ANTIBIOTIC DRUGS Subpart Aâ€”Bulk Drugs - GPO

865

Food and Drug Administration, HHS § 449.4

this chapter, preparing the sample forassay as follows: Place an accuratelyweighed representative portion of thesample into a high-speed glass blenderjar containing 1.0 milliliter poly-sorbate 80 and sufficient 10 percent po-tassium phosphate buffer, pH 6.0 (solu-tion 6), to obtain a stock solution ofconvenient concentration. Blend for 3to 5 minutes. Further dilute an aliquotof the stock solution with solution 6 tothe reference concentration of 10 unitsof polymyxin B per milliliter (esti-mated).

(2) Sterility. Proceed as directed in§ 436.20(e)(1) of this chapter, except dis-solve the ointment as follows:Aseptically transfer a portion of 0.25gram from each of 10 immediate con-tainers of the drug to 400 milliliters ofdiluting fluid D in an Erlenmeyerflask. Repeat the procedure on another10 immediate containers. Swirl theflasks to dissolve the ointment.

(3) pH. Proceed as directed in § 436.202of this chapter, using the undilutedsample.

[39 FR 19115, May 30, 1974, as amended at 46FR 16684, Mar. 13, 1981; 50 FR 19920, May 13,1985]

PART 449—ANTIFUNGALANTIBIOTIC DRUGS

Subpart A—Bulk Drugs

Sec.449.1—449.3 [Reserved]449.4 Amphotericin B.449.4a Amphotericin B for use in parenteral

products.449.10 Candicidin.449.20 Griseofulvin.449.40 Natamycin.449.50 Nystatin.

Subpart B—Oral Dosage Forms

449.104 Amphotericin B oral suspension.449.120 Griseofulvin oral dosage forms.449.120a Griseofulvin tablets.449.120b Griseofulvin capsules.449.120c Griseofulvin oral suspension.449.120d Griseofulvin (ultramicrosize) tab-

lets.449.150 Nystatin oral dosage forms.449.150a Nystatin tablets.449.150b Nystatin oral suspension.449.150c Nystatin for oral suspension.449.150d Nystatin pastilles.

Subpart C—Injectable Dosage Forms

449.204 Amphotericin B for injection.

Subpart D—Ophthalmic Dosage Forms

449.340 Natamycin ophthalmic suspension.

Subpart E [Reserved]

Subpart F—Dermatologic Dosage Forms

449.504 Amphotericin B dermatologic dosageforms.

449.504a Amphotericin B ointment.449.504b Amphotericin B cream.449.504c Amphotericin B lotion.449.550 Nystatin dermatologic dosage forms.449.550a Nystatin ointment.449.550b Nystatin-iodochlorhydroxyquin

ointment.449.550c Nystatin-neomycin sulfate-grami-

cidin-triamcinolone acetonide ointment;nystatin-neomycin sulfate-gramicidin-fludrocortisone acetate ointment.

449.550d Nystatin cream.449.550e Nystatin-neomycin sulfate-grami-

cidin-triamcinolone acetonide cream.449.550f Nystatin topical powder.449.550g Nystatin-neomycin sulfate-grami-

cidin topical powder.449.550h Nystatin lotion.

Subpart G—Vaginal Dosage Forms

449.610 Candicidin vaginal dosage forms.449.610a Candicidin vaginal ointment.449.610b Candicidin vaginal tablets.449.610c Candicidin vaginal capsules.449.650 Nystatin vaginal dosage forms.449.650a Nystatin vaginal tablets.449.650b Nystatin vaginal suppositories.

AUTHORITY: 21 U.S.C. 357.

SOURCE: 39 FR 19134, May 30, 1974, unlessotherwise noted.


§§ 449.1—449.3 [Reserved]

§ 449.4 Amphotericin B.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Amphotericin B is a yellowto golden-orange powder. It is insolublein water at pH. 6.0 to 7.0, anhydrous al-cohols, esters, ethers, benzene, and tol-uene. It is soluble indimethylformamide anddimethylsulfoxide. It is so purified anddried that:

(i) Its potency is not less than 750micrograms of amphotericin B per mil-ligram on an anhydrous basis.

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21 CFR Ch. I (4–1–98 Edition)§ 449.4

(ii) It contains not more than 15 per-cent of amphotericin A.

(iii) [Reserved](iv) Its loss on drying is not more

than 5.0 percent.(v) It contains not more than 3.0 per-

cent residue on ignition.(vi) It passes the identity test.(2) Labeling. In addition to the label-

ing prescribed by § 432.5(b) of this chap-ter, each package shall bear on itslabel the statements ‘‘Store below 10°C.’’ and ‘‘Protect from light and mois-ture’’.

(3) Requests for certification; samples.In addition to the requirements of§ 431.1 of this chapter, each such re-quest shall contain:

(i) Results of test and assays on thebatch for potency, amphotericin A con-tent, loss on drying, residue oniginition, and identity.

(ii) Samples required on the batch: 10packages, each containing not lessthan 500 milligrams.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Dissolve an accu-rately weighed sample in sufficientdimethylsulfoxide to give a stock solu-tion of convenient concentration. Fur-ther dilute an aliquot withdimethylsulfoxide to a concentrationof 20 micrograms of amphotericin B permilliliter (estimated). Remove an ali-quot; dilute with 0.2M potassium phos-phate buffer, pH 10.5 (solution 10), tothe reference concentration of 1.0microgram of amphotericin B per mil-liliter (estimated).

(2) Amphotericin A content—(i)Amphotericin A. Dry approximately 20milligrams of the nystatin workingstandard as described in § 436.200(a) ofthis chapter. Accurately weigh thedried working standard and quan-titatively transfer into a 200-millilitervolumetric flask. Add exactly 40.0 mil-

liliters of dimethylsulfoxide and dis-solve. Make to mark with methyl alco-hol and mix thoroughly. Pipette 4.0milliliters of this solution into a 50-milliliter volumetric flask. Add methylalcohol to mark and mix thoroughly.

(ii) Amphotericin B. Dry approxi-mately 50 milligrams of theamphotericin B working standard asdescribed in § 436.200(a) of this chapter.Accurately weigh the dried workingstandard and quantitatively transferinto a 50-milliliter volumetric flask.Add 10 milliliters of dimethylsulfoxideand dissolve. Make to mark with meth-yl alcohol and mix thoroughly. Pipette4.0 milliliters of this solution into a 50-milliliter volumetric flask. Add methylalcohol to mark and mix thoroughly.The standard solution should be usedfor 1 day only.

(iii) Sample. Accurately weigh about50 milligrams of the sample to be test-ed and quantitatively transfer into a50-milliliter volumetric flask. Add 10milliliters of dimethylsulfoxide anddissolve. Make to mark with methyl al-cohol and mix thoroughly. Pipette 4.0milliliters of this solution into a 50-milliliter volumetric flask. Add methylalcohol to mark and mix thoroughly.

(iv) Blank. Pipette 10 milliliters ofdimethylsulfoxide into a 50-millilitervolumetric flask. Make to mark withmethyl alcohol and mix. Pipette 4.0milliliters of this solution into a 50-milliliter volumetric flask. Make tomark with methyl alcohol and mixthoroughly.

(v) Procedure. Use a suitable ultra-violet spectrophotometer and 1-centi-meter silica cells. Adjust the instru-ment to zero with the blank solution.Measure the absorbances of the solu-tions of nystatin standard,amphotericin B standard, and the sam-ple at 304 nanometers and at 282nanometers. Calculate the absorptivityof each standard at both wavelengths:

Percent amphotericin A =[(B S b S

W B a) b A

2 1

s

× − × ×

× × − ×

) ( )]

[( ( )]

625

where: A=Absorptivity of nystatin standard at 282nanometers;


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B=Absorptivity of amphotericin B stand-ard at 282 nanometers;

a=Absorptivity of nystatin standard at 304nanometers;

b=Absorptivity of amphotericin B standardat 304 nanometers;

S1=Absorbance of sample at 282nanometers;

S2=Absorbance of sample at 304nanometers;

Ws=Weight of sample in grams (on an an-hydrous basis).

(3) [Reserved](4) Loss on drying. Proceed as directed

in § 436.200(b) of this chapter.(5) Residue on ignition. Proceed as di-

rected in § 436.207(a) of this chapter.(6) Identity. Using the solutions pre-

pared as described in paragraphs (b)(2)(ii), (iii), and (iv) of this section, recordthe absorption spectrum from 320 to 240nanometers. Then dilute these solu-tions (1∂9) with methyl alcohol andrecord the absorption spectrum from400 to 320 nanometers. The sample ex-hibits absorption peaks at identicalwavelengths with that of theamphotericin B standard. Depending onthe amphotericin A content of the sam-ple, a peak may occur at 304nanometers.

[39 FR 19115, May 30, 1974, as amended at 46FR 16684, Mar. 13, 1981; 49 FR 2242, Jan. 19,1984; 50 FR 19920, May 13, 1985]

§ 449.4a Amphotericin B for use in par-enteral products.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Amphotericin B is a yellowto golden-orange powder. It is insolublein water at pH 6.0 to 7.0, anhydrous al-cohols, esters, ethers, benzene, and tol-uene. It is soluble indimethylformamide anddimethylsulfoxide. It is so purified anddried that:

(i) Its potency is not less than 750micrograms of amphotericin B per mil-ligram on an anhydrous basis.

(ii) It contains not more than 5 per-cent of amphotericin A.

(iii) [Reserved](iv) Its loss on drying is not more

than 5.0 percent.(v) It contains not more than 0.5 per-

cent residue on ignition.(vi) It passes the identity test.(2) Labeling. In addition to the label-

ing prescribed by § 432.5(b) of this chap-

ter, each package shall bear on itslabel the statements ‘‘Store below 10°C.’’ and ‘‘Protect from light and mois-ture’’.


(i) Results of tests and assays on thebatch for potency, amphotericin A con-tent, loss on drying, residue on igni-tion, and identity.

(ii) Samples required on the batch: 10packages, each containing not lessthan 500 milligrams.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Dissolve an accu-rately weighed sample in sufficientdimethylsulfoxide to give a stock solu-tion of convenient concentration. Fur-ther dilute with dimethylsulfoxide togive a concentration of 20 microgramsof amphotericin B per milliliter (esti-mated). Dilute an aliquot with 0.2M po-tassium phosphate buffer, pH 10.5 (solu-tion 10), to the reference concentrationof 1.0 microgram of amphotericin B permilliliter (estimated).

(2) Amphotericin A content. Proceed asdirected in § 449.4(b)(2).


in § 436.200(b) of this chapter.(5) Residue of ignition. Proceed as di-

rected in § 436.207(a) of this chapter.(6) Identity. Proceed as directed in

§ 449.4(b)(7).

[39 FR 19134, May 30, 1974, as amended at 49FR 2243, Jan. 19, 1984; 50 FR 19920, May 13,1985]

§ 449.10 Candicidin.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Candicidin is a brown toyellow powder. It is sparingly solublein water; very slightly soluble in ethylalcohol, butyl alcohol, and acetone. Itis so purified and dried that:

(i) Its potency is not less than 1,000micrograms of candicidin per milli-gram on an anhydrous basis.

(ii) Its loss on drying is not morethan 4 percent.

(iii) Its pH is not less than 8.0 normore than 10.0 in a 1 percent aqueoussuspension.


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21 CFR Ch. I (4–1–98 Edition)§ 449.20

(iv) Its ultraviolet absorption spec-trum is characteristic of a conjugatedheptaene and is qualitatively the sameas that of the candicidin workingstandard.

(2) Labeling. It shall be labeled in ac-cordance with the requirements of§ 432.5(b) of this chapter.


(i) Results of tests and assays on thebatch for potency, loss on drying, pH,and identity.

(ii) Samples required: 10 packages,each containing approximately 300 mil-ligrams.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.106 ofthis chapter, preparing the sample forassay as follows: Dissolve a portion ofthe sample in sufficientdimethylsulfoxide to yield an esti-mated concentration of 1,000micrograms of candicidin activity permilliliter. Further dilute an aliquotwith sterile distilled water to the ref-erence concentration of 0.06 microgramof candicidin activity per milliliter (es-timated).

(2) Loss on drying. Proceed as directedin § 436.200(b) of this chapter.

(3) pH. Proceed as directed in § 436.202of this chapter, using a 1 percent aque-ous suspension.

(4) Identity—(i) Preparation of aqueousalcohol solution. Prepare an aqueous al-cohol solution by mixing 53 volumes ofethyl alcohol and 47 volumes of water.

(ii) Preparation of standard solution.Grind a small portion of the candicidinworking standard to a fine powder witha mortar and pestle. Accurately weighan amount equivalent to 20,000micrograms of candicidin activity andtransfer it to a 100-milliliter volu-metric flask. Add about 50 millilitersof the aqueous alcohol solution andshake to effect complete dissolution.Bring to volume with the aqueous alco-hol solution and mix well. Transfer a25-milliliter aliquot to a 100-millilitervolumetric flask and bring to volumewith the aqueous alcohol solution. Thissolution contains 50 micrograms ofcandicidin activity per milliliter.

(iii) Preparation of sample solution.Proceed as directed in paragraph(b)(4)(ii) of this section.

(iv) Procedure. Using a suitable re-cording spectrophotometer, record theabsorption spectra of the standard so-lution and the sample solution betweenthe wavelengths of 330 and 410nanometers with the aqueous alcoholsolution as the reference solution.Compare the absorption spectra of thestandard solution and the sample solu-tion. They should exhibit absorptionmaxima and minima at the same wave-lengths, which are approximately 342,359, 378, and 397 nanometers for themaxima and 348, 366, and 390nanometers for the minima.

[39 FR 19134, May 30, 1974, as amended at 44FR 30333, May 25, 1979; 49 FR 2243, Jan. 19,1984]

§ 449.20 Griseofulvin.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Griseofulvin is a microsize,white to pale-cream compound withthe following chemical name: 7-chloro-2′,4,6-trimethoxy-6′β-methylspiro[benzofuran-2(3H),1′-[2]cyclohexene]-3,4′-dione. It is so puri-fied and dried that:

(i) Its griseofulvin content is not lessthan 900 micrograms and not morethan 1,050 micrograms of griseofulvinper milligram.

(ii) [Reserved](iii) Its loss on drying is not more

than 1.0 percent.(iv) Its melting point, after drying, is

not less than 217° C. and not more than224° C.

(v) Its specific rotation indimethylformamide at 25° C. is not lessthan ∂348° and not more than ∂364°.

(vi) Its ultraviolet absorption spec-trum in methyl alcohol comparesqualitatively with that of the griseo-fulvin reference standard.

(vii) Its residue on ignition is notmore than 0.2 percent.

(viii) Its heavy metals content is notmore than 25 parts per million.

(ix) Its specific surface area is notless than 1.3 and not more than 1.7square meters per gram.

(x) It is crystalline.


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(i) Results of tests and assays on thebatch for griseofulvin content, loss ondrying, melting point, specific rota-tion, identity, residue on ignition,heavy metals, specific surface area,and crystallinity.

(ii) Samples required: 10 packages,each containing not less than 1 gram.

(b) Tests and methods of assay—(1)Griseofulvin content (gas liquid chroma-tography). Proceed as directed in§ 436.321 of this chapter.


in § 436.200(b) of this chapter.(4) Melting point. Proceed as directed

in § 436.209 of this chapter.(5) Specific rotation. Accurately weigh

approximately 250 milligrams of thesample in a 25-milliliter glass-stoppered volumetric flask and dissolvein about 15 milliliters ofdimethylformamide. Bring to volumewith dimethylformamide, stopper, andmix well. Proceed as directed in§ 436.210 of this chapter, using a 2.0-dec-imeter polarimeter tube.

(6) Identity. Dissolve an accuratelyweighed portion of the sample and ofthe griseofulvin working standard anddissolve each in sufficient methyl alco-

hol to obtain a concentration of 10micrograms of griseofulvin per milli-liter and mix well. (The standard solu-tion can be kept under refrigerationand used for up to 1 month.) Record theultraviolet absorption spectrum of so-lutions of the sample and standardfrom 240 to 320 nanometers. The spec-tral curves shall be similar, and eachshall have a maximum at 292± 2nanometers and a minimum at 269± 2nanometers.

(7) Residue on ignition. Proceed as di-rected in § 436.207 of this chapter.

(8) Heavy metals. Proceed as directedin § 436.208 of this chapter.

(9) Specific surface area—(i) Procedure.Determine the apparent particle size inmicrons by the air-permeation method,using a suitable subsieve sizer. Weigh1.819 grams ±0.001 gram of the sampleand transfer to the compression tube ofthe apparatus. Compact the samplewith moderate pressure so that it has auniform porosity. Pass compressed dryair through the sample and measurethe air pressure with a water manom-eter. Observe the porosity and cal-culate the apparent particle size fromthe instrument equation or read itfrom a chart that has been calculatedin accordance with the equation. Re-peat the readings at successively high-er degrees of compaction until the ap-parent particle size reaches a mini-mum. Calculate the observed specificsurface area (SSA) in square meters pergram of sample, as follows:

Observed SSA =6

Minimum apparent particle sizein microns 1.455× × F

where F is a factor used to correct the appar-ent particle size to the true particle size:

Porosity reading F

0.80 ............................................................................ 1.37710.76 ............................................................................ 1.41420.72 ............................................................................ 1.45730.68 ............................................................................ 1.50820.64 ............................................................................ 1.56900.60 ............................................................................ 1.64320.56 ............................................................................ 1.73530.52 ............................................................................ 1.85280.48 ............................................................................ 2.00760.44 ............................................................................ 2.2203

Porosity reading F

0.40 ............................................................................ 2.5298

(ii) Standard. Determine the observedspecific surface area of the griseofulvinspecific surface area standard by themethod prescribed in paragraph(b)(9)(i) of this section, using the sameinstrument and the same air pressuresetting.


870

21 CFR Ch. I (4–1–98 Edition)§ 449.40

(iii) Calculations. Calculate the cor-rected specific surface area of the sam-ple as follows:

SSA of samassigned S

ple =Observed SSA of sample SA of standard

Observed SSA of standard

×

(10) Crystallinity. Proceed as directedin § 436.203(a) of this chapter.

[39 FR 19134, May 30, 1974, as amended at 44FR 20660, Apr. 6, 1979; 50 FR 19920, May 13,1985]

§ 449.40 Natamycin.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Natamycin is 22-[(3 - amino- 3,6 - dideoxy -β-D-mannopyranosyl) -oxy] - 1,3,26-trihydroxy - 12-methyl-10-oxo-6,11,28-trioxatricyclo [22.3.1.05,7] octacosa-8,14,16,18,20 - pentaene-25-carboxylicacid. It is an off-white to cream coloredpowder which may contain up to 3moles of water. It is practically insolu-ble in water, slightly soluble in meth-anol, and soluble in glacial acetic acidand dimethylformamide. It is so puri-fied and dried that:

(i) Its potency is not less than 900micrograms of natamycin per milli-gram on an anhydrous basis.

(ii) Its moisture content is not lessthan 6.0 percent and not more than 9.0percent.

(iii) Its pH in a 1 percent aqueous sus-pension is not less than 5.0 and notmore than 7.5.

(iv) It passes the identity test.(v) It is crystalline.(2) Labeling. It shall be labeled in ac-

cordance with the requirements of§ 432.5 of this chapter.

(3) Requests for certification; samples.In addition to complying with the re-quirements of § 431.1 of this chapter,each such request shall contain:

(i) Results of tests and assays on thebatch for potency, moisture, pH, iden-tity, and crystallinity.

(ii) Samples required: 10 packages,each containing approximately 500 mil-ligrams.

(b) Tests and methods of assay. Dilutesolutions of natamycin are very sen-

sitive to light and should be kept inthe dark as much as possible or sub-stantial decomposition will take place.

(1) Potency. Proceed as directed in§ 436.105 of this chapter, preparing thesample for assay as follows: Dissolve anaccurately weighed sample indimethylsulfoxide and further dilutewith sufficient dimethylsulfoxide togive a concentration of 100 microgramsof natamycin per milliliter (esti-mated). Further dilute with 0.2M potas-sium phosphate buffer, pH 10.5 (solu-tion 10), to the reference concentrationof 5.0 micrograms of natamycin permilliliter (estimated).

(2) Moisture. Proceed as directed in§ 436.201 of this chapter.

(3) pH. Proceed as directed in § 436.202of this chapter, using a 1.0 percentaqueous suspension.

(4) Identity. Accurately weigh ap-proximately 50 milligrams of the sam-ple into a 200-milliliter volumetricflask. Add approximately 5.0 millilitersof distilled water, and completelymoisten the sample. Then add approxi-mately 100 milliliters of an acid-alco-hol solvent (0.1 percent glacial aceticacid in methyl alcohol) and stir orshake mechanically in the dark untilsolution is complete. Dilute to volumewith the acid-alcohol solvent. Transfer2.0 milliliters of this solution to a 100-milliliter volumetric flask and diluteto volume with the acid-alcohol sol-vent. Using a suitable spectrophotom-eter with 1-centimeter cells and theacid-alcohol solvent as a blank, recordthe ultraviolet absorption spectrumfrom 215 to 330 nanometers. The spec-trum compares qualitatively to that ofthe natamycin working standard simi-larly treated.


[43 FR 55384, Nov. 28, 1978, as amended at 46FR 16684, Mar. 13, 1981]


871


§ 449.50 Nystatin.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin is the yellow tolight-tan compound of a kind of nys-tatin or a mixture of two or more suchcompounds. It is very slightly solublein water, moderately soluble in methylalcohol, butyl alcohol, or propyl alco-hol. It is so purified and dried that:

(i) Its potency is not less than 4,400units of nystatin per milligram; except,if it is packaged for extemporaneouspreparation of oral suspensions, its po-tency is not less than 5,000 units of nys-tatin per milligram.

(ii) [Reserved](iii) Its loss on drying is not more

than 5.0 percent.(iv) Its pH in a 3 percent aqueous sus-

pension is not less than 6.5 and notmore than 8.0.

(v) It passes the identity test.(vi) If it is packaged for extempo-

raneous preparation of oral suspen-sions, it passes the suspendibility test.

(vii) If it is packaged for extempo-raneous preparation of oral suspen-sions, it is crystalline.



(i) Results of tests and assays on thebatch for potency, loss on drying, pH,and identity. In addition, if it is pack-aged for extemporaneous preparationof oral suspensions, results of tests andassays on the batch for suspendibilityand crystallinity.

(ii) Samples required on the batch: 10packages, each containing approxi-mately 300 milligrams.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Dissolve an accu-rately weighed sample in sufficientdimethylformamide to give a nystatinconcentration of 400 units per milliliter(estimated). Further dilute with 10 per-cent potassium phosphate buffer, pH 6.0(solution 6), to the reference concentra-tion of 20 units of nystatin per milli-liter (estimated).

(2) [Reserved]


(4) pH. Proceed as directed in § 436.202of this chapter, using a 3 percent aque-ous suspension of the drug.

(5) Identity. Weigh approximately 100milligrams of the sample into a 200-milliliter, glass-stoppered, volumetricflask. Add 50 milliliters of absolutemethyl alcohol and 10 milliliters ofglacial acetic acid. When the samplehas dissolved, dilute to volume withmethyl alcohol. Transfer 2 millilitersof this solution to a 100-milliliter volu-metric flask and dilute to volume withmethyl alcohol. Use the same dilutionof acetic acid in methyl alcohol as theblank. Immediately determine the ab-sorption peaks at 230, 291, 305, and 319nanometers, and the shoulders at 279±2nanometers, using a suitable ultra-violet spectrophotometer and quartzcells. Set the instrument to 100 percenttransmission with the blank. If a re-cording spectrophotometer is used,record the ultraviolet absorption spec-trum from 220 nanometers to 350nanometers. If a nonrecording spectro-photometer is used, the exact positionsof the peaks and shoulder should be de-termined for the particular instrumentused. The ratio of the two absorbances

(A230/A279)

should be not less than 0.90 and notmore than 1.25.

(6) Suspendibility test. Transfer 200milligrams of the sample into a 250-milliliter beaker containing 200 milli-liters of water. Swirl the suspensiongently with a stirring rod. Allow thebeaker to remain still for 2 minutesand observe the bottom. It passes thetest if the powder remains in suspen-sion. If a significant amount of sedi-ment is observed, withdraw an accu-rately measured aliquot of the undis-turbed suspension and assay as di-rected in § 449.150c(b)(1) of this chapter.It passes the test if the suspension con-tains not less than 90 percent of thenumber of units of nystatin that it isrepresented to contain.


[39 FR 19134, May 30, 1974, as amended at 39FR 43833, Dec. 19, 1974; 49 FR 5098, Feb. 10,1984; 50 FR 19920, May 13, 1985]


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21 CFR Ch. I (4–1–98 Edition)§ 449.104

Subpart B—Oral Dosage Forms§ 449.104 Amphotericin B oral suspen-

sion.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Amphotericin B oral sus-pension is a mixture of amphotericin Bwith one or more suitable and harmlesspreservatives, colorings, sweetening in-gredients, flavorings, buffer sub-stances, lubricants, suspending agents,and sequestrants in an aqueous vehicle.Each milliliter contains 100 milligramsof amphotericin B. Its potency is satis-factory if it is not less than 90 percentand not more than 125 percent of thenumber of milligrams of amphotericinB that it is represented to contain. ItspH is not less than 4.5 and not morethan 6.0. The amphotericin B conformsto the standards prescribed by§ 449.4(a)(1).

(2) Labeling. It shall be labeled in ac-cordance with the requirements of§ 432.5 of this chapter.


(i) Results of tests and assays on:(a) The amphotericin B used in mak-

ing the batch for potency,amphotericin A content, loss on dry-ing, pH, residue on ignition, and iden-tity.

(b) The batch for potency and pH.(ii) Samples required:(a) The amphotericin B used in mak-

ing the batch: 10 packages, each con-taining approximately 500 milligrams.

(b) The batch: A minimum of 5 imme-diate containers.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Place an accuratelymeasured representative portion into ahigh-speed glass blender with sufficientdimethylsulfoxide to give a stock solu-tion of convenient concentration.Blend for 3 to 5 minutes. Dilute an ali-quot of the stock solution withdimethylsulfoxide to give a concentra-tion of 20 micrograms of amphotericinB per milliliter (estimated). Furtherdilute an aliquot with 0.2M potassiumphosphate buffer, pH 10.5 (solution 10),to the reference concentration of 1.0

microgram of amphotericin B per mil-liliter (estimated).

(2) pH. Proceed as directed in § 436.202of this chapter, using the undilutedsuspension.

[39 FR 19134, May 30, 1974, as amended at 50FR 19920, May 13, 1985]

§ 449.120 Griseofulvin oral dosageforms.

§ 449.120a Griseofulvin tablets.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Griseofulvin tablets are tab-lets composed of griseofulvin, with orwithout one or more suitable fillers,colorings, lubricants, and binders. Eachtablet contains 125, 250, or 500 milli-grams of griseofulvin. The griseofulvincontent is satisfactory if it is not lessthan 90 percent and not more than 115percent of the number of milligrams ofgriseofulvin that it is represented tocontain. The loss on drying is not morethan 5.0 percent. The tablets shall dis-integrate within 1 hour. The griseo-fulvin used conforms to the standardsprescribed by § 449.20(a)(1).



(i) Results of tests and assays on:(a) The griseofulvin used in making

the batch for griseofulvin content, losson drying, melting point, specific rota-tion, identity, residue on ignition,heavy metals, specific surface area,and crystallinity.

(b) The batch for griseofulvin con-tent, loss on drying, and disintegrationtime.

(ii) Samples required:(a) The griseofulvin used in making

the batch: 10 packages, each containingnot less than 1 gram.

(b) The batch for griseofulvin con-tent, loss on drying, and disintegrationtime.

(b) Tests and methods of assay—(1)Griseofulvin content (gas liquid chroma-tography). Proceed as directed in§ 436.321 of this chapter, except:

(i) Prepare the sample solution asfollows: Accurately weigh 20 tablets


873

Food and Drug Administration, HHS § 449.120b

and determine the average tabletweight. Grind the tablets to a fine pow-der in a mortar and transfer an accu-rately weighed sample to a volumetricflask of such size that for each 50 milli-liters of volume there are 40 milli-grams of griseofulvin (estimated). Addchloroform to about one-fourth volumeof the flask. Swirl the flask and applygentle heat to aid in dissolution of thegriseofulvin. Allow the mixture to cooland then dilute to volume with chloro-form and mix. Allow to settle andtransfer 2.0 milliliters of the supernateto a conical centrifuge tube and evapo-rate to dryness under a current of dryair. Add 1.0 milliliter of the internalstandard solution to the centrifugetube and mix vigorously to obtain auniform solution; and,

(ii) Calculate the milligrams of gris-eofulvin per tablet as follows:

Milligrams of griseofulvin

per tablet=

× × × ×

× × ×

R W f W V

R W

u s a u

s u 1,000 50where:

Ru=Area of the griseofulvin sample peak(at a retention time equal to that ob-served for the griseofulvin standard)/Area of the internal standard peak;

Rs=Area of the griseofulvin working stand-ard peak/Area of the internal standardpeak;

f=Potency of the griseofulvin workingstandard in micrograms per milligram;

Wa=Average tablet weight in milligrams;Ws=Weight of the griseofulvin working

standard in milligrams;Wu=Weight of the ground tablet powder

sample in milligrams;Vu=Volume of the dissolved ground tablet

powder sample in milliliters.


(3) Disintegration time. Proceed as di-rected in § 436.212 of this chapter, usingthe procedure described in paragraph(e)(1) of that section.


§ 449.120b Griseofulvin capsules.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Griseofulvin capsules aregelatin capsules containing griseo-fulvin with a suitable filler and binder,with or without a suitable lubricant.

Each capsule contains 125 or 250 milli-grams of griseofulvin. The griseofulvincontent is satisfactory if it is not lessthan 90 percent and not more than 115percent of the number of milligrams ofgriseofulvin that it is represented tocontain. The loss on drying is not morethan 1.0 percent. The griseofulvin usedconforms to the standards prescribedby § 449.20(a)(1).




the batch for griseofulvin content, losson drying, melting point, specific rota-tion, identity, residue on ignition,heavy metals, specific surface area,and crystallinity.

(b) The batch for griseofulvin contentand loss on drying.



(b) The batch: A minimum of 30 cap-sules.


(i) Prepare the sample solution asfollows: Empty the contents of 20 cap-sules into a tared weighing bottle.Weigh the powder and calculate the av-erage capsule weight. Mix the powderand transfer an accurately weighedsample to a volumetric flask of suchsize that for each 50 milliliters of vol-ume there are 40 milligrams of griseo-fulvin (estimated). Add chloroform toabout one-fourth volume of the flask.Swirl the flask and apply gentle heatto aid in dissolution of the griseo-fulvin. Allow the mixture to cool andthen dilute to volume with chloroformand mix. Allow to settle and transfer2.0 milliliters of the supernate to a con-ical centrifuge tube and evaporate todryness under a current of dry air. Add1.0 milliliter of the internal standardsolution to the centrifuge tube and mixvigorously to obtain a uniform solu-tion; and,


874

21 CFR Ch. I (4–1–98 Edition)§ 449.120c

(ii) Calculate the milligrams of gris-eofulvin per capsule as follows:

Milligrams of griseofulvinper capsule

=× × × ×

× × ×

R W f W V

R W

u s a u

s u 1,000 50where:



Ws=Weight of the griseofulvin workingstandard in milligrams;


Wa=Average capsule fill weight in milli-grams;

Wu=Weight of the ground tablet powdersample in milligrams;

Vu=Volume of the dissolved capsule powdersample in milliliters.

(2) Loss of drying. Proceed as directedin § 436.200(b) of this chapter.

[39 FR 19134, May 30, 1974, as amended at 43FR 9800, Mar. 10, 1978; 44 FR 20661, Apr. 6,1979; 50 FR 19920, May 13, 1985]

§ 449.120c Griseofulvin oral suspen-sion.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Griseofulvin oral suspensionis griseofulvin oral suspension with oneor more suitable flavorings, colorings,wetting agents, preservatives, anddiluents in an aqueous vehicle. Eachmilliliter contains 25 milligrams ofgriseofulvin. Its griseofulvin content issatisfactory if it is not less than 90 per-cent and not more than 115 percent ofthe number of milligrams of griseo-fulvin that it is represented to contain.Its pH is not less than 6.5 and not morethan 7.5. The griseofulvin used con-forms to the standards prescribed by§ 449.20(a)(1).

(2) Labeling. It shall be labeled in ac-cordance with the requirements of§ 432.5.



the batch for griseofulvin content, losson drying, melting point, specific rota-

tion, identity, residue on ignition,heavy metals, specific surface area,and crystallinity.

(b) The batch for griseofulvin contentand pH.





(i) Prepare the sample solution asfollows: Transfer an accurately meas-ured portion of the oral suspensionequivalent to 100 milligrams of griseo-fulvin into a 50-milliliter round-bot-tomed glass-stoppered centrifuge tube.Add 5 milliliters of water and 20 milli-liters of a solvent mixture of ethyl ace-tate and chloroform (85:15). Shake thetube for 1 minute and centrifuge itbriefly to separate the layers. Transfermost of the upper layer to a 100-milli-liter volumetric flask being careful notto remove any of the lower aqueouslayer. Repeat the extraction step withtwo additional 20-milliliter portions ofthe solvent mixture combining the ex-tracts in the volumetric flask with thefirst 20-milliliter extract. Dilute to vol-ume with the solvent mixture and mix.Place 2.0 milliliters of this solution ina conical centrifuge tube and evaporatethe contents to dryness on a steambath under a current of dry air. Add 1.0milliliter of the internal standard solu-tion to the centrifuge tube and mixvigorously to obtain a uniform solu-tion; and,

(ii) Calculate the milligrams of gris-eofulvin per milliliter as follows:

Milligrams of griseofulvinper milliliter

=× × ×

× ×

R W f 2

R V

u s

s o1,000where:





875

Food and Drug Administration, HHS § 449.120d


Vo=Volume of oral suspension taken inmilliliters.

(2) pH. Proceed as directed in § 436.202of this subchapter, using the undilutedsuspension.


§ 449.120d Griseofulvin(ultramicrosize) tablets.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Griseofulvin(ultramicrosize) tablets are composedof ultramicrosize crystals of griseo-fulvin which may or may not be dis-persed in polyethylene glycol 6,000.Each tablet contains 125, 165, 250, or 330milligrams of griseofulvin. The griseo-fulvin content is satisfactory if it isnot less than 90 percent and not morethan 115 percent of the number of milli-grams of griseofulvin that it is rep-resented to contain. The loss on dryingis not more than 5.0 percent. It passesthe solubility characteristic test. If itis dispersed in polyethylene glycol6,000, the griseofulvin used conforms tothe standards prescribed by§ 449.20(a)(1). If it is not dispersed inpolyethylene glycol 6,000, the griseo-fulvin used conforms to the standardsprescribed by § 449.20(a)(1), except spe-cific surface area.




the batch for potency, loss on drying,melting point, specific rotation, iden-tity, residue on ignition, heavy metals,specific surface area (if it is dispersedin polyethylene glycol 6,000), and crys-tallinity.

(b) The batch for griseofulvin con-tent, loss on drying, and solubilitycharacteristic.



(b) The batch: A minimum of 36 tab-lets.


(i) Prepare the sample solution asfollows: Accurately weigh 20 tabletsand determine the average tabletweight. Grind the tablets to a fine pow-der in a mortar and transfer an accu-rately weighed sample to a volumetricflask of such size that for each 50 milli-liters of volume there are 40 milli-grams of griseofulvin (estimated). Addchloroform to about one-fourth volumeof the flask. Swirl the flask and applygentle heat to aid in dissolution of thegriseofulvin. Allow the mixture to cooland then dilute to volume with chloro-form. Mix and allow to settle. Usinggentle vacuum, remove and discard thewaxy substance that forms on the topof the chloroform. Transfer 2.0 milli-liters of the chloroform solution to aconical centrifuge tube and evaporateto dryness under a current of dry air.Add 1.0 milliliter of the internal stand-ard solution to the centrifuge tube andmix vigorously to obtain a uniform so-lution; and,

(ii) Calculate the milligrams of gris-eofulvin per tablet as follows:

Milligrams ofgriseofulvin

per tablet=

× × × ×

× × ×

R W f W V

R W

u s a u

s u 1 000 50,where:





Wa=Average tablet weight in milligrams;Wu=Weight of the ground tablet powder

sample in milligrams;Vu=Volume of the dissolved ground tablet

powder sample in milliliters.



876

21 CFR Ch. I (4–1–98 Edition)§ 449.150

(3) Solubility characteristic test. Pro-ceed as directed in § 436.317 of this chap-ter.

[40 FR 41523, Sept. 8, 1975, as amended at 43FR 22676, May 26, 1978; 46 FR 7275, Jan. 23,1981; 46 FR 21361, Apr. 10, 1981; 46 FR 46313,Sept. 18, 1981; 47 FR 34132, Aug. 6, 1982; 50 FR19920, May 13, 1985]

§ 449.150 Nystatin oral dosage forms.

§ 449.150a Nystatin tablets.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin tablets are tabletscomposed of nystatin and suitable andharmless buffer substances, diluents,binders, lubricants, colorings, andflavorings. Each tablet contains 500,000units of nystatin. Its potency is satis-factory if it is not less than 90 percentand not more than 130 percent of thenumber of units of nystatin that it isrepresented to contain. The loss ondrying is not more than 8 percent. Thetablets shall disintegrate within 2hours. The nystatin used conforms tothe standards prescribed by§ 449.50(a)(1).



(i) Results of tests and assays on:(a) The nystatin used in making the

batch for potency, loss on drying, pH,and identity.

(b) The batch for potency, loss ondrying, and disintegration time.

(ii) Samples required:(a) The nystatin used in making the

batch: 10 packages, each consisting ofnot less than 300 milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Blend a representa-tive number of tablets for 3 to 5 min-utes in a high-speed glass blender withsufficient dimethylformamide to give aconvenient concentration. Dilute an al-iquot with sufficientdimethylformamide to give a stock so-lution containing 400 units of nystatin

per milliliter. Further dilute an ali-quot with 10 percent potassium phos-phate buffer, pH 6.0 (solution 6), to thereference concentration of 20 units ofnystatin per milliliter (estimated).


(3) Disintegration time. Proceed as di-rected in § 436.212 of this chapter.

[39 FR 19134, May 30, 1974, as amended at 50FR 19920, May 13, 1985; 50 FR 52772, Dec. 26,1985]

§ 449.150b Nystatin oral suspension.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin oral suspension isa suspension containing nystatin andone or more suitable preservatives, sus-pending agents, surfactants, flavorings,and colorings in purified water. Eachmilliliter contains 100,000 units of nys-tatin. Its potency is satisfactory if it isnot less than 90 percent and not morethan 130 percent of the number of unitsof nystatin that it is represented tocontain. Its pH is not less than 4.5 andnot more than 6.0; except, if the prod-uct contains glycerin, its pH is not lessthan 6.0 and not more than 7.5. Thenystatin used conforms to the stand-ards prescribed by § 449.50(a)(1).





(b) The batch for potency and pH.(ii) Samples required:(a) The nystatin used in making the

batch: 10 packages, each containing ap-proximately 300 milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Place an accuratelymeasured representative aliquot of thesample into a high-speed glass blenderjar containing sufficientdimethylformamide to give a conven-ient concentration. Blend for 3 to 5


877

Food and Drug Administration, HHS § 449.150d

minutes. Dilute an aliquot with suffi-cient dimethylformamide to give astock solution containing 400 units ofnystatin per milliliter (estimated). Re-move and dilute with 10 percent potas-sium phosphate buffer, pH 6.0 (solution6), to the reference concentration of 20units of nystatin per milliliter (esti-mated).



§ 449.150c Nystatin for oral suspen-sion.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Nystatin for oral suspensionis a dry powder consisting of nystatin,and suitable and harmless suspendingsubstances, preservatives, diluents,colorings, and flavorings. When thesuspension is prepared as directed inits labeling, each milliliter contains100,000 units of nystatin. Its potency issatisfactory if it is not less than 90 per-cent and not more than 140 percent ofaliquot of the stock solution and fur-ther the number of units of nystatinthat it is represented to contain. ThepH of the reconstituted drug is not lessthan 4.9 and not more than 5.5. Itsmoisture content is not more than 7.0percent. The nystatin used conforms tothe standards prescribed by§ 449.50(a)(1).





(b) The batch for potency, moistureand pH.


batch: 10 packages, each consisting of300 milligrams.

(b) The batch: A minimum of five im-mediate containers.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 of

this chapter, preparing the sample forassay as follows: Reconstitute the drugas directed in the labeling. Blend anappropriate aliquot in a high-speedglass blender for 3 to 5 minutes, usingsufficient dimethylformamide to give aconvenient concentration. Dilute an al-iquot with sufficientdimethylformamide to give a stock so-lution containing 400 units of nystatinper milliliter. Further dilute an ali-quot with 10 percent potassium phos-phate buffer, pH 6.0 (solution 6), to thereference concentration of 20 units ofnystatin per milliliter (estimated).

(2) Moisture. Using the dry powder,proceed as directed in § 436.201 of thischapter.

(3) pH. Proceed as directed in § 436.202of this chapter, using the suspensionafter reconstituting as directed in thelabeling.


§ 449.150d Nystatin pastilles.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin pastilles are com-posed of nystatin with suitablediluents, binders, buffers, colorings,and flavorings. Each pastille containsnystatin equivalent to 200,000 units ofnystatin. Its potency is satisfactory ifit contains not less than 90 percent andnot more than 125 percent of the num-ber of units of nystatin that it is rep-resented to contain. The pH in an aque-ous solution is not less than 5.0 and notmore than 7.5. It disintegrates within90 minutes. The nystatin used con-forms to the standards prescribed by§ 449.50(a)(1).





(b) The batch for potency, pH, anddisintegration time.

(ii) Samples, if required by the Direc-tor, Center for Drug Evaluation andResearch:


878

21 CFR Ch. I (4–1–98 Edition)§ 449.204

(a) The nystatin used in making thebatch: 10 packages, each containing ap-proximately 300 milligrams.

(b) The batch: A minimum of 36 pas-tilles.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Place a representativenumber of pastilles into a high-speedglass blender jar containing 100 milli-liters of sterile distilled water. Blendfor 18 to 20 minutes. Add 400 millilitersof dimethylformamide and continueblending for an additional 10 minutes.Remove an aliquot and add sufficient80 percent dimethylformamide so thatupon final dilution with 10 percent po-tassium phosphate buffer, pH 6.0 (solu-tion 6), to the reference concentrationof 20 units of nystatin per milliliter,the concentration ofdimethylformamide will be 4 percent.

(2) pH. Dissolve 1 pastille in 100 milli-liters of distilled water at 37 °C, cool,and proceed as directed in § 436.202 ofthis chapter.

(3) Disintegration time. Proceed as di-rected in § 436.212 of this chapter, usingthe method described in paragraph(e)(4) of that section.

[52 FR 4617, Feb. 13, 1987; 52 FR 7741, Mar. 12,1987, as amended at 55 FR 11584, Mar. 29, 1990]

Subpart C—Injectable DosageForms

§ 449.204 Amphotericin B for injection.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Amphotericin B for injec-tion is a dry mixture containing ineach immediate container 50 milli-grams of amphotericin B, 41 milligramsof sodium desoxycholate, and suitablebuffering substances. Its potency issatisfactory if it is not less than 90 per-cent and not more than 120 percent ofthe number of milligrams ofamphotericin B that it is representedto contain. It is sterile. It isnonpyrogenic. Its loss on drying is notmore than 8.0 percent. Its pH in anaqueous solution containing 10 milli-grams of amphotericin B per milliliteris not less than 7.2 and not more than8.0. The amphotericin B used conforms

to the standards prescribed by§ 449.4a(a)(1).

(2) Labeling. In addition to the label-ing requirements prescribed by § 432.5of this chapter, each package shall bearon its label and labeling the followingstatement: ‘‘For intravenous infusionin hospitals only’’.



ing the batch for potency,amphotericin A content, loss on dry-ing, pH, residue on ignition, and iden-tity.

(b) The batch for potency, sterility,pyrogens, loss on drying, and pH.

(ii) Samples required:(a) Amphotericin B used in making

the batch: 10 packages, each containingapproximately equal portions of notless than 500 milligrams.

(b) The batch:(1) For all tests except sterility: A

minimum of 10 immediate containers.(2) For sterility testing: 20 immediate

containers collected at regular inter-vals throughout each filling operation.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Reconstitute as di-rected in the labeling. Then using asuitable syringe and hypodermic nee-dle, remove all of the withdrawablecontents if the container is representedas a single-dose container; or, if the la-beling specifies the amount of potencyin a given volume of the resultantpreparation, remove an accuratelymeasured representative portion fromeach container. Dilute with sufficientdimethylsulfoxide to give a stock solu-tion of convenient concentration. Fur-ther dilute an aliquot of the stock solu-tion with dimethylsulfoxide to a con-centration of 20 micrograms ofamphotericin B per milliliter (esti-mated). Remove an aliquot of this solu-tion and dilute with 0.2M potassiumphosphate buffer, pH 10.5 (solution 10),to the reference concentration of 1.0microgram of amphotericin B per mil-liliter (estimated).


879


(2) Sterility. Proceed as directed in§ 436.20 of this chapter, using the meth-od described in paragraph (e)(1) of thatsection, except use 50 milligrams inlieu of 300 milligrams.

(3) [Reserved](4) Pyrogens. Proceed as directed in

§ 436.32(e) of this chapter, using a solu-tion containing 2 milligrams ofamphotericin B per milliliter, except inlieu of paragraph (a)(3), if no rabbitshows an individual rise in tempera-ture of 1.1° C. or more above its respec-tive control temperature, and if thesum of the three temperature risesdoes not exceed 3° C., the sample meetsthe requirements for absence ofpyrogen. If one or two rabbits show atemperature rise of 1.1° C. or more, orif the sum of temperature rises exceeds3° C., repeat the test using five otherrabbits. If not more than three of theeight rabbits show a temperature riseof 1.1° C. or more, and if the sum of thetemperature rises does not exceed 8° C.the sample meets the requirements forabsence of pyrogens.


(6) pH. Proceed as directed in § 436.202of this chapter using an aqueous solu-tion containing 10 milligrams ofamphotericin B per milliliter.

[39 FR 19134, May 30, 1974, as amended at 45FR 16472, Mar. 14, 1980; 50 FR 19920, May 13,1985]

Subpart D—Ophthalmic DosageForms

§ 449.340 Natamycin ophthalmic sus-pension.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Natamycin ophthalmic sus-pension contains natamycin with oneor more suitable and harmless preserv-atives in a suitable and harmless aque-ous vehicle. Each milliliter contains 50milligrams of natamycin. Its potencyis satisfactory if it is not less than 90percent and not more than 125 percentof the number of milligrams ofnatamycin that it is represented tocontain. It is sterile. Its pH is not lessthan 5.0 and not more than 7.5. Thenatamycin used conforms to the stand-ards prescribed by § 449.40(a)(1).



(i) Results of tests and assays on:(a) The natamycin used in making

the batch for potency, moisture, pH,identity, and crystallinity.

(b) The batch for potency, sterility,and pH.

(ii) Samples required:(a) The natamycin used in making

the batch: 10 packages, each containingnot less than 500 milligrams.

(b) The batch:(1) For all tests except sterility: A

minimum of five immediate contain-ers.

(2) For sterility testing: 20 immediatecontainers, collected at regular inter-vals throughout each filling operation.

(b) Tests and methods of assay. Dilutesolutions of natamycin are very sen-sitive to light and should be kept inthe dark as much as possible or sub-stantial decomposition will take place.

(1) Potency. Proceed as directed in§ 436.105 of this chapter, preparing thesample for assay as follows: Dilute anaccurately measured representativeportion of the sample with sufficientdimethylsulfoxide to give a stock solu-tion of convenient concentration. Fur-ther dilute an aliquot of the stock solu-tion with dimethylsulfoxide to a con-centration of 100 micrograms ofnatamycin per milliliter (estimated).Further dilute an aliquot with 0.2M po-tassium phosphate buffer, pH 10.5 (solu-tion 10), to the reference concentrationof 5.0 micrograms of natamycin permilliliter (estimated).

(2) Sterility. Proceed as directed in§ 436.20 of this chapter, using the meth-od described in paragraph (e)(2) of thatsection, except use 0.25 milliliter ofsample in lieu of 1.0 milliliter.


[43 FR 55384, Nov. 28, 1978, as amended at 48FR 51293, Nov. 8, 1983]


880

21 CFR Ch. I (4–1–98 Edition)§ 449.504

Subpart E [Reserved]

Subpart F—Dermatologic DosageForms

§ 449.504 Amphotericin B dermatologicdosage forms.

§ 449.504a Amphotericin B ointment.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Amphotericin B ointment iscomposed of amphotericin B in a suit-able and harmless ointment base. Itmay contain suitable and harmlesscoloring agents and protectants. Itcontains 30 milligrams of amphotericinB in each gram. Its potency is satisfac-tory if it is not less than 90 percent andnot more than 125 percent of the num-ber of milligrams of amphotericin Bthat it is represented to contain. Itsmoisture content is not more than 1.0percent. The amphotericin B used con-forms to the standards prescribed by§ 449.4(a)(1) (i), (ii), (v), (vi), and (vii).




ing the batch for potency,amphotericin A content, pH, residue onignition, and identity.

(b) The batch for potency and mois-ture.

(ii) Samples required:(a) Amphotericin B used in making



(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Place an accuratelyweighed representative portion of thesample (usually 1 gram) into an appro-priate-sized Erlenmeyer flask with 10milliliters of ethyl ether. Allow to dis-solve for 1 hour with the intermittentmanual shaking. Add a measuredamount of dimethylsulfoxide to theflask and place on a shaker for 10 min-utes. Further dilute with

dimethylsulfoxide to a concentrationof 20 micrograms of amphotericin B permilliliter (estimated). Remove an ali-quot and dilute with 0.2M potassiumphosphate buffer, pH 10.5 (solution 10),to the reference concentration of 1.0microgram of amphotericin B per mil-liliter (estimated).


§ 449.504b Amphotericin B cream.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Amphotericin B cream iscomposed of amphotericin B, with orwithout one or more suitable andharmless emollients, perfumes,dispersants, and preservatives, in asuitable and harmless cream base. Itcontains 30 milligrams of amphotericinB in each gram. Its potency is satisfac-tory if it is not less than 90 percent andnot more than 125 percent of the num-ber of milligrams of amphotericin Bper gram that it is represented to con-tain. The amphotericin B used con-forms to the standards prescribed by§ 449.4(a)(1) (i), (ii), (v), (vi), and (vii).





(b) The batch for potency.(ii) Samples required:(a) Amphotericin B used in making



(b) Tests and methods of assay; po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: With the aid of ahigh-speed glass blender, dissolve anaccurately weighed sample in suffi-cient dimethylsulfoxide to give a stocksolution of convenient concentration.Further dilute with dimethylsulfoxideto a concentration of 20 micrograms of


881

Food and Drug Administration, HHS § 449.550a

amphotericin B per milliliter (esti-mated). Remove an aliquot and dilutewith 0.2M potassium phosphate buffer,pH 10.5 (solution 10), to the referenceconcentration of 1.0 microgram ofamphotericin B per milliliter (esti-mated).

§ 449.504c Amphotericin B lotion.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Amphotericin B lotion iscomposed of amphotericin B in a suit-able and harmless lotion vehicle. Itcontains suitable and harmlessemollients, emulsifiers, coloringagents, diluents, preservatives, andperfumes. It contains 30 milligrams ofamphotericin B per milliliter. Its po-tency is satisfactory if it is not lessthan 90 percent and not more than 125percent of the number of milligrams ofamphotericin B per milliliter that it isrepresented to contain. Its pH is notless than 5.0 and not more than 7.0. Theamphotericin B used conforms to thestandards prescribed by § 449.4(a)(1) (i),(ii), (v), (vi), and (vii).





(b) The batch for potency and pH.(ii) Samples required:(a) The amphotericin B used in mak-

ing the batch: 10 packages, each con-taining not less than 500 milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Dissolve an aliquot insufficient dimethylsulfoxide to give astock solution of convenient con-centration. Further dilute the stocksolution with dimethylsulfoxide to aconcentration of 20 micrograms ofamphotericin B per milliliter (esti-mated). Remove an aliquot and dilutewith 0.2M potassium phosphate buffer,

pH 10.5 (solution 10), to the referenceconcentration of 1.0 microgram ofamphotericin B per milliliter (esti-mated).

(2) pH. Proceed as directed in § 436.202of this chapter, using the undiluted lo-tion.

§ 449.550 Nystatin dermatologic dosageforms.

§ 449.550a Nystatin ointment.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin ointment is com-posed of nystatin and a suitable andharmless ointment base. Each gramcontains 100,000 units of nystatin. Itspotency is satisfactory if it is not lessthan 90 percent and not more than 130percent of the number of units of nys-tatin that it is represented to contain.The moisture content is not more than0.5 percent. The nystatin used con-forms to the standards prescribed by§ 449.50(a)(1) (i), (iii), (iv), and (v).







batch: 10 containers, each consisting of300 milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Using sufficientdimethylformamide to give a con-centration of 400 units of nystatin (es-timated) per milliliter, blend an accu-rately weighed representative portionin a high-speed glass blender for 3 to 5minutes. Further dilute with 10 percentpotassium phosphate buffer, pH 6 (solu-tion 6), to the reference concentrationof 20 units of nystatin per milliliter(estimated).


882

21 CFR Ch. I (4–1–98 Edition)§ 449.550b


§ 449.550b Nystatin-iodochlorhydroxyquin ointment.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Nystatin-iodochlorhydroxyquin ointment iscomposed of nystatin andiodochlorhydroxyquin in a suitable andharmless ointment base. Each gramcontains 100,000 units of nystatin and 10milligrams of iodochlorhydroxyquin.Its nystatin content is satisfactory if itis not less than 90 percent and notmore than 140 percent of the number ofunits of nystatin that it is representedto contain. Its iodochlorhydroxyquincontent is satisfactory if it is not lessthan 90 percent and not more than 110percent of the number of milligrams ofiodochlorhydroxyquin that it is rep-resented to contain. It passes the iden-tity test for iodochlorhydroxyquin. Itsmoisture content is not more than 0.5percent. The nystatin used conforms tothe standards prescribed by§ 449.50(a)(1). The iodochlorhydroxyquinused conforms to the standards pre-scribed by U.S.P. XVIII.





(b) The iodochlorhydroxyquin used inmaking the batch for all U.S.P. XVIIIspecifications.

(c) The batch for nystatin content,iodochlorhydroxyquin content,iodochlorhydroxyquin identity, andmoisture.



(b) The batch: A minimum of sevenimmediate containers.

(b) Tests and methods of assay—(1)Nystatin content. Proceed as directed in§ 436.105 of this chapter, preparing thesample for assay as follows: Place an

accurately weighed representative por-tion of the sample into a high-speedglass blender jar containing sufficientdimethylformamide to give a conven-ient concentration. Blend for 3 to 5minutes. Remove an aliquot and dilutewith sufficient dimethylformamide toyield a stock solution containing 400units of nystatin per milliliter (esti-mated). Further dilute an aliquot ofthe stock solution with 10 percent po-tassium phosphate buffer, pH 6.0 (solu-tion 6), to the reference concentrationof 20 units of nystatin per milliliter(estimated).

(2) Iodochlorhydroxyquin content—(i)Reagents. (a) Ferric chloride reagent.Dissolve 1.0 gram of ferric chloride(FeCl3·6H2O) in a mixture of 1.0 milli-liter of concentrated hydrochloric acidand sufficient distilled water to make 1liter.

(b) Acetone, reagent grade.(c) 2-Methoxyethanol, reagent grade.(ii) Preparation of standard solution.

Dissolve an accurately weighed portionof iodochlorhydroxyquin U.S.P. ref-erence standard in sufficient 2-methoxyethanol to make a solutioncontaining 1.0 milligram ofiodochlorhydroxyquin per milliliter.Transfer 5.0 milliliters of this standardsolution to a 50-milliliter volumetricflask.

(iii) Preparation of sample solution. Ac-curately weigh a portion of the sampleequivalent to 50 milligrams ofiodochlorhydroxyquin into a 125-milli-liter Erlenmeyer flask. Add 50 milli-liters of acetone, warm on a steambath, and shake gently. Cool to roomtemperature and filter contentsthrough a pledget of glass wool into a100-milliliter volumetric flask. Washthe Erlenmeyer flask with two 20-milli-liter portions of acetone and filter thewashings into the volumetric flask. Di-lute to volume with acetone and mixthoroughly. Transfer a 10-milliliter ali-quot of the acetone solution to a 50-milliliter volumetric flask and evapo-rate on a steam bath. To the residue,add 20 milliliters of 2-methoxyethanoland swirl to dissolve theiodochlorhydroxyquin.

(iv) Procedure. To each flask contain-ing standard solution and sample solu-tion, respectively, add 2.0 milliliters offerric chloride reagent and dilute to


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Food and Drug Administration, HHS § 449.550c

volume with 2-methoxyethanol. Mixthoroughly. Using a suitable spectro-photometer equipped with 1.0-centi-meter cells and a blank prepared by di-luting 2.0 milliliters of ferric chloridereagent to 50 milliliters with 2-methoxyethanol, determine the absorb-ance of the sample and standard solu-tions at 650 nanometers. Set the instru-ment to 100-percent transmission withthe blank.

(v) Calculation.

Milligrams of iodochlorhydroxyquin pergram of sample=(Absorbance of sample ×50)/(Absorbance of standard × Weight ofsample in grams)

(3) Iodochlorhydroxyquin identity. Pro-ceed as directed in § 436.400 of this chap-ter, preparing the sample solution asfollows: Accurately weigh a portion ofthe sample equivalent to 50 milligramsof iodochlorhydroxyquin into a 125-mil-liliter Erlenmeyer flask. Add 50 milli-liters of acetone, warm on a steambath, and shake gently. Cool to roomtemperature and filter contentsthrough a pledget of glass wool into a100-milliliter volumetric flask. Washthe Erlenmeyer flask with two 20-milli-liter portions of acetone and filter thewashings into the volumetric flask. Di-lute to volume with acetone and mixthoroughly.



§ 449.550c Nystatin-neomycin sulfate-gramicidin-triamcinolone acetonideointment; nystatin-neomycin sul-fate-gramicidin-fludrocortisone ace-tate ointment.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. The drug is nystatin, neo-mycin sulfate, gramicidin, and eithertriamcinolone acetonide orfludrocortisone acetate in a suitableointment base. Each gram contains100,000 units of nystatin, 2.5 milligramsof neomycin, 0.25 milligram of grami-cidin, and either 1.0 milligram oftriamcinolone acetonide or 1.0 milli-gram of fludrocortisone acetate. Itsnystatin content is satisfactory if it isnot less than 90 percent and not morethan 140 percent of the number of unitsof nystatin that it is represented to

contain. Its neomycin content is satis-factory if it is not less than 90 percentand not more than 140 percent of thenumber of milligrams of neomycin thatit is represented to contain. Its grami-cidin content is satisfactory if it is notless than 90 percent and not more than140 percent of the number of milli-grams of gramicidin that it is rep-resented to contain. Its moisture con-tent is not more than 0.5 percent. Thenystatin used conforms to the stand-ards prescribed by § 449.50(a)(1) (i), (iii),(iv), and (v). The neomycin sulfate usedconforms to the standards prescribedby § 444.42a(a)(1) of this chapter. Thegramicidin used conforms to the stand-ards prescribed by § 448.25(a)(1) (i), (iii),(iv), (v), and (vi) of this chapter.





(b) The neomycin sulfate used inmaking the batch for potency, loss ondrying, pH, and identity.

(c) The gramicidin used in makingthe batch for potency, loss on drying,residue on ignition, melting point,crystallinity, and identity.

(d) The batch for potency and mois-ture.



(b) The neomycin sulfate used inmaking the batch: 10 packages, eachconsisting of 300 milligrams.

(c) The gramicidin used in makingthe batch: 10 packages, each consistingof 500 milligrams.

(d) The batch: A minimum of sevenimmediate containers.

(b) Tests and methods of assay—(1) Po-tency—(i) Nystatin content. Proceed asdirected in § 436.105 of this chapter, pre-paring the sample for assay as follows:Blend an accurately weighed represent-ative portion in a high-speed glassblender for 3 to 5 minutes with suffi-cient dimethylformamide to give a


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21 CFR Ch. I (4–1–98 Edition)§ 449.550d

concentration of 400 units of nystatinper milliliter (estimated). Further di-lute with 10 percent potassium phos-phate buffer, pH 6.0 (solution 6), to thereference concentration of 20 units ofnystatin per milliliter (estimated).

(ii) Neomycin content. Proceed as di-rected in § 436.105 of this chapter, pre-paring the sample for assay as follows:Place an accurately weighed represent-ative portion of the ointment into aseparatory funnel containing 50 milli-liters of peroxide-free ether. Shake thesample and ether until homogenous.Add 20 to 25 milliliters of 0.1M potas-sium phosphate buffer, pH 8.0 (solution3), and shake well. Allow the layers toseparate. Remove the buffer layer andrepeat the extraction with new por-tions of the buffer at least three timesand any additional times necessary toinsure complete extraction of the anti-biotic. Combine the extractives and ad-just to an appropriate volume to give astock solution of convenient con-centration. Further dilute an aliquot ofthe stock solution with solution 3 tothe reference concentration of 1.0microgram of neomycin per milliliter(estimated).

(iii) Gramicidin content. Proceed as di-rected in §436.106 of this chapter, pre-paring the sample for assay as follows:Accurately weigh and dissolve a rep-resentative portion of the sample in ap-proximately 50 milliliters of petroleumether in a separatory funnel. Extractwith 20 milliliters of 80 percent alcoholprepared from alcohol U.S.P. XX. Re-peat the extraction three times. Com-bine the extractives in a suitable volu-metric flask, bring to volume with al-cohol U.S.P. XX, and mix well. Furtherdilute with alcohol U.S.P. XX to thereference concentration of 0.04microgram of gramicidin per milliliter(estimated).


[39 FR 19134, May 30, 1974, as amended at 47FR 23710, June 1, 1982; 50 FR 19920, May 13,1985]

§ 449.550d Nystatin cream.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin cream is composedof nystatin and suitable and harmlessemulsifiers, perfumes, buffers, preserv-

atives, and a protectant in a suitableand harmless cream base. Each gramcontains 100,000 units of nystatin. Itspotency is satisfactory if it is not lessthan 90 percent and not more than 130percent of the number of units of nys-tatin that it is represented to contain.The nystatin used conforms to thestandards prescribed by § 449.50(a)(1) (i),(iii), (iv), and (v).





(b) The batch for potency.(ii) Samples required:(a) The nystatin used in making the

batch: 10 containers, each consisting of300 milligrams.


(b) Tests and methods of assay; po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Using sufficientdimethylformamide to give an esti-mated concentration of 400 units ofnystatin per milliliter, blend an accu-rately weighed representative portionin a high-speed blender for 3 to 5 min-utes. Further dilute with 10 percent po-tassium phosphate buffer, pH 6.0 (solu-tion 6), to the reference concentrationof 20 units of nystatin per milliliter(estimated).

§ 449.550e Nystatin-neomycin sulfate-gramicidin-triamcinolone acetonidecream.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Nystatin-neomycin sulfate-gramicidin-triamcinolone acetonidecream is composed of nystatin, neomy-cin sulfate, gramicidin, triamcinoloneacetonide, and suitable and harmlessemulsifiers, solvents, perfumes, buff-ers, preservatives, and a protectant ina suitable cream base. Each gram con-tains 100,000 units of nystatin, 2.5 milli-grams of neomycin, 0.25 milligram ofgramicidin, and 1 milligram of


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Food and Drug Administration, HHS § 449.550f

triamcinolone acetonide. Its nystatincontent is satisfactory if it is not lessthan 90 percent and not more than 140percent of the number of units of nys-tatin that it is represented to contain.Its neomycin content is satisfactory ifit is not less than 90 percent and notmore than 140 percent of the number ofmilligrams of neomycin that it is rep-resented to contain. Its gramicidincontent is satisfactory if it is not lessthan 90 percent and not more than 140percent of the number of milligrams ofgramicidin that it is represented tocontain. The nystatin used conforms tothe standards prescribed by§ 449.50(a)(1) (i), (iii), (iv), and (v). Theneomycin sulfate used conforms to thestandards prescribed by § 444.42(a)(1) ofthis chapter. The gramicidin used con-forms to the standards prescribed by§ 448.25(a)(1) (i), (iii), (iv), (v), and (vi) ofthis chapter.







(d) The batch for nystatin content,neomycin content, and gramicidin con-tent.






(b) Tests and methods of assay—(1) Po-tency—(i) Nystatin content. Proceed asdirected in § 436.105 of this chapter, pre-

paring the sample for assay as follows:Using sufficient dimethylformamide togive a concentration of 400 units ofnystatin (estimated) per milliliter,blend an accurately weighed represent-ative portion in a high-speed glassblender for 3 to 5 minutes. Further di-lute with 10 percent potassium phos-phate buffer, pH 6.0 (solution 6), to thereference concentration of 20 units ofnystatin per milliliter (estimated).

(ii) Neomycin content. Proceed as di-rected in § 436.105 of this chapter, pre-paring the sample for assay as follows:Place an accurately weighed represent-ative portion of the cream in a sepa-ratory funnel containing 50 millilitersof peroxide-free ether. Shake the sam-ple and either until homogeneous. Add20 to 25 milliliters of 0.1M potassiumphosphate buffer, pH 8.0 (solution 3),and shake well. Allow the layers toseparate. Remove the buffer layer andrepeat the extraction with new por-tions of the buffer at least three timesand any additional times necessary toensure complete extraction of the anti-biotic. Combine the extractives and ad-just to an appropriate volume to give astock solution of convenient con-centration. Further dilute an aliquot ofthe stock solution with solution 3 tothe reference concentration of 1.0microgram of neomycin per milliliter(estimated).

(iii) Gramicidin content. Proceed as di-rected in §436.106 of this chapter, pre-paring the sample for assay as follows:Accurately weigh a representative por-tion of the sample and dissolve in ap-proximately 50 milliliters of petroleumether in a separatory funnel. Extractwith 20 milliliters of 80 percent alcoholprepared from alcohol U.S.P. XX. Re-peat the extraction three times. Com-bine the extractives in a suitable volu-metric flask, bring to volume with al-cohol U.S.P. XX, and mix well. Furtherdilute with alcohol U.S.P. XX to thereference concentration of 0.04microgram of gramicidin per milliliter(estimated).

[39 FR 19134, May 30, 1974, as amended at 47FR 23710, June 1, 1982; 50 FR 19920, May 13,1985]

§ 449.550f Nystatin topical powder.(a) Requirements for certification—(1)

Standards of identity, strength, quality,


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21 CFR Ch. I (4–1–98 Edition)§ 449.550g

and purity. Nystatin topical powder is adry powder composed of nystatin andtalc. Each gram contains 100,000 unitsof nystatin. Its potency is satisfactoryif it is not less than 90 percent and notmore than 130 percent of the number ofunits of nystatin that it is representedto contain. Its loss on drying is notmore than 2.0 percent. The nystatinused conforms to the standards pre-scribed by § 449.50(a)(1) (i), (iii), (iv),and (v).





(b) The batch for potency and loss ondrying.


batch: 10 packages, each containing 300milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Blend an accuratelyweighed representative sample for 3 to5 minutes in a high-speed glass blenderwith sufficient dimethylformamide togive a convenient concentration. Di-lute with sufficientdimethylformamide to yield a stock so-lution containing 400 units of nystatinper milliliter (estimated). Further di-lute with 10 percent potassium phos-phate buffer, pH 6.0 (solution 6), to thereference concentration of 20 units ofnystatin per milliliter (estimated).


§ 449.550g Nystatin-neomycin sulfate-gramicidin topical powder.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Nystatin-neomycin sulfate-gramicidin topical powder is a dry pow-der composed of nystatin, neomycinsulfate, gramicidin, and talc. Eachgram contains 100,000 units of nystatin,

2.5 milligrams of neomycin and 0.25milligram of gramicidin. Its nystatincontent is satisfactory if it is not lessthan 90 percent and not more than 140percent of the number of units of nys-tatin that it is represented to contain.Its neomycin content is satisfactory ifit is not less than 90 percent and notmore than 140 percent of the number ofmilligrams of neomycin that it is rep-resented to contain. Its gramicidincontent is satisfactory if it is not lessthan 90 percent and not more than 140percent of the number of milligrams ofgramicidin that it is represented tocontain. Its loss on drying is not morethan 2.0 percent. The nystatin usedconforms to the standards prescribedby § 449.50(a)(1) (i), (iii), (iv), and (v).The neomycin sulfate used conforms tothe standards prescribed by§ 444.42a(a)(1) of this chapter. The gram-icidin used conforms to the standardsprescribed by § 448.25(a)(1) (i), (iii), (iv),(v) and (vi) of this chapter.







(d) The batch for nystatin content,neomycin content, gramicidin content,and loss on drying.


batch: 10 packages each consisting of300 milligrams.





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(b) Tests and methods of assay—(1) Po-tency—(i) Nystatin content. Proceed asdirected in § 436.105 of this chapter, pre-paring the sample for assay as follows:Blend the entire contents of an accu-rately weighed representative portionof the sample for 3 to 5 minutes in ahigh-speed glass blender with sufficientdimethylformamide to give a conven-ient concentration. Dilute with suffi-cient dimethylformamide to yield astock solution containing 400 units ofnystatin per milliliter (estimated).Further dilute with 10 percent potas-sium phosphate buffer, pH 6.0 (solution6), to the reference concentration of 20units of nystatin per milliliter (esti-mated).

(ii) Neomycin content. Proceed as di-rected in § 436.105 of this chapter, pre-paring the sample for assay as follows:Blend an accurately weighed represent-ative sample for 3 to 5 minutes in suffi-cient 0.1M potassium phosphate buffer,pH 8 (solution 3), to give a convenientconcentration. Further dilute an ali-quot with solution 3 to the referenceconcentration of 1 microgram of neo-mycin per milliliter (estimated).

(iii) Gramicidin content. Proceed as di-rected in §436.106 of this chapter, pre-paring the sample for assay as follows:Dissolve an accurately weighed rep-resentative sample in alcohol U.S.P.XX and filter. Collect the filtrate anddilute a portion with alcohol U.S.P. XXto the reference concentration of 0.04microgram of gramicidin per milliliter(estimated).


[39 FR 19134, May 30, 1974, as amended at 41FR 10886, Mar. 15, 1976; 47 FR 23710, June 1,1982; 50 FR 19920, May 13, 1985]

§ 449.550h Nystatin lotion.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Nystatin lotion is composedof nystatin with one or more suitableand harmless suspending agents, emul-sifiers, surfactants, and preservativesin a suitable and harmless vehicle.Each milliliter contains 100,000 units ofnystatin. Its potency is satisfactory ifit is not less than 90 percent and notmore than 140 percent of the number ofunits of nystatin that it is representedto contain. Its pH is not less than 5.5

and not more than 7.5. The nystatinused conforms to the standards pre-scribed by § 449.50(a)(1).





(b) The batch for potency and pH.(ii) Samples required:(a) The nystatin used in making the



(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Place an accuratelymeasured representative portion of thesample into a high-speed glass blenderjar containing sufficientdimethylformamide to give a conven-ient concentration. Blend for 3 to 5minutes. Remove an aliquot and dilutewith sufficient dimethylformamide toyield a stock solution containing 400units of nystatin per milliliter (esti-mated). Further dilute with 10 percentpotassium phosphate buffer, pH 6.0 (so-lution 6), to the reference concentra-tion of 20 units of nystatin per milli-liter (estimated).

(2) pH. Proceed as directed in § 436.202of this chapter, using the undilutedsample.

[40 FR 3766, Jan. 24, 1975, as amended at 50FR 19920, May 13, 1985]

Subpart G—Vaginal DosageForms

§ 449.610 Candicidin vaginal dosageforms.

§ 449.610a Candicidin vaginal oint-ment.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Candicidin vaginal oint-ment is composed of candicidin and asuitable ointment base. It contains 0.6milligram of candicidin per gram. Its


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21 CFR Ch. I (4–1–98 Edition)§ 449.610b

potency is satisfactory if it is not lessthan 90 percent and not more than 140percent of the number of milligrams ofcandicidin that it is represented tocontain. Its moisture content is notmore than 0.1 percent. The candicidinused conforms to the requirements of§ 449.10(a)(1).



(i) Results of tests and assays on:(a) The candicidin used in making

the batch for potency, loss on drying,pH, and identity.


(ii) Samples required:(a) The candicidin used in making

the batch: 10 packages, each containingapproximately 300 milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.106 ofthis chapter, preparing the sample forassay as follows: Place an accuratelyweighed representative portion of thesample into a separatory funnel con-taining approximately 50 milliliters ofn- hexane (containing 0.1 percentbutylated hydroxyanisole). Shake thesample and n- hexane until homo-geneous. Add 15 milliliters ofdimethylsulfoxide (containing 0.1 per-cent butylated hydroxyanisole) andshake well. Allow the layers to sepa-rate. Remove the bottom layer and re-peat the extraction procedure with asecond 15-milliliter portion ofdimethylsulfoxide (containing 0.1 per-cent butylated hydroxyanisole). Com-bine the extractives in a suitable volu-metric flask and fill to volume withsterile distilled water. Further dilutean aliquot with sterile distilled waterto the reference concentration of 0.06microgram of candicidin per milliliter(estimated).


[39 FR 19134, May 30, 1974, as amended at 40FR 15089, Apr. 4, 1975]

§ 449.610b Candicidin vaginal tablets.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Candicidin vaginal tabletsare tablets composed of candicidinwith suitable binders, diluents, and lu-bricants. Each tablet contains 3 milli-grams of candicidin. Its potency is sat-isfactory if it is not less than 90 per-cent and not more than 150 percent ofthe number of milligrams of candicidinthat it is represented to contain, ex-cept that for the issuance of a certifi-cate for each batch, the candicidin con-tent must be not less than 115 percentand not more than 150 percent of thenumber of milligrams of candicidinthat it is represented to contain. Thetablets shall disintegrate within 30minutes. The loss on drying is notmore than 1 percent. The candicidinused in making the batch conforms tothe standards of § 449.10(a)(1).

(2) Labeling. The drug shall be labeledin accordance with the requirements of§ 432.5 of this chapter.




(b) The batch for potency, loss ondrying, and disintegration time.

(ii) Samples required. (a) Thecandicidin used in making the batch: 10packages, each consisting of approxi-mately 300 milligrams.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.106 ofthis chapter, preparing the sample forassay as follows: Weigh a pool of fivetablets and grind in a mortar to a veryfine powder. Suspend an accuratelyweighed aliquot (of approximately 2grams) in 10 milliliters ofdimethylsulfoxide. Centrifuge for 5minutes at 2,000 revolutions perminute. Carefully decant the super-natant solution into a sterile 250-milli-liter volumetric flask. Wash the resi-due three times with 5-milliliter por-tions of dimethylsulfoxide,centrifuging each time. Add the washesto the 250-milliliter volumetric flask


889


and fill to volume with sterile distilledwater. Using sterile distilled water,further dilute to the reference con-centration of 0.06 microgram ofcandicidin per milliliter (estimated).

(2) Disintegration time. Proceed as di-rected in § 436.212 of this chapter, usingthe method described in paragraph(e)(1) of that section, except use dis-tilled water as the immersion fluid.


§ 449.610c Candicidin vaginal capsules.(a) Requirements for certification—(1)

Standards of identity, strength, quality,and purity. Candicidin vaginal capsulesare gelatin capsules containing 3 milli-grams of candicidin in a suitable andharmless ointment. The candicidincontent is satisfactory if it is not lessthan 90 percent and not more than 150percent of the number of milligrams ofcandicidin that it is represented tocontain. The moisture content is notmore than 0.1 percent. The candicidinused conforms to the requirements of§ 449.10(a)(1).






(ii) Samples required:(a) The candicidin used in making

the batch: 10 packages, each containingapproximately 300 milligrams.

(b) The batch: A minimum of 20 cap-sules.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.106 ofthis chapter, preparing the sample forassay as follows: Remove the tips fromtwo capsules and express the ointmentfrom each capsule into a separatoryfunnel containing approximately 50milliliters of n- hexane (containing 0.1percent butylated hydroxyanisole).Wash out the capsules at least twotimes with 2- to 3-milliliter portions ofwarm (approximately 50° C) n- hexane

(containing 0.1 percent butylated hy-droxyanisole). Add the washes to theseparatory funnel. Shake the sampleand n- hexane until homogeneous. Add15 milliliters of dimethylsulfoxide (con-taining 0.1 percent butylated hydroxy-anisole) and shake well. Allow the lay-ers to separate. Remove the bottomlayer and repeat the extraction proce-dure with a second 15-milliliter portionof dimethylsulfoxide (containing 0.1percent butylated hydroxyanisole).Combine the extractives in a suitablevolumetric flask and fill to volumewith sterile distilled water. Further di-lute an aliquot with sterile distilledwater to the reference concentration of0.06 microgram of candicidin per milli-liter (estimated).


[39 FR 19134, May 30, 1974, as amended at 40FR 15089, Apr. 4, 1975]

§ 449.650 Nystatin vaginal dosageforms.

§ 449.650a Nystatin vaginal tablets.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Nystatin vaginal tablets aretablets composed of nystatin and suit-able and harmless diluents, binders,and lubricants. Each tablet contains100,000 units of nystatin. Its potency issatisfactory if it is not less than 90 per-cent and not more than 140 percent ofthe number of units of nystatin that itis represented to contain. The loss ondrying is not more than 5 percent. Thedisintegration time is not more than 1hour. The nystatin used conforms tothe standards prescribed therefor by§ 449.50(a)(1) (i), (iii), (iv), and (v).





(b) The batch for nystatin content,loss on drying, and disintegration time.

(ii) Samples required:


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21 CFR Ch. I (4–1–98 Edition)§ 449.650b

(a) The nystatin used in making thebatch: 10 immediate containers of ap-proximately 300 milligrams each.


(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Blend a representa-tive number of tablets for 3 to 5 min-utes in a high-speed glass blender withsufficient dimethylformamide to give aconvenient concentration. Dilute an al-iquot with sufficientdimethylformamide to give a stock so-lution containing 400 units of nystatinper milliliter (estimated). Further di-lute the stock solution with 10 percentpotassium phosphate buffer, pH 6.0 (so-lution 6), to the reference concentra-tion of 20 units of nystatin per milli-liter (estimated).


(3) Disintegration time. Proceed as di-rected in § 436.212 of this chapter, usingthe procedure described in paragraph(e)(1) of that section, except use dis-tilled water in lieu of gastric fluid.

[39 FR 19134, May 30, 1974. Redesignated at 43FR 43458, Sept. 26, 1978]

§ 449.650b Nystatin vaginal supposi-tories.

(a) Requirements for certification—(1)Standards of identity, strength, quality,and purity. Nystatin vaginal supposi-tories contain in each suppository100,000 units of nystatin in a suitableand harmless water soluble base. Itspotency is satisfactory if it is not lessthan 90 percent and not more than 130percent of the number of units of nys-tatin that it is represented to contain.Its moisture content is not more than1.5 percent. The nystatin used con-forms to the standards prescribed by§ 449.50(a)(1).








(b) The batch: A minimum of 30 sup-positories.

(b) Tests and methods of assay—(1) Po-tency. Proceed as directed in § 436.105 ofthis chapter, preparing the sample forassay as follows: Place a representativenumber of suppositories into a high-speed glass blender jar containing suf-ficient dimethylformamide to give aconvenient concentration. Blend for 3to 5 minutes. Dilute an aliquot withsufficient dimethylformamide to ob-tain a concentration of 400 units ofnystatin per milliliter (estimated).Further dilute an aliquot with 10 per-cent potassium phosphate buffer, pH 6.0(solution 6), to the reference concentra-tion of 20 units of nystatin per milli-liter (estimated).


[43 FR 43458, Sept. 26, 1978, as amended at 50FR 19920, May 13, 1985]

PART 450—ANTITUMOR ANTIBIOTICDRUGS


Sec.450.10a Sterile bleomycin sulfate.450.20 Dactinomycin.450.22 Daunorubicin hydrochloride.450.24 Doxorubicin hydrochloride.450.30 Idarubicin hydrochloride.450.40 Plicamycin.450.45 Mitomycin.

Subpart B [Reserved]

Subpart C—Injectable Dosage Forms

450.210 Sterile bleomycin sulfate.450.220 Dactinomycin for injection.450.222 Daunorubicin hydrochloride for in-

jection.450.224 Doxorubicin hydrochloride

injectable dosage forms.450.224a Doxorubicin hydrochloride for in-

jection.450.224b Doxorubicin hydrochloride injec-

tion.450.230 Idarubicin hydrochloride for injec-

tion.450.240 Plicamycin for injection.450.245 Mitomycin for injection.


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ANTIBIOTIC DRUGS Subpart Aâ€”Bulk Drugs - GPO

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