abhi-atb : IMA : NCP

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WEL – COME To Seminar on AntiTubercular Drugs

ANTI TUBERCULAR DRUGS : 

ANTI TUBERCULAR DRUGS Presented by : Abhijith.P 1st yr M Pharm Pharmaceutical Analysis Under the Guidance of B.Gopinath, Pharmaceutical Analysis

ANTI TUBERCULAR DRUGS : 

ANTI TUBERCULAR DRUGS

RIFAMPIN : 

RIFAMPIN Chemical name : 2,7-(epoxypentadeca [1,11,13] trienimino)naphtho [2,1-b]furan-1,11(2H)-dione 5,6,9,17,19,21-hexahydroxy-23-methoxy-2,4,12,16,18,20,22-heptamethyl-8-[N-(4-methyl-1-perazinyl)formimidoyl]-21-acetate. Chemical Structure : Chemical formula : C43H58N4O12 Mol. Wt. : 822.95

Physical properties : 

Physical properties It is a Red-Orange, Odourless Crystalline Powder. Freely soluble in Chloroform, Soluble in Methanol, Ethanol(95%) and Insoluble in n-butanol, Cyclohexane. It’s Melting point range’s b/w 1830C - 1880C

Analytical Methods : 

Analytical Methods 1. Paper Chromatography 2.Thin Layer Chromatography 3. Column Chromatography 4. HPLC 5. Fluorimetric Method

PAPER CHROMATOGRAPHY : 

PAPER CHROMATOGRAPHY Here…….Descending- Paper-Chromatography is Carried out. Stationary Phase : Whatmann-3mm-Filter Paper Mobile Phase : Methanol : n-Octanol (4:1) (PH : 6) Detection : The intensity of Red-Orange spots was measured by an Analytrol Photodensiotometer (Mod : R13-450nm filter)

THIN LAYER CHROMATOGRAPHY : 

THIN LAYER CHROMATOGRAPHY TLC is mainly developed for the analysis of Rifampin & it’s Metabolites in body fluids. The Colour spot was eluted & measured Spectrophotometrically

COLUMN CHROMATOGRAPHY : 

COLUMN CHROMATOGRAPHY This method is mainly used for Determination of Rifampin & it’s Metabolites in Urine, Bile & Serum by extraction with Chloroform & by Column chromatography followed by Spectrophotometry The Liquid-Solid Chromatography is carried out using a Glass column of 7cm (4mm.i.d) long. The column is packed with Silica Gel-G buffered at PH=6 & Chloroform.

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Chloroform-Methanol is used as Solvents in progressively increasing amounts(5%, 10% & 16.6%) Flow Rate : 0.15ml/min. Rifampin & it’s Metabolites were than quantitated by reading the absorbance of the elutes at 475 nm.

HPLC : 

HPLC Rifampin & 35 desacetylrifampin were separated under the following condition : Dupont 820 Chromatograph equipped with UV-detector Column used : ODS Permaphas column. Temp of Column : 500C at 1000psi Mobile Phase used : Water : Methanol

FLUORIMETRY : 

FLUORIMETRY Rifampin was determined Fluorimetrically by transforming it with Hydrogen peroxide into a Fluorescent product The maximum fluorescence develop in an Aq.Carbonate-bicarbonate buffer at PH=9.2 at 480nm The excitation wave length is 370nm. The Fluorescence intensity is linear in the conc. range from 0.1-10mcg.

ETHAMBUTOL HYDROCHLORIDE : 

ETHAMBUTOL HYDROCHLORIDE Chemical name : N, N1-di-isopropyl ethylenediamine hydrochloride. Chemical formula : C10H24O2N2 (2HCl) Chemical structure : . 2HCl Mol. Wt. : 277.5g/mol

Physical properties : 

Physical properties It is a White, Odourless, Bitter Tasting, Crystalline Powder. Easily soluble in Water, Sparingly soluble in Ethanol. It’s Melting point range’s b/w 198.50C – 202.50C

Analytical Methods : 

Analytical Methods 1. Colorimetry 2. Non-Aqueous Titration

COLORIMETRY : 

COLORIMETRY Take the sample solution Add bromothymol blue Complex will form Give yellow color Determined at 520 nm

NON AQUEOUS TITRATION : 

NON AQUEOUS TITRATION Take the sample dissolve CH3COOH add Mercuric acetate add Crystal violate Titrate with 0.1M HClO4

CYCLOSERINE : 

CYCLOSERINE Chemical name : 4-Amino-3-Isoxazolidone Chemical formula : C3H6N2O2 Chemical structure : Mol. Wt. : 102.09g/mol.

Physical properties : 

Physical properties It is a White to slightly Yellow , Odourless Crystalline powder. It is Insoluble in common organic solvents, but readily Soluble in Water. It’s Melting point range’s b/w 1540C - 1560C

Analytical Methods : 

Analytical Methods Spectrophotometry. Colorimetry Paper Chromatography. Thin Layer Chromatography

SPECTROPHOTOMETRIC METHOD : 

SPECTROPHOTOMETRIC METHOD Cycloserine has been determined by a Spectrophotometric method based on the complex formation with chloranil. The method involves addition of cycloserine solution to chloranil reagent in borate buffer of pH 9; and heating at 65°C for 45 min. The complex formed exhibits absorption maximum at 348 nm

COLORIMETRY : 

COLORIMETRY This method is mainly used for determination of Cycloserine in Biological fluids such as Blood, Urine & CSF . Cycloserine reacts with Sodium-nitropentacyano- ferrate in a slightly acidic aqueous soln to give an intense Blue colour complex which is quantitatively measured at 625nm

PAPER CHROMATOGRAPHY : 

PAPER CHROMATOGRAPHY

THIN LAYER CHROMATOGRAPHY : 

THIN LAYER CHROMATOGRAPHY Detecting Agent : Ninhydrin spray reagent

REFERENCES : 

REFERENCES Text Book of Pharmacology by Triphati Analytical Profiles of Drug Substances by Klaus Florey Instrumental Analysis by Skoog Text Book of Pharmaceutical Analysis by Higuchi Internet Source

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