S.

NO

NAME

STRUCTURE

QUALIFICATION

QUANTIFICATION

(1).Diaminopyrimidines 1. Trimethoprim (IP,BP,USP) A. Infrared spectroscopy. B. UV absorbance at 287nm. C. 25mg+ 2ml of 1.6%w/v of KMNO4 heat add 0.4ml of HCHO + 1ml of 0.5M H2SO4. Cool and filter.Add 2ml of CHCl3 .Chloroform layer shows green fluorescence under UV at 365nm. 2. Pyrimethamine (IP,BP,USP) A. Infrared spectroscopy. B.Thin Layer Chromatography ( for related substances) C. UV Spectroscopy Potentiometric titration(0.1MHClO4)

Potentiometric. titration.(0.1M HClO4)

(max. wavelength at 272nm.

(2)Quinolones (i). Ciprofloxacin (IP,BP,USP) A. Infrared Spectroscopy B. Thin Layer Chromatography Liquid chromatography. (mobile phase:0.025M Phosphoric Acid and ACN 87:13) Flow rate :1.5ml/min Detection wavelength:278nm (ii) Nalidixic acid (IP,BP,USP) A. Infrared Spectroscopy. B.Absorbance ratio method. (A 0.0005 %w/v solution in 0.1M sodium hydroxide shows absorption maxima at about 258 nm and Potentiometric titration (0.1M Ethanolic NaOH)

334 nm) C. Thin Layer Chromatography(for related substances) D. Addition of 0.5ml of a 10%w/v of 2naphthol in ethanol to 0.1g of drug gives orange red colour. A. Infrared spectroscopy B. UV Absorbance at 273 nm in 0.0005%w/v in 0.1N NaOH.

(iii)

Norfloxacin (IP,BP)

Potentiometric titration(0.1MHClO4) Non aqueous titration.

(3)Tetracyclines (i) Oxytetraycline dihydrate (IP,BP,USP) A. Thin Layer chromatography B. Addition of H2SO4 gives colour change from red to yellow. C. 2mg of drug+2ml of diazotized

Microbiological Assay (Method A or B)

sulphanilic acid gives orange red to brownish red colour.

(ii)

Doxycycline Hydrochloride (IP,USP)

A. Infra red Spectroscopy B. Thin Layer Chromatography C. Addition of H2SO4 gives yellow colour D. A 5%w/v of solution gives the reaction of chlorides

Liquid chromatography Mobile phase: 60gms of 2methyl-2-propanal in 200ml of water + 400ml of phosphate buffer(PH 8) + 50ml of 1%w/v tetrabutyl ammonium dihydrogen phosphate (PH 8) + 10ml of 4%w/v disodium edentate(PH 8) make up to 1 litre.

REACTION OF CHLORIDES: A. Dissolve drug equivalent to 2mg of Chloride ion in 2ml in water. Acidify with dil.HNO3 add 0.5ml AgNO3.A curdy white ppt is formed which is insoluble in nitric acid, soluble in dil.NH3.Addition of dil.HNO3 results in reprecipitation. B. Dissolve drug equivalent to 10mg of chloride ion add 0.2g of K2Cr2O7 and 1ml of H2SO4 .Place a filter paper strip moistened with 0.1ml of diphenyl carbazide solution paper turns violet red colour.

(4)Nitro benzene derivatives (i) Chloramphenicol (IP,BP,USP) A. Infra red spectroscopy B.Thin Layer Chromatography (For related substances) C.10mg drug+50mg of zinc+0.1gm of anhydrous CH3COONa+0.1ml benzoylchloride+0.5 ml FeCl3 6.H2O gives red violet to purple colour. If zinc is omitted no colour. D.50mg of drug+2ml of ethanolic KOH gives reaction of chlorides.

A1%1c.m. method. Max.wavelength(278nm) A1%,1cm value= 297.

(5)Amino glycosides (i) Streptomycine sulphate (IP,BP,USP) A. Thin Layer Chromatography B.5-10mg of drug+1M NaOH+2M HCl+10%w/v FeCl3 gives violet colour. C.10mg+1M HCl heat add 2ml of napthol a faint yellow colour is observed. D. Gives reaction of Sulphates. (ii) Gentamicin sulphate (IP,BP,USP) A. Thin Layer Chromatography B. 10mg +1ml water+5ml of40%w/v H2SO4 scaned from Microbiological 240-330nm no assay(method A) absorbtion maxima is seen. C. Finger print Analysis Micro biological assay (method A or B)

(Chromatography) D. Gives reaction A of sulphates

REACTION OF SULPHATES: A. Dissolve about 50 mg of the substance under examination in 5 ml of water or use 5 ml

of the prescribed solution. Add 1 ml of dilute hydrochloric acid and 1 ml of barium chloride solution; a white precipitate is formed.
B. Add 0.1 ml of iodine solution to the suspension obtained in test A; the suspension

remains yellow (distinction from sulphites and dithionites) but is decolorised by adding, dropwise, stannous chloride solution (distinction from iodates). Boil the mixture; no coloured precipitate is formed (distinction from selenates and Tungstates.
C. Dissolve about 50 mg of the substance under examination in 5 ml of water or use 5 ml

of the prescribed solution. Add 2 ml of lead acetate solution; a white precipitate, soluble in ammonium acetate solution and in sodium hydroxide solution, is produced.

(6) Macrolide Antibiotics

(i)

Erythromycin (IP,BP,USP)

A. Infrared Spectroscopy. B. Thin Layer Chromatography. C.5mg of drug +0.02%of xanthydrol and heat red colour is produced. D. Addition of 5mlof 7M HCl develops yellow colour. Microbiological assay(method A)

(ii)

Roxithromycin (IP,BP)

A.Infrared Spectroscopy.

Liquid Chromatography (mobile phase 30(ACN):70(4.8%w/vam monium dihydrogen phosphate)

B. Chromatography.

(7)Poly Peptide Antibiotics (i) Bacitracin (IP,BP,USP) A. Thin Layer Chromatography. B. 5mg of drug+1ml of 0.2%w/v of

Microbiological Assay(method A)

ninhydrin in 1butanol +0.5ml of pyridine gives deep purple colour.

(8)Nitro furantoin derivatives (i) Nitrofurantoin (IP,BP) A. UV Absorbance Ratio Method (absorbance maxima at 266 and 367 nm) B. 1ml of 0.1%w/v in dimethyl formamide +0.1mlof 0.5MethanolicKOH gives brown colour (ii) Furazolidone (IP,BP,USP) B.1mg+1ml of dimethyl formamide+0.05 A. Infrared Spectroscopy. A1%,1cm method. (A1%,1cm=750) Wavelength:367 nm

A1%,1cm method ( A1% ,1cm =765) Wavelength: 367nm

ethanolic KOH gives blue colour.

(carry out the procedure protected from sunlight).

(9)Nitro imidazoles (i) Metronidazole (IP,BP) A. Infrared Spectroscopy. B. Absorbance Ratio Method (max.abs :277nm Min.abs : 240nm) C. 10mg of drug+1010mg of zinc+1ml of water+0.25ml of2M HCl gives reaction of primary aromatic amines (ii) Tinidazole (IP,BP) A.Infrared Spectroscopy B.0.001%w/v in Potentiometric titration using 0.1MHClO4

Titration 0.1MHClO4 using nile blue as

methanol shows absorbance max.310nm. C.5mg of drug+ 0.1%HCl+50mg of zinc+4ml of 1%w/v vanillinheat and cool gives greenish yellow colour.

indicator.

REACTION OF PRIMARY AROMATIC AMINES:

Acidify the prescribed solution with 2 M hydrochloric acid or dissolve 0.1 g of the substance under examination in 2 ml of 2M hydrochloric acid and add 0.2 ml of sodium nitrite solution. After 1 or 2 minutes add the solution to 1ml of 2-naphthol solution; an intense orange or red colour and, usually, a precipitate of the same colour is produced.

(10)Nicotinic acid derivatives (i) Isoniazid (IP,BP,USP) A. Infrared Spectroscopy. B.0.1gm in water+0.1gm of vanillin yellow ppt is formed whose MP is 226 0 - 2310 C. Melts at 1701740C (ii) Pyrazinamide (IP,BP,USP) A. Infrared Spectroscopy B.UV Spectroscopy (absorbance ratio method ) (Absorbances at 268nm and 310nm) C. Boil 20mg+5ml of NaOH ,NH3 is Titration with 0.1M NaOH. Indicator : methyl red.

Liquid chromatography

evolved recognized by odour. (iii) Ethionamide (IP,BP,USP) A. Infrared Spectroscopy B.10mg+5ml of 0.1M AgNO3 gives dark brown ppt. C.Melting point 1581640. Liquid chromatography