Wednesday, February 13, 2013

Assay Method (S)

Assay Method of
 Salbutamol  Syrup

Reagent 
1. 1M Sodium carbonate solution in water 2. 0.2 % w/v P- aminophenol in ethanol
Standard 
Take Salbutamol sulphate equivalent to 100 mg of salbutamol in a 100 ml volumetric flask. Add 60 ml of water and shake to dissolve .  Dilute to 100 ml with water. Dilute 2 ml  of the resulting solution to 25 ml with water.
Concentration 0.008 %
Sample 
Take 20 ml of syrup equivalent to 8 mg of  Salbutamol in a  100 ml volumetric flask. Dilute with water  to volume and mix. 1 mg  of Salbutamol  equivalent to 1.2049 mg of Salbutamol sulphate
System
Take 3 ml of  sample and standard solution each in two separate 25 ml of volumetric flask. Add 3 ml of  1M sodium carbonate solution and 0.2 ml of P-aminophenol solution to each flask. Dilute to 10 ml with water. Keep at rest for 25 minutes. Measure the absorbance at 635 nm taking water as blank.

Assay Method of
 Sertraline HCl Tablet

Sample 
Take the weigh of   20 tablets & determine the average weight. Then crush them into fine powder. Take powdered sample equivalent to 100 mg of Sertraline Hydrochloride and dissolve in 30 ml Glacial Acetic Acid. Add 10 ml of 6% Mercuric Acetate solution. Carry out a potentiometric titration with 0.1 N Perchloric acid to determine the end point using α-napthyl benzene as an indicator. Perform blank titration If necessary.
1 ml 0.1N Perchloric acid VS ≡ 34.27 mg of Sertraline HCl
Result
Calculation : % of Sertraline   = V x f x C x 100 / W

Assay Method of
 Sildenafil Citrate Tablet

Method – 01/HPLC
Buffer

2.720 gm of Potashium Di Hydrogen Phosphate in 1000 ml volumetric Flask with distilled water, adjust pH 6.5 ± 0.5 with 2N Sodium Hydroxide. filter & sonicate this buffer solution.
Mobile Phase

70% Methanol : 30% Buffer
Diluent  
Methanol
Standard 
10 mg ---® 100 ml
Sample 
10 mg ---® 100 ml
Condition
XTerra Column 4.6 x 250 mm / 400 C/ 292 nm / 1 ml per minute / 10 micro liter / 6 minutes approx

Method – 02/UV
Solvent
Method - 01
Standard 
10 mg ---® 100 ml ---® take 10 ml ---® 50 ml (Shake 10 minutes)
Sample 
Same as Standard. Take absorbance at 292 nanometer.

Assay Method of
 Silver Sulphadiazine Cream

Ratio   
Water : Acetonitrile  : Phosphoric Acid  : : 90 : 9.9 : 0.1
Diluent  
Transfer 100 ml of Ammonia solution (25% concentrate) to a 1 liter volumetric flask & dilute with water to volume & mix.
Standard 
Weigh and transfer about 25 mg  working standard of Silver Sulphadiazine into a 50 ml volumetric flask.Add 30 ml solvent & shake for 20 minutes and dilute to 50 ml with the same solvent and mix well. Transfer 5 ml of this solution in a 50 ml volumetric flask & volume with solvent up to the mark & mix well.  Filter this  solution with 0.45 mic membrane filter before injection.
Sample 
Transfer an accurately weighed quantity of Neozine Cream equivalent to 25 mg of Silver Sulphadiazin in a separating funnel. Add 50 ml of solvent & shake for 10 minutes. Then add 50 ml of chloroform in to the separating funnel & shake for another 10 minutes. Wait for 30 minutes to separate the layer completely, discard the lower layer & collect the upper layer into a 250 ml stopper conical flask & evaporate to remove the chloroform. Cool the solution & transfer it into a 50 ml volumetric flask & volume it with the same solvent. Transfer 5 ml the solution into 50 ml volumetric flask & dilute with the solvent up to the mark. Then filter the this solution with whatmann filter paper & finally filter this solution with 0.45 m m  membrane filter before injection.
Condition
ODS2, 10mm  4.6 x 250 mm / 254 nm / 10 minutes / 7 minutes (approximately)
                                                                                                           
Assay Method of
Simethicon Solution & Tablet

Standard
Take 134 mg of Simethicone Compound in a 50 ml volumetric flask. Add 30 ml of 1 M Sodium Hydroxide and shake for 10 minutes and dilute with 1 M Sodium Hydroxide up to the mark Add this solution into the separating funnel which is pre-washed with 15 ml of 1 M Sodium Hydroxide after adding 67 ml of toluene. Shake this solution for 5 minutes. Allow the mixture to stand for 20 minutes, centrifuge the upper layer and dry with anhydrous sodium sulphate. Measure the absorbance of Simethicone at 1261 cm-1 By  IR Machine.
Sample
Accurately take 2 ml of Flatulex Drops equivalent to 134 mg Simethicone into a 50 ml volumetric flask. Add 30 ml of 1 M Sodium Hydroxide and shake for 10 minutes and dilute with 1 M Sodium Hydroxide up to the mark Add this solution into the separating funnel which is pre-washed with 15 ml of 1 M Sodium Hydroxide after adding 67 ml of toluene. Shake this solution for 5 minutes. Allow the mixture to stand for 20 minutes, centrifuge the upper layer and dry with anhydrous sodium sulphate. Measure the absorbance of Simethicone at 1261 cm-1 By  IR Machine.
        
Assay Method of
 Sulphamethoxazole + Trimethoprim Syrup

Sulphamethoxazole
Dissolve suspension equivalent to 200 mg of Sulphamethoxazole in 60 ml of water and 10 ml of hydrochloric acid. Add 3 g of potassium bromide, cool in ice and titrate slowly with 0.1M Sodium Nitrite VS, stirring constantly and determine the end point electrochemically. Each ml of 0.1M Sodium Nitrite VS is equivalent to 25.33 mg of Sulphamethoxazole.

% of SMZ = No. of ml of VS x F x 25.33 X 100 / Amount of Sulphamethoxazole taken
Trimethoprim
Method I :
To a amount of suspension  containing 50 mg of Trimethoprim add 30 ml of 0.1M Sodium Hydroxide and extract with  two 50 ml quantities of  chloroform, washing each extract with the same two 10 ml quantities of 0.1M Sodium Hydroxide. Combine the chloroform extracts and extract with three 50 ml quantities of  1M Acetic Acid. Dilute the aqueous extracts to 250 ml with 1M Acetic Acid. To 10 ml of the resulting solution add 10 ml of 1M Acetic Acid and sufficient water to produce 100 ml .Measure the absorbance of the resulting solution at the maximum at 271 nm , taking water as blank.Calculate  the content of Trimethoprim  taking 204 as the value of A ( 1%, 1 cm ) at maximum at 271 nm.

% of Trimethoprim  = Absorbance     x   100 / conc. In gm per 100 ml  x  204

Method II :
Take  sample equivalent to 80 mg of Trimethoprim  in a 250 ml separating funnel. Add 30ml of 0.1N sodium hydroxide. Extract two times, each time with 50ml of chloroform. Wash the total extract with 20 ml of water. Collect the extracts in a 400 ml beaker and evaporate the extracts to dryness on a water bath. Dissolve the residue in 30 ml of anhydrous glacial acetic acid. Titrate with 0.1N perchloric acid VS using  naptholbenzein solution as indicator. 1 ml of 0.1N perchloric acid VS   is º 29.03 mg of  Trimethoprim.

% of content  =   No. of ml of VS x F x 29.03 x 100 / Amount of Trimethoprim taken

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