ANALYSIS OF SULFONAMIDES

ANALYSIS OF SULPHONAMIDES 12/15/20161VIKAS COLLEGE OF PHARMACY BY @@@ Mr.S @@@ Pharmaceutical Analysis & Quality Assurance, Vikas College Of Pharmacy, Vissannapeta.

CONTENTS12/15/20162INTRODUCTIONCLASSIFICATIONMECHANISM OF ACTIONMETHOD OF ANALYSISCONCLUSIONREFERENCESVIKAS COLLEGE OF PHARMACY

IntroductionSulphonamide is derived from Prontosil,a prodrug that is metabolised invivo by Azoreductase.Anti bacterial drugs used in the treatment of infections caused by bacteria (gram positive and gram negative bacteria).Available in the form of tablets, suspensions,parenterals, opthalmic solutions,ointments.12/15/20163

VIKAS COLLEGE OF PHARMACY

Classification12/15/20164SulphadiazineSulphathiazolesulphamethoxazolesulphadoxineVIKAS COLLEGE OF PHARMACY

Structures of Sulphonamides12/15/20165

VIKAS COLLEGE OF PHARMACY

Mechanism of action

12/15/20166Sulfonamides (such as sulfamethoxazole) and diaminopyrimidines (such as trimethoprim) inhibit different enzymes in the biosynthesis of tetrahydrofolate in the bacteria. Due to the inhibited production of tetrahydrofolate, the bacteria is unable to synthesize the thymidine, and is therefore also unable to produce new DNA or RNA. This eventually leads to the death of the bacteria.VIKAS COLLEGE OF PHARMACY

6

Mechanism of action12/15/20167

VIKAS COLLEGE OF PHARMACY

Analytical methods for sulfadrugs

Titrimetric method : a)Diazotization titration b)Nonaqueous titration c)Argentometric titrationUV-spectrophotometryColorimetryChromatographyFluorimetry

12/15/20168VIKAS COLLEGE OF PHARMACY

8

SULPHADIAZINE12/15/20169Chemical formula:C12H14N4O2SIUPAC name:4-amino-N-(4,6-dimethylpyrimidin-2-yl)benzene-1-sulfonamideIndication :For the treatment bacterial infections causing bronchitis, prostatitis and urinary tract infections.Do not take calcium, aluminium, magnesium or iron supplements within 2 hours of taking this medication.

VIKAS COLLEGE OF PHARMACY

DiazotizationWhen aromatic primary amines with nuclear NH2 groups can be determined quantitatively by standard sodium nitrite solution required to convert them into diazonium salts.12/15/201610

VIKAS COLLEGE OF PHARMACY

SULPHATHIAZOLE12/15/201611

Chemical Formula C9H9N3O2S2IUPAC Name 4-amino-N-(1,3-thiazol-2-yl)benzene-1-sulfonamideIndication: effective against a wide range of gram positive and gram negative pathogenic microorganisms. Although no longer used in humans, it is used in cattle.VIKAS COLLEGE OF PHARMACY

Non-Aqueous TitrationsFor determination of weak acids and weak basesTYPESAcidimetry in non aqueous titration Alkalimetry in non aqueous titrationThe sulfonamides contain weak acid group, so we are using alkalimetry in NAT.In these titration:Titrants used :Sodium methoxide,potassium methoxide,Lithium methoxide.

12/15/201612VIKAS COLLEGE OF PHARMACY

Solvents: a. Strong base solvents: n-butyl amine ,Morpholine b. Weak base solvents: DMF,Anhydrous pyridineIndicators: 1. 0.1N Alkali Methoxide Indicator 2. Thymol Blue IndicatorREAGENTS:0.1N alkali methoxide.Ex:CH3ONa:

12/15/201613VIKAS COLLEGE OF PHARMACY

ASSAY PROCEDURE

12/15/2016142. Thymol blue indicatorVIKAS COLLEGE OF PHARMACY

14

Argentometric TitrationPRINCIPLE:Argentometric titration is otherwise known as Precipitation titrationArgentometry involves the use of the standard solution of silver nitrate as the titrant for estimation of the halides(chlorides,bromides & iodides)12/15/201615

VIKAS COLLEGE OF PHARMACY

PROCEDURE

12/15/201616Whole mixture is allowed to stand in darkThe ppt is collected on double fold filter paperWash ppt with waterFilterate is acidifed with HNO3Excess AgNO3 is titrated with 0.1N Ammonium thiocyanate

Using ferric alum indicatorVIKAS COLLEGE OF PHARMACY

procedure:Take 0.2-0.3g of sulfonamidesDissolve in 0.1 N NaOHSOLUTION IS ADJUSTED TO GIVE BLUE COLOUR WITH THYMOPHTHALINEVolume make upto 50ml with distilled waterAdd 0.1 n H2SO4 to discharge blue colourAdd 25ml of 0.1N AgNO3Whole mixture is allowed to stand in darkThe ppt is collected on double fold filter paperWash ppt with waterFilterate is acidify with HNO3Excess AgNO3 is titrated with 0.1N AMMONIUM THIOCYNATE USING Ferric alum indicator16

SULPHAMETHAZINE12/15/201617

Chemical Formula: C12H14N4O2SIUPAC Name: 4-amino-N-(4,6-dimethylpyrimidin-2-yl)benzene-1-sulfonamideIndication:For the treatment bacterial infections causing bronchitis, prostatitis and urinary tract infections.

VIKAS COLLEGE OF PHARMACY

UV-SpectrophotometryEstimation of sulfamethazine:

12/15/201618

VIKAS COLLEGE OF PHARMACY

Take sulfamethizine sample dissolved in dil.H2SO4Add small amount of resorcinolAbsofbance measure at 320 nm.18

12/15/201619Unknown should be compared with standard of same concentration as the system obeys Beers law.Absorbance measured at 545nm.Sulfonamide(mg)=Absorbane of sample standard(mg) absorbance of standard

VIKAS COLLEGE OF PHARMACY

Colorimetry12/15/201620Analysis of sulfa drugs is done by converting primary amino group of sulfonamides into a diazonium salt by diazotisation and later coupling with suitable chromogenic agent to form Azo dye.

Automated ColorimetryVIKAS COLLEGE OF PHARMACY

12/15/201621Chromogenic Reagents

Para dimethyl amino benzaldehyde methodDiazotisation followed by coupling with BM Reagent-1,2-naphthaquinone-4-sulphonate sodium(folins reagent)Thiobarbituric acid method-napthol method VIKAS COLLEGE OF PHARMACY

Para dimethyl amino benzaldehyde methodBrotton-marshall reagent1,2-naphthaquinone-4-sulphonate sodiumThiobarbituric acid method-napthol methodDiazotisation followed by addition of alkali21

PDAB (Para Dimethyl Amino Benzaldehyde) methodDrug solution is treated with solution of PDAB under acidic conditions results in formation of AZOMETHINE,a yellow colour chromogen.Measured at max :440nm.It is an example of schiffs base formation22

R1=HR2=C6H5-N(CH3)2R3=C6H5SO2NH-RYELLOW SCHIFFS BASE PDABSULPHONAMIDE12/15/2016VIKAS COLLEGE OF PHARMACY

1)Drug solution is treated with solution of PDAB under acidic conditions results in formation of yellow colour chromogen with max 440nm.2)It is an example of schiffs base formation

22

Diazotisation Followed By Coupling With BM Reagent

NH2SO2

DIAZONIUM SALT

N2cl

NHCHCHNH2

NHCH2CH2NH2SO2NH2BM REAGENT(N-1-NAPHTHYL-ETHYLENE DIAMINE DIHYDRO CHLORIDE) N=N12/15/201623PINK AZODYEVIKAS COLLEGE OF PHARMACY

N2+cl-SO2NH2

N=NNHCH2CH2NH2BM REAGENT23

CHROMATOGRAPHIC METHOD12/15/201624Paper chromatography is most widely used for analysis of sulfa drugs.The mobile phase used is a mixture of equal volumes (1:1) of 3%NH4OH and n-butanol.PAPER CHROMATOGRAPHY

Paper ChromatographyVIKAS COLLEGE OF PHARMACY

12/15/201625Thin Layer Chromatography is also most widely used for analysis of sulfa drugs in agricultural industries.Mobile phase is chloroform and tert-butanol in ratio of 80:20THIN LAYER CHROMATOGRAPHY

TLC platesVIKAS COLLEGE OF PHARMACY

FLUORIMETRY 12/15/201626

FluorimeterVIKAS COLLEGE OF PHARMACY

.Then it is coupled with 2,4,6-Triaminopyridine.The product obtained in the above reaction is subjected to oxidation to produce triazo derivative which has flouresence property and can be analysed by spectro flourimetre.26

CONCLUSION I here by conclude that there is a much necessity for the study of analytical methods of Sulphonamides as they are widely used class of drugs as Anti bacterial agents..12/15/201627VIKAS COLLEGE OF PHARMACY

REFERENCESThe Indian Pharmacopoeia1996P. D. Sethi, Quantitative Analysis of Drugs in Pharmaceutical Formulations,. B. Morelli, J. Pharm. Biomed. Anal.,1989, 7, 577.P. B. Issopoulos, Acta. Pharm. Hung.,1992, 6, 3138.M. Knochen, J. Giglio and B.F. Reis, J. Pharm. Biomed. Anal., 2003, 33, 191. K.A Connors :Text book of Pharmaceutical Analysis,Third edition,Page no:(62-63Higuchi,Beckmman&Hassan :Pharmaceutical Analysis,second editiom,Page no:(137-157)V.N Rajasekharan,Text book of Pharmaceutical In Organic Chemistry,Page no(126-128)12/15/201628VIKAS COLLEGE OF PHARMACY

12/15/201629

VIKAS COLLEGE OF PHARMACY