Assay-A

S.#	Assay Method (A)
i	Assay Method of  Aceclofenac Tablet
	Solvent Methanol Standard 50 mg in 100 ml take 2 ml in 50 ml (Shake 30 minutes) Sample Same as standard. Take absorbance at 276 nm.
ii	Assay Method of  Albendazole Tablet
	Sample                                                                                                                                                                                                                         Dissolve powdered sample equivalent to about 100 mg of Albendazole in a 60 ml previously neutralized glacial acetic acid using crystal violet solution as indicator. Titrate with 0.1M perchloric acid and determine the end point.Each ml of 0.1M perchloric acid VS is equivalent to 26.53 mg of Albendazole. % of content = V x F x 26.53 x 100 / weight of Albendazole in mg
iii	Assay Method of Amlodipine Besylate + Valsartan Tablet
	Buffer 1.74 gm of K2HPO4 was dissolved in1L of deionized water and pH was adjusted to 3.6 by ortho-Phosphoric acid. Ratio   Degassed mixture of Phosphate Buffer (pH 3.6, 0.01M) : ACN : Methanol = 50% : 40% : 10% Diluents: Methanol Standard Transfer an accurately weighted quantity of about 10 mg of Valsartan WS, 7 mg of Amlodipine Besilate WS (equivalent to 5.049 of Amlodipine) into a 100 ml volumetric flask then add 70 ml of solvent, shake for 10 minutes and sonicate for 30 minutes. Dilute with solvent to volume and mix well .Filter this solution with 0.45 m membrane filter before injection. (Concentration of Valsartan 0.08 mg/ml and Concentration of Amlodipine 0.05049 mg/ml). 1 mg Amlodipine ≡ 1.3864 mg of Amlodipine Besilate. Sample Weigh and transfer about 275 mg powder equivalent to 160 mg of Valsartan & 5 mg of Amlodipine into a 100ml volumetric flask then add 70 ml of solvent, shake for 10 minutes and sonicate for 30 minutes. Dilute with solvent to volume and mix well .Filter this solution with whatman filter paper. Filter this solution with 0.45 m membrane filter before injection. (Concentration of Valsartan 3.2 mg/ml and Concentration of Amlodipine 0.05 mg/ml). Condition Waters Spherisorb 5mm ODS2   4.6 ´ 250 mm./ 40oC / UV at 240 nm / 1 ml per minute / 10 mL / 15 minutes. 7.5 minutes for Valsartan & 10.5 minutes for Amlodipine Besilate approximately.
iv	Assay Method of Amoxicillin Trihydrate Tablet, Capsule & PFS
	Buffer Dissolve 173 g of Sodium Hydroxide & 20.6 g of Sodium Acetate in sufficient water to produce 1000 ml. Hydroxylamine Hydrochloride solution Dissolve 350 g of hydroxylamine hydrochloride in sufficient water to produce 1000 ml. Neutral Hydroxylamine solution Mix 1 volume each of Hydroxylamine solution and the Buffer solution. Check the pH & if necessary adjust the pH to 7.0   0.1 by adding an additional amount of one of the component. To 1 volume of this neutral solution add 8 volume of water & 2 volume of 95% ethanol. This solution must be freshly prepared. Ferric Ammonium Sulfate Solution Dissolve 272 g Ferric Ammonium Sulfate in a mixture of 26 ml of concentrate Sulfuric Acid and sufficient water to make 1000 ml. This solution may be used for 1 week when stored in a brown bottle in room temperature. Standard Dissolve Amoxicillin Trihydrate WS equivalent to 31.25 mg of Amoxicillin in 2% w/v Sodium Bi-Carbonate solution and dilute to 25 ml with the same solvent. Sample Take powder equivalent to 125 mg of Amoxicillin in a 100 ml volumetric flask, add 70 ml of 2% w/v Sodium Bi-Carbonate solution and shake for about 20 minutes, then volume to 100 ml with 2% w/v Sodium Bi-Carbonate solution  mix and filter. Procedure Take 2 ml solution from each standard and sample in two separate 25 ml volumetric flask. Add 2 ml of water and 2.5 ml of Neutral Hydroxylamine Solution, mix & allow to stand for 5 minutes. Add 2.5 ml Ferric ammonium sulfate solution and mix.                                For Blank mix 2 ml of 2% w/v Sodium Bi-Carbonate solution with 2 ml water, 2.5 ml Neutral Hydroxylamine Solution and 2.5 ml Ferric Ammonium sulfate solution.               After 3 minutes determine the absorbance of the resulting solutions at 480 nm.
v	Assay Method of Ascorbic Acid Tablet & Syrup
	Sample Accurately Transfer the 5.0 ml Syrup equivalent to 100.0 mg of Ascorbic Acid using by 5ml glass pipette which is previously ringed by sample syrup in a mixture of 20 ml of distilled water & 15 ml of 1.0 M Sulphuric Acid to the Conical flask & shake at least for 5 minutes to dissolve. Then add to 0.5ml Ferroin Solution as indicator and mix well. Carry out a potentiometric titration using 0.1 M Ammonium Cerium (IV) Sulphate VS. Each ml of 0.1 M Ammonium Cerium (IV) Sulphate VS is equivalent to 8.806 mg of Ascorbic Acid.
vi	Assay Method of Aspirin Tablet
	Sample To a quantity of the powder containing 0.5 g of Aspirin add 30 ml of 0.5M sodium hydroxide VS, boil gently for 10 minutes and titrate the excess of alkali with 0.5M hydrochloric acid VS using phenol red solution as indicator. Repeat the operation without the substance being examined. The difference between the titrations represents the amount of sodium hydroxide required (V). Each ml of 0.5M sodium hydroxide VS is equivalent to 45.04 mg of Aspirin. Calculation: % of content =   V x F x 45.04 x 100 / W
vii	Assay Method of Atenolol Tablet
	Sample Transfer the powder equivalent to 50 mg of Atenolol in a 100-ml flask, add 60 ml of methanol, heat the resulting suspension to 60° and shake for 15 minutes. Cool, dilute to 100 ml with methanol, filter and Dilute 10 ml of the filtrate to 50 ml with methanol. Measure the absorbance of the resulting solution at the maximum at 275 nm using methanol as blank 53.7 % of content =     Absorbance of Sample x 100 / Concentration of Sample x 53.7
viii	Assay Method of Atorvastatin Tablet
	Buffer Dissolved about 1.36 g of Potassium Dihydrogen Ortho Phosphate (KH2PO4) in 900 ml purified water, mix well. Adjust pH to 3.2 ± 0.5 with phosphoric acid and make volume with water up to 1000 ml. Ratio   Buffer pH 3.2: Acetonitrile = 45: 55 Solvent: Methanol Standard 25 mg*50 ml => take 4 ml*25 ml  (Shake 30 minutes) Sample Same as standard. Condition Waters XTerra RP 18  5 mm 4.6 ´ 250mm./ 40º C / UV at 235 nm / 10 minutes
ix	Assay Method of Azithromycin Tablet
	M. Phase 34.84 g/L solution of Dipotassium Hydrogen Phosphate (previously adjusted to pH 6.5 with Phosphoric acid): Acetonitrile: Water = 10: 35: 55 Diluents   Acetonitrile: Water = 60: 40                                                   Standard 100 mg*25 ml (Shake 20 minutes) Sample Same as standard. Condition Waters XTerra RP 18 5 mm 4.6 ´ 250mm. / 70º C / UV at 215 nm / 30 minutes
x	Assay Method of Aceclofenac Tablet
	Solvent            Methanol Standard        50 mg*100 ml*take 2 ml*50 ml  (Shake 30 minutes) Sample            Same as standard. Take absorbance at 276 nm
xi	Assay Method of Allopurinol Tablet
	Sample            Weigh and powder 20 tablets. Shake a quantity of the powder containing 100 mg of Allopurinol with 20 ml of 0.05M sodium hydroxide for 20 minutes, add 80 ml of 0.1M hydrochloric acid, shake for 20 minutes, and add sufficient 0.1M hydrochloric acid to produce 250 ml. Sonicate for 10 minutes. Filter the solution and dilute 2 ml of the filtrate to 50 ml with 0.1M hydrochloric acid. Measure the absorbance of the resulting solution at the maximum at 250 nm, using 0.1M hydrochloric acid in the reference cell. Calculation % of Allopurinol = Absorbance of Sample ´ 100 / Conc. Of spl. ´563
xii	Assay Method of Almitrine Bismesylate Tablet
	Mobile Phase: Methanol 85%: Water 15% Diluents:           Methanol Standard:         25 mg*100 ml (Shake 15 minutes) Sample:             25 mg*100 ml (Shake 15 minutes) Condition       ODS2 5 µ 4.0 mm x 250 mm /220 nm /0.8 ml per minute/10 micro liter / 40°C /10 minutes (6 minutes approx.)
xiii	AssayMethod of Alprazolam Tablet
	Mobile Phase Methanol 90%: Distilled Water 10% Diluents          Mobile Phase Standard        25 mg*100 ml* take 5 ml* 25 ml (Shake 20 minutes) Sample            Same as standard. Condition       ODS2 5 µ 4.6 mm x 250 mm Analytical Column. / UV at 254 nm / 1 ml per minute / 20 micro liter / 40°C /10 minutes.
xiv	Assay Method of Aluminium Oxide Tablet
	Test preparation        Take powder equivalent to 200 mg of aluminum oxide in a 100 ml volumetric flask. Add 7ml of Concentrated Hydrochloric acid & heat gently till gives a clear solution. Cool. Add water to make volume 100 ml. Filter, if necessary. Sample            Take 10 ml of the filtrate to a 250 ml conical flask. Add in the order named & continuous stirring 20 ml of 0.05 M Na2EDTA VS & 20 ml of acetic acid – ammonium acetate buffer TS. Then heat the solution near the boiling point for 5 minutes. Cool and add 50 ml of alcohol and 2 ml of dithizone TS. Titrate the solution with 0.05 M ZnSO4 VS until a bright rose pink color produced. Each ml of 0.05 M Na2EDTA VS º 2.549 mg of Al2O3 % of Aluminium Oxide = (20 ´ F1 - Volume of 0.05 M ZnSO4 ´ F2) ´2.549 ´ 100 / Amount of Al2O3 taken in 10ml solution (mg)                Where F1 & F2 are the factor of 0.05M Na2EDTA VS & 0.05M ZnSO4 VS respectively.
xv	Assay Method of Ambroxol Hydrochloride Solution
	Buffer Dissolve 1.32 gm of Di -ammonium Hydrogen Phosphate into 1000 ml volumetric flask & add 950 ml distilled water, adjust the pH 7.0 ± 0.05 with 9 % Phosphoric Acid TS & make volume with distilled water up to 1000 ml Ratio               Acetonitrile: Buffer = 60: 40           Solvent            Methanol                                                                   Standard        15 mg ----® 100 ml ----® take 5 ml ----® 25 ml (Shake 30 minutes) Sample            Same as standard Condition       ODS2, 10 m 4.6 x 250 mm / 400 C/ UV at 248 nm/15 minutes
xvi	Assay Method of Aminophylline Injection
	Sample            To a volume containing 100 mg of Aminophylline add sufficient 0.01 M Sodium Hydroxide to produce 250 ml. Dilute 5 ml to 250 ml with 0.01 M sodium Hydroxide and measure the absorbance of the resulting solution at the maximum at 275 nm. Calculate the content of Theophylline taking 650 as the value of A (1%, 1cm) at the maximum at 275 nm. Calculation     % of Aminophylline = Absorbance of Sample´100/Conc.Of spl.´ 650
xvii	Assay Method of Amitriptyline Hcl Tablet
	Solvent            0.1 Hcl Standard        50 mg*100 ml => take 2 ml*100 ml (Shake 20 minutes) Sample            Same as standard. Measure the absorbance of both sample & standard solution at 239 nm using 0.1 M Hcl as blank.
xviii	Assay Method of Amlodipine Besylate Tablet
	Solvent            Methanol Standard        25 mg*00 ml => take 2 ml*50 ml (Shake 20 minutes) Sample            Same as standard. Measure the absorbance of both sample & standard solution at 237 nm using methanol as blank.
xix	Assay Method of Amlodipine Besylate + Atenolol Tablet
	Buffer Transfer accurately weighed about 2.72 gm of Potassium Dihydrogen phosphate with 950 ml distilled water into 1000 ml volumetric flask & add 5 ml of Triethylamine & mix well. Adjust the pH 3.0 ± 0.05 with phosphoric acid & make volume with distilled water up to 1000 ml. Ratio                            Acetonitrile: Buffer º 40: 60 Diluents          Acetonitrile: Buffer º 40: 60 Standard        50 mg Atenolol & 25 mg Amlodipine besylate*50 ml => take 5 ml*50 ml (Shake 20 minutes) Sample            Same as standard. Condition       Waters Spherisorb 5mm ODS2   4.6 ´ 250mm. / UV at 274 nm & 237 nm / 11 minutes. Atenolol: 2.5 minutes and Amlodipine Besylate: 7.8 minutes (approximately).
xx	Assay Method of Amlodipine Besylate + Atorvastatin Calcium Tablet
	Buffer Transfer accurately weighed about 2.72 gm of Potassium Dihydrogen phosphate with 950 ml distilled water into 1000 ml volumetric flask & add 5 ml of Triethylamine & mix well. Adjust the pH 3.0 ± 0.05 with phosphoric acid & make volume with distilled water up to 1000 ml. Ratio               Mobile phase (A)   : Buffer pH 2.5  Mobile Phase (B)   : Acetonitrile Diluents                 : Water (30%):  Acetonitrile (70%) Standard        Accurately weigh and transfer about 108.4 mg of working standard of Atorvastatin Calcium equivalent to 100 mg Atorvastatin into a 50 ml volumetric flask. Add 10 ml of Dimethyl sulphoxide &dissolve well. Further transfer about 34.5 mg of working standard of Amlodipine Besylate equivalent to 25 mg of Amlodipine into the above volumetric flask & add 30 ml solvent & shake for 20 minutes and dilute with solvent up to the mark and mix well. Transfer 10 ml of this solution into 50 ml volumetric flask & dilute with solvent up to the mark and mix well. Filter this solution with 0.45m membrane filter before injection. Sample            Weigh and powder 20 Tablets. Take powder containing equivalent to 100 mg of Atorvastatin & 25 mg of Amlodipine into 50 ml volumetric flask. Add 10 ml of Dimethyl sulphoxide & shake well. Add 30 ml solvent & shake for 20 minutes and dilute with solvent up to the mark and mix well. Transfer 10 ml of this solution into 50 ml volumetric flask & dilute with solvent up to the mark and mix well. Filter this solution with whatmann filter paper. Finally filter this solution with 0.45m membrane filter before injection. Condition       Waters Spherisorb 5mm ODS2   4.6 ´ 250mm./ UV at 254 nm / 11 minutes. Amlodipine Besylate: 4 minutes and Atorvastatin Calcium: 18 minutes (approximately). Gradient Progragram : Time(min.) Flow Rate Mobile Phase A Mobile Phase B 0.0 – 8.0 1.5 ml 40 60 8.0 –21.0 1.5 ml 50 50 21.01 – 30.0 1.5  ml 40 60