally yellowish brownto brown, with a few scars of rootlets; a transverse sectionreveals cortex brown, xylem light yellowish brown, concen-tric circles and radially arranged lines brown; dense and hardin texture.
Odor, camphor-like; taste, bitter.Under a microscope, a transverse section of the root with
periderm reveals a cork layer several cells thick, partiallyconsisting of cork stone cells; cortex parenchyma sometimescontains oil cells and ˆbers; in xylem, vessels-xylem ˆbersand rays are arranged alternately; parenchyatous cells ofcortex and xylem contain sandy and columnar crystals ofcalcium oxalate, simple starch grains 1 – 15 mm in diameter,and 2- to 4- compound starch grains.
Identiˆcation To 3 g of pulverized Lindera Root add40 mL of hexane, and warm under a re‰ux condenser on awater bath for 30 minutes. After cooling, ˆlter, to theresidue add 10 mL of ammonia TS and 30 mL of a mixtureof ethyl acetate and diethyl ether (1:1), shake vigorously for20 minutes, and centrifuge. Separate the supernatant liquid,add 10 g of anhydrous sodium sulfate, shake, and ˆlter.Evaporate the ˆltrate, dissolve the residue with 0.5 mL ofethanol (99.5), and use this solution as the sample solution.Perform the test with this solution as directed under theThin-layer Chromatography. Spot 20 mL of the samplesolution on a plate of silica gel for thin-layer chro-matography, develop the plate with a mixture of ethylacetate, methanol and ammonia water (28) (10:2:1) to adistance of about 10 cm, and air-dry the plate. Spray evenlyDragendorŠ's TS for spraying on the plate: a yellow-brownspot appears at around Rf 0.4.
Loss on drying Not more than 14.0z (6 hours).
Total ash Not more than 2.5z.
Extract content Not less than 6.0z (dilute ethanol-solubleextract).
Add the following:
Lonicera Leaf and Stem
Lonicerae Folium Cum Caulis
ニンドウ
Lonicera Leaf and Stem is the leaves and stems ofLonicera japonica Thunberg (Caprifoliaceae).
Description Leaves and opposite leaves on short stem;leaf, ovate and entire, with short petiole, 3 – 7 cm in length,1 – 3 cm in width; upper surface greenish brown, lowersurface light grayish green; under a magnifying glass, bothsurfaces pubescent. Stem, 1 – 4 mm in diameter; externallygrayish yellow-brown to purplish brown, a transversesection of stem, round and hollow.
Almost odorless; taste, slightly astringent, followed by alitter bitterness.
Under a microscope, a transverse section of leaf revealsthe outermost layer of upper and lower surfaces to be com-posed of a single-layered epidermis, uni-cellular non-glandu-lar hairs and multi-cellular glandular hairs on epidermis; inmidvein, several-layered collenchyma present beneath theepidermis and vascular bundles in the center; in mesophyll,palisade layer adjacent to upper epidermis, spongy tissue ad-jacent to lower epidermis; glandular hairs contain brownsecretion, parenchymatous cells contain aggregate crystalsof calcium oxalate, and occasionally starch grains.
Identiˆcation To 1 g of pulverized Lonicera Leaf and Stemadd 5 mL of methanol, shake for 5 minutes, centrifuge, anduse the supernatant liquid as the sample solution. Separate-ly, dissolve 1 mg of chlorogenic acid for thin-layer chro-matography in 2 mL of methanol, and use this solution asthe standard solution (1). Separately, dissolve 1 mg ofloganin for thin-layer chromatography in 2 mL of methanol,and use this solution as the standard solution (2). Performthe test with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solution,the standard solution (1) and the standard solution (2) on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of ethyl acetate, water and formicacid (6:1:1) to a distance of about 10 cm, and air-dry theplate. Examine under ultraviolet light (main wavelength:365 nm): one of the spot among the several spots from thesample solution has the same color tone and Rf value withthe blue-white ‰uorescent spot from the standard solution(1). Spray evenly 4-methoxybenzaldehyde-sulfuric acid TSon the plate, and heat at 1059C for 5 minutes: one of thespot among the several spots from the sample solution hasthe same color tone and Rf value with the red-purple spotfrom the standard solution (2).
Purity Stem—Lonicera Leaf and Stem does not containsthe stems larger than 5 mm in diameter.
Loss on drying Not more than 12.0z (6 hours).
17671767Supplement II, JPXIV O‹cial Monographs for Part II
Total ash Not more than 9.0z.
Acid-insoluble ash Not more than 1.0z.
Extract content Not less than 12.0z (dilute ethanol-solu-ble extract).
Add the following:
Lycium Bark
Lycii Cortex
ジコッピ
Lycium Bark is the root bark of Lycium chinenseMiller or Lycium barbarum Linn áe (Solanaceae).
Description Tubular to semitubular bark, 1 – 6 mm inthickness; externally light brown to light yellowish brown,periderm peeled easily as scale; internally grayish brown,longitudinally striate; brittle in texture; fractured surface,grayish white, not ˆbrous.
Odor, weak and characteristic; taste, slightly sweet at ˆrst.Under a microscope, a transverse section reveals periderm
composed of a cork layer of several layers of thin walledcork cells; in cortex parenchymatous cells containing sandycrystals of calcium oxalate sparsely distributed, occasionallya few ˆbers observed; parenchymatous cells contain starchgrains, 1 – 10 mm in diameter; stone cells very rare.
Identiˆcation To 1.0 g of pulverized Lycium Bark add 10mL of methanol, shake for 15 minutes, ˆlter, and use theˆltrate as the sample solution. Perform the test with thissolution as directed under the Thin-layer Chromatography.Spot 10 mL of the sample solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of 1-butanol, water, pyridine and acetic acid (100)(3:1:1:1) to a distance of about 10 cm, and air-dry the plate.Spray evenly DragendorŠ's TS for spraying on the plate,heat at 1059C for 3 minutes, then spray evenly sodium nitriteTS: a dark brown principal spot appears at around Rf 0.5.
Loss on drying Not more than 11.5z (6 hours).
Total ash Not more than 20.0z.
Acid-insoluble ash Not more than 3.0z.
Extract content Not less than 10.0z (dilute ethanol-solu-ble extract).
Add the following:
Lycium Fruit
Lycii Fructus
クコシ
Lycium Fruit is the fruit of Lycium chinense Milleror Lycium barbarum Linne (Solanaceae).
Description Fusiform fruit with acute apex, 6 – 20 mm inlength, 3 – 8 mm in diameter, pericarp red to dark red,externally roughly wrinkled; under a magnifying glass, atransverse section of fruit reveals two locules containingnumerous seeds; seed light brown to light yellowish brown,about 2 mm in a diameter, compressed reniform.
Odor, characteristic; taste, sweet, later slightly bitter.
Identiˆcation To 1.0 g of powdered Lycium Fruit add 5mL of ethyl acetate, shake for 15 minutes, ˆlter, and use theˆltrate as the sample solution. Perform the test with thissolution as directed under the Thin-layer Chromatography.Spot 20 mL of the sample solution on a plate of silica gel forthin-layer chromatography, develop the plate with a mixtureof hexane and ethyl acetate (10:1) to a distance of about10 cm, and air-dry the plate: a yellow principal spot appearsat around Rf 0.6.
Purity Foreign matter—It contains not more than 2.0z offoreign matter such as peduncle or others.
Total ash Not more than 8.0z.
Acid-insoluble ash Not more than 1.0z.
Extract content Not less than 35.0z (dilute ethanol-solu-ble extract).
Opium Alkaloids Hydrochlorides塩酸アヘンアルカロイド
Change the origin/limits of content to read:
Opium Alkaloids Hydrochlorides consists of thehydrochlorides of some of the main alkaloids obtainedfrom opium.
It contains not less than 47.0z and not more than52.0z of morphine (C17H19NO3: 285.34), and not lessthan 35.0z and not more than 41.0z of other opiumalkaloids.
Change the Description to read:
Description Opium Alkaloids Hydrochlorides occur as awhite to light brown powder.
It is soluble in water, and slightly soluble in ethanol (99.5).It is colored by light.
17681768 Supplement II, JPXIVO‹cial Monographs for Part II
Change the Identiˆcation (1) to read:
Identiˆcation (1) Dissolve 0.1 g of Opium AlkaloidsHydrochlorides in 10 mL of diluted ethanol (1 in 2), and usethis solution as the sample solution. Separately, dissolve60 mg of Morphine Hydrochloride, 40 mg of NoscapineHydrochloride, 10 mg of Codein Phosphate and 10 mg ofPapaverine Hydrochloride in 10 mL each of diluted ethanol(1 in 2), and use these solutions as the standard solutions (1),(2), (3) and (4), respectively. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 20 mL each of the sample solution and the standardsolutions on a plate of silica gel with ‰uorescent indicatorfor thin-layer chromatography. Develop the plate with amixture of acetone, toluene, ethanol (99.5) and ammoniasolution (28) (20:20:3:1) to a distance of about 10 cm, andair-dry the plate. Examine under ultraviolet light (mainwavelength: 254 nm): each spot from the sample solution isthe same in color tone and Rf value with the correspondingspot from the standard solutions (1), (2), (3) and (4) (mor-phine, noscapine, codeine and papaverine).
Change the Assay to read:
Assay Weigh accurately about 0.1 g of Opium AlkaloidsHydrochlorides, and dissolve in water to make exactly 50mL, and use this solution as the sample solution. Separately,weigh accurately about 60 mg of morphine hydrochloridefor assay, dissolve in water to make exactly 50 mL, and usethis solution as the standard solution. Perform the test withexactly 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine the peakareas of morphine, codeine, papaverine, thebaine, narceineand noscapine, AT1, AT2, AT3, AT4, AT5 and AT6, from thesample solution, and the peak area of morphine, AS, fromthe standard solution.
Amount (mg) of morphine (C17H19NO3)
＝WS×AT1
AS×0.887
Amount (mg) of other opium alkaloids＝WS
×AT2＋AT3×0.29＋AT4×0.20＋AT5×0.19＋AT6
AS×0.887
WS: Amount (mg) of morphine hydrochloride for assay,calculated on the anhydrous basis
The relative retention time of codine, papaverine, the-baine, narceine and noscapine with respect to morphineobtained under the following operating conditions are asfollows.
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 285 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in500 mL of diluted phosphoric acid (1 in 1000), and adjustthe pH to 3.0 with sodium hydroxide TS. To 240 mL of thissolution add 70 mL of tetrahydrofuran, and mix.
Flow rate: Adjust the ‰ow rate so that the retention timeof morphine is about 10 minutes.System suitability—
System performance: Dissolve 60 mg of MorphineHydrochloride, 10 mg of Codeine Phosphate, 10 mg ofPapaverine Hydrochloride and 40 mg of NoscapineHydrochloride in water to make 50 mL. When the procedureis run with 20 mL of this solution under the above operatingconditions, morphine, codeine, papaverine and noscapineare eluted in this order with the complete separation betweenthese peaks and with the resolution between the peaks ofmorphine and codeine being not less than 1.5.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of morphine is not more than 1.0z.
Powdered Peony Rootシャクヤク末
Change the origin/limits of content to read:
Powdered Peony Root is the powder of Peony Root.It contains not less than 2.0z of paeoni‰orin
(C23H28O11: 480.46), calculated on the dried basis.
Change the Assay to read:
Assay Weigh accurately about 0.5 g of Powdered PeonyRoot, add 50 mL of diluted methanol (1 in 2), heat under are‰ux condenser on a water bath for 30 minutes, cool, andˆlter. To the residue add 50 mL of diluted methanol (1 in 2),and proceed in the same manner. Combine the ˆltrates, adddiluted methanol (1 in 2) to make exactly 100 mL, and usethis solution as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoni‰orin Reference Standard(separately detarmine the water content), dissolve in dilutedmethanol (1 in 2) to make exactly 100 mL, and use thissolution as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography
17691769Supplement II, JPXIV O‹cial Monographs for Part II
according to the following conditions. Determine the peakareas, AT and AS, of paeoni‰orin in each solution.
Amount (mg) of paeoni‰orin (C23H28O11)
＝WS×AT
AS
WS: Amount (mg) of Paeoni‰orin Reference Standard,calculated on the anhydrous basis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 232 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about209C.
Mobile phase: A mixture of water, acetonitrile andphosphoric acid (850:150:1).
Flow rate: Adjust the ‰ow rate so that the retention timeof paeoni‰orin is about 10 minutes.System suitability—
System performance: Dissolve 1 mg each of Paeoni‰orinReference Standard and albi‰orin in diluted methanol (1 in2) to make 10 mL. When the procedure is run with 10 mL ofthis solution under the above operating conditions, albi‰orinand paeoni‰orin are eluted in this order with the resolutionbetween these peaks being not less than 2.5.
System repeatability: When the test is repeated 6 timeswith the standard solution under the above operatingconditions, the relative standard deviation of the peak areaof paeoni‰orin is not more than 1.5z.
Change to read:
Potato Starch
Amylum Solani
バレイショデンプン
Potato Starch consists of starch granules derivedfrom the tuber of Solanum tuberosum Linn áe(Solanaceae).
Description Potato Starch occurs as a white powder.It is practically insoluble in water and in ethanol (99.5).
Identiˆcation (1) Under a microscope, Potato Starch,preserved in a mixture of water and glycerin (1:1), appears asunevenly ovoid or pyriform simple grains usually 30 – 100mm, often more than 100 mm in diameter, or spherical simplegrains 10 – 35 mm in diameter, rarely 2- to 4-compoundgrains; ovoid or pyriform simple grains with eccentric hilum,spherical simple grains with non-centric or slightly eccentrichilum; striation distinct in all grains; a black cross, itsintersection point on hilum, is observed when grains are put
between two polarizing prisms ˆxed at right angle to eachother.
(2) To 1 g of Potato Starch add 50 mL of water, boil for1 minute, and allow to cool: a subtle white-turbid, pastyliquid is formed.
(3) To 1 mL of the pasty liquid obtained in (2) add 0.05mL of iodine TS: a orange-red to dark blue-purple color isformed, and the color disappears by heating.
pH Put 5.0 g of Potato Starch in a non-metal vessel, add25.0 mL of freshly boiled and cooled water, mix gently for 1minute, and allow to stand for 15 minutes: the pH of thesolution is between 5.0 and 8.0.
Purity (1) Iron—To 1.5 g of Potato Starch add 15 mL of2 mol/L hydrochloric acid TS, mix, ˆlter, and use the ˆltrateas the test solution. To 2.0 mL of Standard Iron Solutionadd water to make 20 mL, and use as the control solution.Put 10 mL each of the test solution and the control solutionin test tubes, add 2 mL of a solution of citric acid (2 in 10)and 0.1 mL of mercapto acetic acid, and mix. Alkalize withammonia solution (28) to litmus paper, add water to make20 mL, and mix. Transfer 10 mL each of these solutions intotest tubes, allow to stand for 5 minutes, and compare thecolor of these solutions against a white background: thecolor of the test solution is not darker than that of thecontrol solution (not more than 10 ppm).
(2) Oxidizing substances—To 4.0 g of Potato Starch add50.0 mL of water, shake for 5 minutes, and centrifuge. To30.0 mL of the supernatant liquid add 1 mL of acetic acid(100) and 0.5 to 1.0 g of potassium iodide, shake, and allowto stand for 25 to 30 minutes at a dark place. Add 1 mL ofstarch TS, and titrate with 0.002 mol/L sodium thiosulfateVS until the color of the solution disappears. Perform ablank determination and make any necessary correction: thevolume of 0.002 mol/L sodium thiosulfate VS consumed isnot more than 1.4 mL (not more than 20 ppm, calculated ashydrogen peroxide).
(3) Sulfur dioxide—(i) Apparatus Use as shown in the following ˆgure.
17701770 Supplement II, JPXIVO‹cial Monographs for Part II
(ii) Procedure Introduce 150 mL of water into theboiling ‰ask, close the tap of the funnel, and pass carbondioxide through the whole system at a rate of 100±5 mL perminute. Pass cooling water through the condenser, and place10 mL of hydrogen peroxide-sodium hydroxide TS in thetest-tube. After 15 minutes, remove the funnel without inter-rupting the stream of carbon dioxide, and introduce throughthe opening into the ‰ask about 25 g of Potato Starch,accurately weighed, with the aid of 100 mL of water. Applytap grease to the outside of the connection part of thefunnel, and load the funnel. Close the tap of the funnel,pour 80 mL of 2 mol/L hydrochloric acid TS into thefunnel, open the tap to introduce the hydrochloric acid intothe ‰ask, and close the tap while several mL of thehydrochloric acid remains, in order to avoid losing sulfurdioxide. Place the ‰ask in a water bath, and heat the mixturefor 1 hour. Transfer the contents of the test-tube with the aidof a little water to a wide-necked conical ‰ask. Heat in awater bath for 15 minutes, and cool. Add 0.1 mL ofbromophenol blue TS, and titrate with 0.1 mol/L sodiumhydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determi-nation and make any necessary correction. Calculate theamount of sulfur dioxide by applying the following formula:it is not more than 50 ppm.
Amount (ppm) of sulfur dioxide＝VW×1000×3.203
W: Amount (g) of the sampleV: Amount (mL) of 0.1 mol/L sodium hydroxide VS
consumed
Loss on drying Not more than 20.0z (1 g, 1309C, 90
minutes).
Residue on ignition Not more than 0.6z (1 g).
Containers and storage Containers—Well-closed contain-ers.
Add the following:
Processed Aconite Root
Processi Aconiti Radix
ブシ
Processed Aconite Root is the tuberous root ofAconitum carmichaeli Debeaux or Aconitumjaponicum Thunberg (Ranunculaceae) prepared bythe following processes.
Process 1: Autoclaving. [Processed Aconite Root 1]Process 2: Heating or autoclaving after rinsing in
the salt, rock salt or calcium chloride water. [Proc-essed Aconite Root 2]
Process 3: Treating with lime after rinsing in the saltwater. [Processed Aconite Root 3]
There are three kinds of Processed Aconite Rootbased on their preparing processes as shown above.
Processed Aconite Root 1, Processed Aconite Root2 and Processed Aconite Root 3 contain the totalalkaloid [as benzoylaconine (C32H45NO10: 603.70)] ofnot less than 0.7z and not more than 1.5z, not lessthan 0.1z and not more than 0.6z, and not less than0.5z and not more than 0.9z, calculated on the driedbases, respectively.
The label indicates the treating process.
DescriptionProcessed Aconite Root 1: Cut pieces irregularly polygonal,less than 10 mm in diameter; externally dark grayish brownto blackish brown; hard in texture; cut surface ‰at, lightbrown to dark brown, usually horny and lustrous.
Odor, weak and characteristic.Under a microscope, transverse and longitudinal sections
reveal pitted, scaraliform, reticulate and spiral vessels;starch grains in parenchymatous cells usually gelatinized butsometimes not gelatinized; starch grains, simple, spherical orellipsoid, 2 – 25 mm in diameter, or 2- to a dozen or so- com-pound, hilum of starch grain distinct.
Processed Aconite Root 2: Nearly obconical root, 15 – 30mm in length, 12 – 16 mm in diameter, slices cut longitudi-nally or transversely, 20 – 60 mm in length, 15 – 40 mm inwidth, and 200 – 700 mm in thickness, or cut pieces irregular-ly polygonal, less than 12 mm in diameter; externally lightbrown to dark brown or yellowish brown; hard in texture,usually without wrinkles; cut surface ‰at, light brown todark brown or yellowish white to light yellowish brown, usu-ally horny, semi-transparent and lustrous.
Odor, weak and characteristic.
17711771Supplement II, JPXIV O‹cial Monographs for Part II
Under a microscope, transverse and longitudinal sectionsreveal metaderm, primary cortex, endodermis, secondarycortex, cambium, and xylem; primary cortex containsoblong to oblong-square sclerenchymatous cells, 30 – 75 mmin short axis, 60 – 150 mm in long axis; endodermis singlelayered, endodermal cells elongated in tangential direction;cambium, star shaped or irregular polygons to orbicular; agroup of vessel in xylem v-shaped; sometimes isolated ringof cambium appears in secondary cortex or in pith; vessels,pitted, scaraliform, reticulate and spiral; starch grains inparenchymatous cells gelatinized.
Processed Aconite Root 3: Cut pieces irregularly polygonal,less than 5 mm in diameter; externally grayish brown; hardin texture; cut surface ‰at, light grayish brown to grayishwhite, not lustrous.
Odor, weak and characteristic.Under a microscope, transverse and longitudinal sections
reveal pitted, scaraliform, reticulate and spiral vessels;starch grains, simple, spherical or ellipsoid, 2 – 25 mm indiameter, or 2- to a dozen or so- compound, hilum of starchgrain distinct.
Identiˆcation To 3 g of pulverized Processed Aconite Rootadd 20 mL of diethyl ether and 2 mL of ammonia TS, shakefor 10 minutes, and centrifuge. Evaporate the ether layer todryness under reduced pressure, dissolve the residue in 1 mLof diethyl ether, and use this solution as the sample solution.Separately, dissolve 1 mg of benzoylmesaconine hydrochlo-ride for thin-layer chromatography in 10 mL of ethanol(99.5), and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography, develop the plate with a mixtureof ethyl acetate, ethanol (99.5) and ammonia water (28)(40:3:2) to a distance of about 10 cm, and air-dry the plate.Spray evenly DragendorŠ's TS for spraying on the plate,air-dry the plate, and spray evenly sodium nitrite TS: one ofthe spot among the several spots from the sample solutionhas the same color tone and Rf value with the yellow-brownspot from the standard solution.
Purity Aconitum diester alkaloids (aconitine, jesaconitine,hypaconitine and mesaconitine)—Weigh accurately about0.5 g of pulverized Processed Aconite Root, put in a glass-stoppered centrifuge tube, suspend in 3.0 mL of water byshaking, and add 1.0 mL of ammonia TS and 20 mL ofdiethyl ether. Stopper tightly the tube, shake for 30 minutes,centrifuge, and separate the ether layer. To the residue add1.0 mL of ammonia TS and 20 mL of diethyl ether, andrepeat the above process two times. Combine all extracts,evaporate to dryness under reduced pressure at not morethan 409C, and dissolve the residue with exactly 10 mL of amixture of phosphate buŠer solution for aconite root andacetonitrile (1:1). Centrifuge this solution, and use thesupernatant liquid as the sample solution. Perform the testwith exactly 20 mL each of the sample solution and the aconi-tum diester alkaloids standard solution as directed under the
Liquid Chromatography according to the followingconditions, and determine the heights of the peaks corre-sponding to aconitine, jesaconitine, hypaconitine andmesaconitine, HTA and HSA, HTJ and HSJ, HTH and HSH, andHTM and HSM, respectively, and calculate the amounts ofthem by the following formulae: the amounts of aconitine,jesaconitine, hypaconitine and mesaconitine per g calculatedon the dried basis are not more than 60 mg, 60 mg, 280 mg and140 mg, respectively, and the total amount of them is notmore than 450 mg.
Amount (mg) of aconitine (C34H47NO11)
＝CSA
W×
HTA
HSA×10
Amount (mg) of jesaconitine (C35H49NO12)
＝CSJ
W×
HTJ
HSJ×10
Amount (mg) of hypaconitine (C33H45NO10)
＝CSH
W×
HTH
HSH×10
Amount (mg) of mesaconitine (C33H45NO11)
＝CSM
W×
HTM
HSM×10
CSA: Concentration (mg/mL) of aconitine for purity in theaconitum diester alkaloids standard solution forpurity
CSJ: Concentration (mg/mL) of jesaconitine for purity inthe aconitum diester alkaloids standard solution forpurity
CSH: Concentration (mg/mL) of hypaconitine for purity inthe aconitum diester alkaloids standard solution forpurity
CSM: Concentration (mg/mL) of mesaconitine for purity inthe aconitum diester alkaloids standard solution forpurity
W: Amount (g) of the sample, calculated on the driedbasis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 231 nm for aconitine, hypaconitine andmesaconitine; 254 nm for jesaconitine).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: A mixture of phosphate buŠer solution foraconite root and tetrahydrofuran (183:17).
Flow rate: Adjust the ‰ow rate so that the retention timeof mesaconitine is about 31 minutes.System suitability—
System performance: When the procedure is run with20 mL of the aconitum diester alkaloids standard solutionfor purity under the above operating conditions, using 254
17721772 Supplement II, JPXIVO‹cial Monographs for Part II
nm, mesaconitine, hypaconitine, aconitine and jesaconitineare eluted in this order, and each resolution between theirpeaks is not less than 1.5, respectively.
System repeatability: To 1 mL of aconitum diesteralkaloids standard solution for purity add a mixture of phos-phate buŠer solution for aconite root and acetonitrile (1:1)to make 10 mL. When the test is repeated 6 times with 20 mLof this solution under the above operating conditions, using231 nm, the relative standard deviation of the peak height ofmesaconitine is not more than 1.5z.
Loss on drying Not more than 15.0z (6 hours).
Total ashProcessed Aconite Root 1: Not more than 4.0z.Processed Aconite Root 2: Not more than 12.0z.Processed Aconite Root 3: Not more than 19.0z.
Acid-insoluble ash Not more than 0.9z.
Assay Weigh accurately about 2 g of pulverized ProcessedAconite Root, put in a glass-stoppered centrifuge tube, andadd 1.6 mL of ammonia TS and 20 mL of diethyl ether.Stopper tightly the tube, shake for 30 minutes, centrifuge,and separate the ether layer. To the residue add 0.8 mL ofammonia TS and 20 mL of diethyl ether, and proceed asabove. Repeat this process more three times. Combine allextracts, evaporate to dryness under reduced pressure,dissolve the residue in 5 mL of ethanol (99.5), add 30 mL offreshly boiled and cooled water, and titrate with 0.01 mol/Lhydrochloric acid VS until the color of the solution changesfrom green to gray-blue through blue-green (indicator: 3drops of methyl red-methylene blue TS). Perform a blankdetermination and make any necessary correction.
Each mL of 0.01 mol/L hydrochloric acid VS＝6.037 mg of total alkaloid [as benzoylaconine
Powdered Processed Aconite Root is the powder ofProcessed Aconite Root prepared by Process 1 orProcess 2, or the powder of the tuberous root ofAconitum carmichaeli Debeaux or Aconitum japoni-cum Thunberg (Ranunculaceae) prepared by Process1.
Occasionally, it contains Corn Starch or Lactose.Process 1: Autoclaving. [Powdered Processed
Aconite Root 1]Process 2: Heating or autoclaving after rinsing in
the salt, rock salt or calcium chloride water. [Pow-dered Processed Aconite Root 2]
There are two kinds of Powdered Processed Aconite
Root based on their preparing processes as shownabove.
Powdered Processed Aconite Root 1 and PowderedProcessed Aconite Root 2 contain the total alkaloid[as benzoylaconine (C32H45NO10: 603.70)] of not lessthan 0.4z and not more than 1.2z, and not less than0.1z and not more than 0.3z, calculated on the driedbases, respectively.
The label indicates the treating process.
DescriptionPowdered Processed Aconite Root 1: Powdered ProcessedAconite Root 1 occurs as a light grayish brown powder. Ithas a characteristic odor.
Under a microscope, Powered Processed Aconite Root 1reveals gelatinized starch masses or starch grains and paren-chymatous cells containing them, fragments of reddishbrown metaderm, fragments of pitted, scaraliform, reticu-late and spiral vessels; also square to oblong-square scleren-chymatous cells, 30 – 150 mm in diameter, 100 – 250 mm inlength, cell wall of sclerenchymatous cells, 6 – 12 mm inthickness; starch grains of ``Processed Aconite Root'', sim-ple, spherical or ellipsoid, 2 – 25 mm in diameter, or 2- to adozen or so- compound, hilum of starch grain distinct.
Powdered Processed Aconite Root 2: Powdered ProcessedAconite Root 2 occurs as a light yellowish white powder. Ithas a characteristic odor.
Under a microscope, Powered Processed Aconite Root 2reveals gelatinized starch masses and parenchymatous cellscontaining them, fragments of reddish brown metaderm,fragments of pitted, scaraliform, reticulate and spiralvessels; also square to oblong-square sclerenchymatous cells,30 – 150 mm in diameter, 100 – 250 mm in length, cell wall ofsclerenchymatous cells, 6 – 12 mm in thickness.
Identiˆcation To 3 g of Powdered Processed Aconite Rootadd 2 mL of ammonia TS and 20 mL of diethyl ether, shakefor 10 minutes, and centrifuge. Evaporate the ether layer todryness under reduced pressure, dissolve the residue in 1 mLof diethyl ether, and use this solution as the sample solution.Separately, dissolve 1 mg of benzoylmesaconine hydrochlo-ride for thin-layer chromatography in 10 mL of ethanol(99.5), and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography, develop the plate with a mixtureof ethyl acetate, ethanol (99.5) and ammonia water (28)(40:3:2) to a distance of about 10 cm, and air-dry the plate.Spray evenly DragendorŠ's TS for spraying on the plate,air-dry the plate, and spray evenly sodium nitrite TS: one ofthe spot among the several spots from the sample solutionhas the same color tone and Rf value with the yellow-brownspot from the standard solution.
Purity Aconitum diester alkaloids (aconitine, jesaconitine,hypaconitine and mesaconitine)—Weigh accurately about0.5 g of Powdered Processed Aconite Root, put in a glass-stoppered centrifuge tube, suspend in 3.0 mL of water by
17731773Supplement II, JPXIV O‹cial Monographs for Part II
shaking, and add 1.0 mL of ammonia TS and 20 mL ofdiethyl ether. Stopper tightly the tube, shake for 30 minutes,centrifuge, and separate the ether layer. To the residue add1.0 mL of ammonia TS and 20 mL of diethyl ether, andrepeat the above process two times. Combine all extracts,evaporate to dryness under reduced pressure at not morethan 409C, and dissolve the residue with exactly 10 mL of amixture of phosphate buŠer solution for aconite root andacetonitrile (1:1). Centrifuge this solution, and use thesupernatant liquid as the sample solution. Perform the testwith exactly 20 mL each of the sample solution and the aconi-tum diester alkaloids standard solution as directed under theLiquid Chromatography according to the followingconditions, and determine the heights of the peaks corre-sponding to aconitine, jesaconitine, hypaconitine andmesaconitine, HTA and HSA, HTJ and HSJ, HTH and HSH, andHTM and HSM, respectively, and calculate the amounts ofthem by the following formulae: the amounts of aconitine,jesaconitine, hypaconitine and mesaconitine per g calculatedon the dried basis are not more than 55 mg, 40 mg, 55 mg and120 mg, respectively, and the total amount of them is notmore than 230 mg.
Amount (mg) of aconitine (C34H47NO11)
＝CSA
W×
HTA
HSA×10
Amount (mg) of jesaconitine (C35H49NO12)
＝CSJ
W×
HTJ
HSJ×10
Amount (mg) of hypaconitine (C33H45NO10)
＝CSH
W×
HTH
HSH×10
Amount (mg) of mesaconitine (C33H45NO11)
＝CSM
W×
HTM
HSM×10
CSA: Concentration (mg/mL) of aconitine for purity in theaconitum diester alkaloids standard solution forpurity
CSJ: Concentration (mg/mL) of jesaconitine for purity inthe aconitum diester alkaloids standard solution forpurity
CSH: Concentration (mg/mL) of hypaconitine for purity inthe aconitum diester alkaloids standard solution forpurity
CSM: Concentration (mg/mL) of mesaconitine for purity inthe aconitum diester alkaloids standard solution forpurity
W: Amount (g) of the sample, calculated on the driedbasis
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 231 nm for aconitine, hypaconitine andmesaconitine; 254 nm for jesaconitine).
Column: A stainless steel column 4.6 mm in insidediameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in
particle diameter).Column temperature: A constant temperature of about
409C.Mobile phase: A mixture of phosphate buŠer solution for
aconite root and tetrahydrofuran (183:17).Flow rate: Adjust the ‰ow rate so that the retention time
of mesaconitine is about 31 minutes.System suitability—
System performance: When the procedure is run with 20mL of the aconitum diester alkaloids standard solution forpurity under the above operating conditions, using 254 nm,mesaconitine, hypaconitine, aconitine and jesaconitine areeluted in this order, and each resolution between their peaksis not less than 1.5, respectively.
System repeatability: To 1 mL of aconitum diesteralkaloids standard solution for purity add a mixture ofphosphate buŠer solution for aconite root and acetonitrile(1:1) to make 10 mL. When the test is repeated 6 times with20 mL of this solution under the above operating conditions,using 231 nm, the relative standard deviation of the peakheight of mesaconitine is not more than 1.5z.
Loss on drying Not more than 11.0z (6 hours).
Total ashPowdered Processed Aconite Root 1: Not more than
4.0z.Powdered Processed Aconite Root 2: Not more than
7.0z.
Acid-insoluble ash Not more than 0.7z.
Assay Weigh accurately about 2 g of Powdered ProcessedAconite Root, put in a glass-stoppered centrifuge tube, andadd 1.6 mL of ammonia TS and 20 mL of diethyl ether.Stopper tightly the tube, shake for 30 minutes, centrifuge,and separate the ether layer. To the residue add 0.8 mL ofammonia TS and 20 mL of diethyl ether, and proceed asabove. Repeat this process more three times. Combine allextracts, evaporate to dryness under reduced pressure,dissolve the residue in 5 mL of ethanol (99.5), add 30 mL offreshly boiled and cooled water, and titrate with 0.01 mol/Lhydrochloric acid VS until the color of the solution changesfrom green to gray-blue through blue-green (indicator: 3drops of methyl red-methylene blue TS). Perform a blankdetermination and make any necessary correction.
Each mL of 0.01 mol/L hydrochloric acid VS＝6.037 mg of total alkaloid [as benzoylaconine
(C32H45NO10: 603.70)]
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Add the following:
Processed Ginger
Zingiberis Processum Rhizoma
カンキョウ
Processed Ginger is the rhizome of Zingibero‹cinale Roscoe (Zingiberaceae), after being passedthrough hot water or being steamed.
Description Irregularly compressed and often branchedmassive rhizome; branched parts slightly curved ovoid oroblong-ovoid, 2 – 4 cm in length, and 1 – 2 cm in diameter;external surface grayish yellow to grayish yellow-brown,with wrinkles and ring node; fractured surface brown todark brown, transparent and horny; under a magnifyingglass, a transverse section reveals cortex and stele distinctlydivided; vascular bundles scattered throughout the surface.
Odor, characteristic; taste, extremely pungent.Under a microscope, a transverse section reveals cork
layer, cortex and stele in this order from the outside; cortexand stele, divided by a single-layered endodermis, composedof parenchyma, vascular bundles scattered and surroundedby ˆber bundles; oil cells contain yellow oil-like substances,scattered in parenchyma; parenchymatous cells containsolitary crystals of calcium oxalate, and gelatinized starch.
Identiˆcation To 2 g of pulverized Processed Ginger add 5mL of diethyl ether, shake for 10 minutes, ˆlter, and use theˆltrate as the sample solution (1). To the residue add 5 mL ofmethanol, proceed in the same manner as above, and use soobtained solution as the sample solution (2). Separately,dissolve 1 mg of [6]-shogaol for thin-layer chromatographyin 2 mL of methanol, and use this solution as the standardsolution (1). Separately, dissolve 1 mg of Sucrose in 2 mL ofmethanol, and use this solution as the standard solution (2).Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution (1) and the standard solution (1) on a plate of silicagel for thin-layer chromatography. Develop the plate with amixture of ethyl acetate and hexane (1:1) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate, heat at 1059Cfor 5 minutes, and allow to cool: one of the spot among theseveral spots from the sample solution (1) has the same colortone and Rf value with the green spot from the standardsolution (1). Spot 10 mL each of the sample solution (2) andthe standard solution (2) on a plate of silica gel for thin-layerchromatography, develop the plate with a mixture of 1-butanol, water and acetic acid (100) (8:5:3) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly 1,3-naphthalenediol TS on the plate, and heat at 1059C for 5minutes: one of the spot among the several spots from thesample solution (2) has the same color tone and Rf valuewith the red-purple spot from the standard solution (2).
Loss on drying Not more than 15.0z (6 hours).
Total ash Not more than 6.5z.
Acid-insoluble ash Not more than 1.5z.
Extract content Not less than 8.0z (dilute ethanol-solubleextract).
Rhubarbダイオウ
Change the origin/limits of content to read:
Rhubarb is usually the rhizome of Rheum palmatumLinn áe, Rheum tanguticum Maximowicz, Rheumo‹cinale Baillon, Rheum coreanum Nakai or theirinterspeciˆc hybrids (Polygonaceae).
It contains not less than 0.25z of sennosides A(C42H38O20: 862.74), calculated on the basis of driedmaterial.
Powdered Rhubarbダイオウ末
Change the origin/limits of content to read:
Powdered Rhubarb is the powder of Rhubarb.It contains not less than 0.25z of sennoside A
(C42H38O20: 862.74), calculated on the basis of driedmaterials.
Add the following:
Sappan Wood
Sappan Lignum
ソボク
Sappan Wood is the duramen of Caesalpinia sappanLinn áe (Leguminosae).
Description Chips, slices or short pieces of wood; yellow-ish red to grayish yellow-brown, sometimes with light brownto grayish white splint woods; hard in texture; a transversesection shows a pattern like annual ring.
Almost odorless; almost tasteless.Under a microscope, a transverse section reveals ray
composed of 1 – 2 rows of slender and long cells; the areabetween rays ˆlled with ˆber cells, and large and oblongvessels scattered there; solitary crystals of calcium oxalate inparenchymatous cells of the innermost of xylem.
Identiˆcation To 0.5 g of pulverized Sappan Wood add 10mL of dilute ethanol, shake, and ˆlter. To 5 mL of theˆltrate add 2 to 3 drops of sodium hydroxide TS: a dark red
17751775Supplement II, JPXIV O‹cial Monographs for Part II
color develops.
Purity Put a small piece of Sappan Wood in calciumhydroxide TS: no purple-blue color develops.
Loss on drying Not more than 11.5z (6 hours).
Total ash Not more than 2.0z.
Extract content Not less than 7.0z (dilute ethanol-solubleextract).
Powdered Scutellaria Rootオウゴン末
Change the origin/limits of content to read:
Powdered Scutellaria Root is the powder of Scutel-laria Root.
It contains not less than 10.0z of baicalin(C21H18O11: 446.36), calculated on the basis of driedmaterial.
Add the following:
Termeric
Curcumae Rhizoma
ウコン
Termeric is the rhizome or being removed the corklayer from it of Curcuma longa Linn áe (Zingiberaceae),usually after being passed through hot water.
Description Termeric is a main rhizome or a lateral rhi-zome; main rhizome, nearly ovoid, about 3 cm in diameter,about 4 cm in length; lateral rhizome, cylindrical, withround tips, curved, about 1 cm in diameter, 2 - 6 cm inlength; both main and lateral rhizomes with cyclic nodes;rhizome with cork layer, yellowish brown, lustrous; rhizomewithout cork layer, dark yellowish red, with yellowish redpowders on surface; hard in texture, not easily broken;transversely cut surface yellowish brown to reddish brown,lustrous like wax.
Odor, characteristic; taste, slightly bitter and stimulant, itcolors a saliva yellow on chewing.
Under a microscope, a transverse section reveals theoutermost layer to be composed of a cork layer 4 – 10 cellsthick; sometimes a cork layer partly remains; cortex andstele, divided by a single-layered endodermis, composed ofparenchyma, vascular bundles scattered; oil cells scattered inparenchyma; parenchymatous cells contain yellow sub-stances, sandy and solitary crystals of calcium oxalate, andgelatinized starch.
Identiˆcation To 0.5 g of pulverized Termeric add 20 mLof methanol, shake for 15 minutes, ˆlter and use the ˆltrate
as the sample solution. Perform the test with this solution asdirected under the Thin-layer Chromatography. Spot 5 mLof the sample solution on a plate of silica gel for thin-layerchromatography. Develop the plate with a mixture of ethylacetate, hexane and acetic acid (100) (70:30:1) to a distanceof about 10 cm, and air-dry the plate: a yellow spot appearsat around Rf 0.4.
Loss on drying Not more than 17.0z (6 hours).
Total ash Not more than 7.5z.
Acid-insoluble ash Not more than 1.0z.
Extract content Not less than 9.0z (dilute ethanol-solubleextract).
Add the following:
Tribulus Fruit
Tribuli Fructus
シツリシ
Tribulus Fruit is the fruit of Tribulus terrestrisLinn áe (Zygophyllaceae).
Description Pentagonal star shaped fruit, composed of ˆvemericarps, 7 – 12 mm in diameter, often each mericarpseparated; externally grayish green to grayish brown; a pairof longer and shorter spines on surface of each mericarp, thelonger spine 3 – 7 mm in length, the shorter one 2 – 5 mm inlength, numerous small processes on midrib; pericarp hardin texture, cut surface light yellow; each mericarp contains 1– 3 seeds.
Almost odorless; taste, mild at ˆrst, followed by bitter-ness.
Under a microscope, a transverse section reveals epicarpcomposed of a single-layered epidermis; mesocarp com-posed of parenchyma and sclerenchyma layer; endocarpcomposed of several-layered ˆber cells; a single-layer of cellbetween mesocarp and endocarp contain solitary crystals ofcalcium oxalate; cotyledons of seed contain oil drops andaleurone grains, and occasionally starch grains.
Identiˆcation To 2 g of pulverized Tribulus Fruit add 5 mLof methanol, shake for 10 minutes, ˆlter, and use the ˆltrateas the sample solution. Perform the test with this solution asdirected under the Thin-layer Chromatography. Spot 10 mLof the sample solution on a plate of silica gel for thin-layerchromatography, develop the plate with a mixture of ethylacetate and water (40:1) to a distance of about 10 cm, andair-dry the plate. Spray evenly dilute sulfuric acid on theplate, heat at 1059C for 5 minutes, and examine underultraviolet light (main wavelength: 365 nm): a blue-white‰uorescent spot appears at around Rf 0.4.
Purity (1) Peduncle—Not more than 4.0z.(2) Foreign matters—Not more than 1.0z of foreign
17761776 Supplement II, JPXIVO‹cial Monographs for Part II
matters other than peduncle.
Loss on drying Not more than 11.0z (6 hours).
Total ash Not more than 13.0z.
Acid-insoluble ash Not more than 1.5z.
Extract content Not less than 8.5z (dilute ethanol-solubleextract).
Uncaria Thornチョウトウコウ
Change the origin/limits of the content to read:
Uncaria Thorn is, usually the prickle, of Uncariarhynchophylla Miquel, Uncaria sinensis Haviland orUncaria macrophylla Wallich (Rubiaceae).
Uncaria Thorn contains not less than 0.03z of totalalkaloids (rhynchophylline and hirstine), calculated onthe dried basis.
Change the Component determination to read:
Component determination Weigh accurately about 0.2 gof medium powdered Uncaria Thorn, transfer into a glass-stoppered centrifuge tube, add 30 mL of a mixture ofmethanol and dilute acetic acid (7:3), shake for 30 minutes,centrifuge, and separate the supernatant liquid. To theresidue add two 10-mL portions of a mixture of methanoland dilute acetic acid (7:3), proceed in the same manner, andcombine all of the supernatant liquid. To the combinedliquid add a mixture of methanol and dilute acetic acid (7:3)to make exactly 50 mL, and use this as the sample solution.Separately, weigh accurately about 5 mg of rhynchophyllinefor component determination, previously dried in a desicca-tor (silica gel) for 24 hours, and dissolve in a mixture ofmethanol and dilute acetic acid (7:3) to make exactly 100mL. Pipet 1 mL of this solution, add a mixture of methanoland dilute acetic acid (7:3) to make exactly 10 mL, and usethis solution as the standard solution (1). Separately, dis-solve 1 mg of hirsutine in 100 mL of a mixture of methanoland dilute acetic acid (7:3), and use this solution as the stan-dard solution (2). Perform the test with exactly 20 mL eachof the sample solution and the standard solution (1) and (2)as directed under the Liquid Chromatography according tothe following conditions, and determine the peak areas, ATa
and ATb, of rhynchophylline and hirsutine obtained from thesample solution, and the peak area, AS, of rhynchophyllinefrom the standard solution (1).
Amount (mg) of total alkaloids (rhynchophylline andhirstine)
＝WS×ATa＋1.405ATb
AS×
120
WS: Amount (mg) of rhynchophylline for componentdetermination
Operating conditions—Detector: An ultraviolet absorption photometer
(wavelength: 245 nm).Column: A stainless steel column 4.6 mm in inside
diameter and 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm inparticle diameter).
Column temperature: A constant temperature of about409C.
Mobile phase: Dissolve 3.85 g of ammonium acetate in200 mL of water, add 10 mL of acetic acid (100) and waterto make 1000 mL, and add 350 mL of acetonitrile.
Flow rate: Adjust the ‰ow rate so that the retention timeof rhynchophylline is about 17 minutes.System suitability—
System performance: Dissolve 5 mg of rhynchophyllinefor component determination in 100 mL of a mixture ofmethanol and dilute acetic acid (7:3). To 5 mL of this solu-tion add 1 mL of ammonia solution (28), and re‰ux for 10minutes or warm at about 509C for 2 hours. After cooling,to 1 mL of the solution so obtained add a mixture ofmethanol and dilute acetic acid (7:3) to make 5 mL. Whenthe procedure is run with 20 mL of this solution under theabove operating conditions, the peak of isorhynchophyllineis appears in addition to the peak of rhynchophylline, andthe resolution between these peaks is not less than 1.5.
System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution (1) under the aboveoperating conditions, the relative standard deviation of thepeak areas of rhynchophylline is not more than 1.5z.
Change to read:
Wheat Starch
Amylum Tritici
コムギデンプン
Wheat Starch consists of the starch granulesobtained from the seeds of Triticum aestivum Linn áe(Gramineae).
Description Wheat Starch occurs as white masses orpowder.
It is practically insoluble in water and in ethanol (99.5).
Identiˆcation (1) Under a microscope, Wheat Starch,preserved in a mixture of water and glycerin (1:1), appears aslarge and small sized simple grains, or quite rarely mediansized simple grains; usually, large sized grains usually 10 – 60mm in diameter, from upper view, disc like or quite rarelyreniform, centric hilum and striation indistinct or hardlydistinct, often cleft on marginal portion visible; from lateralview, narrowly ellipsoid or fusiform, hilum recognized as acleft along with long axis; small sized grains 2 – 10 mm indiameter, spherical or polygonal; a black cross, its intersec-tion point on hilum, is observed when grains are put between
17771777Supplement II, JPXIV O‹cial Monographs for Part II
two polarizing prisms ˆxed at right angle to each other.(2) To 1 g of Wheat Starch add 50 mL of water, boil for
1 minute, and allow to cool: a subtle white-turbid, pastyliquid is formed.
(3) To 1 mL of the pasty liquid obtained in (2) add 0.05mL of iodine TS: a dark blue-purple color is formed, and thecolor disappears by heating.
pH Put 5.0 g of Wheat Starch in a non-metal vessel, add25.0 mL of freshly boiled and cooled water, mix gently for 1minute, and allow to stand for 15 minutes: the pH of thesolution is between 4.5 and 7.0.
Purity (1) Iron—To 1.5 g of Wheat Starch add 15 mL of2 mol/L hydrochloric acid TS, mix, ˆlter, and use the ˆltrateas the test solution. To 2.0 mL of Standard Iron Solutionadd water to make 20 mL, and use as the control solution.Put 10 mL each of the test solution and the control solutionin test tubes, add 2 mL of a solution of citric acid (2 in 10)and 0.1 mL of mercapto acetic acid, and mix. Alkalize withammonia solution (28) to litmus paper, add water to make20 mL, and mix. Transfer 10 mL each of these solutions intotest tubes, allow to stand for 5 minutes, and compare thecolor of these solutions against a white background: thecolor of the test solution is not darker than that of thecontrol solution (not more than 10 ppm).
(2) Oxidizing substances—To 4.0 g of Wheat Starch add50.0 mL of water, shake for 5 minutes, and centrifuge. To30.0 mL of the supernatant liquid add 1 mL of acetic acid(100) and 0.5 to 1.0 g of potassium iodide, shake, and allowto stand for 25 to 30 minutes at a dark place. Add 1 mL ofstarch TS, and titrate with 0.002 mol/L sodium thiosulfateVS until the color of the solution disappears. Perform ablank determination and make any necessary correction: thevolume of 0.002 mol/L sodium thiosulfate VS consumed isnot more than 1.4 mL (not more than 20 ppm, calculated ashydrogen peroxide).
(3) Sulfur dioxide—(i) Apparatus Use as shown in the following ˆgure.
(ii) Procedure Introduce 150 mL of water into the boil-ing ‰ask, close the tap of the funnel, and pass carbondioxide through the whole system at a rate of 100±5 mL perminute. Pass cooling water through the condenser, and place10 mL of hydrogen peroxide-sodium hydroxide TS in thetest-tube. After 15 minutes, remove the funnel without inter-rupting the stream of carbon dioxide, and introduce throughthe opening into the ‰ask about 25 g of Wheat Starch,accurately weighed, with the aid of 100 mL of water. Applytap grease to the outside of the connection part of thefunnel, and load the funnel. Close the tap of the funnel,pour 80 mL of 2 mol/L hydrochloric TS acid into thefunnel, open the tap to introduce the hydrochloric acid intothe ‰ask, and close the tap while several mL of thehydrochloric acid remains, in order to avoid losing sulfurdioxide. Place the ‰ask in a water bath, and heat the mixturefor 1 hour. Transfer the contents of the test-tube with the aidof a little water to a wide-necked conical ‰ask. Heat in awater bath for 15 minutes, and cool. Add 0.1 mL ofbromophenol blue TS, and titrate with 0.1 mol/L sodiumhydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determi-nation and make any necessary correction. Calculate theamount of sulfur dioxide by applying the following formula:it is not more than 50 ppm.
Amount (ppm) of sulfur dioxide＝ VW×1000×3.203
W: Amount (g) of the sampleV: Amount (mL) of 0.1 mol/L sodium hydroxide VS
consumed
Loss on drying Not more than 15.0z (1 g, 1309C, 90minutes).
Residue on ignition Not more than 0.6z (1 g).
Containers and storage Containers—Well-closed contain-ers.
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Delete the following Ultraviolet-visible Reference Spectra:
Part I
Furosemide 1
Furosemide 2
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Page citations refer to the pages of the Supplement I, and to pages ofthe JP XIV main volume, including those where the text being revised inthe Supplement originally appeared. This Supplement II commenceswith page 1669 and succeeding Supplements will continue to be paged insequence.
1„1358 Main Volume of JP XIV1359„1668 Supplement I1669„1866 Supplement II
