Analytical Method of Medicinal Product Starting Generic Name with word “A”

Solvent

Methanol

Standard

50 mg —-® 100 ml —-® take 2 ml —-® 50 ml  (Shake 30 minutes)

Sample

Same as standard. Take absorbance at 276 nm.

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.

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, 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

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.)

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.

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.

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

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

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.

Solvent

Methanol

Standard

25 mg —-® 100 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.

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

Accetonitrile : Buffer º 40 : 60

Diluents

Accetonitrile : 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).

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).

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

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.

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.

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 theConical 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.

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

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

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.

M. Phase

34.84 g/L solution of  Dipotassiums 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.