Sunday, February 3, 2013

Assay Method (G)







Assay Method of
Gliclazide Tablet

Standard 
Take 25 mg of working standard of Gliclazide in a 100 ml volumetric flask. Add 70 ml of Methanol and shake for 10 minutes.  Adjust the volume to100 ml with Methanol. Take 2 ml of the solution in a 50 ml volumetric flask. Make the volume up to the mark with Methanol. Take absorbance at 228 nm.Use Methanol as blank.
Sample 
Weigh and make fine powder of 20 tablets. Take an accurately weighed portion of powder equivalent to 25 mg of Gliclazide in a 100 ml volumetric flask. Add 70 ml of Methanol and shake for 30 minutes.  Adjust the volume to100 ml with Methanol & mix.  Filter the solution by Whatman filter paper. Take 2 ml of the filtrate in a 50 ml volumetric flask. Make the volume up to the mark with Methanol. Take absorbance at 228 nm.Use Methanol as blank


Assay Method of
Glimepiride Tablet

Solvent
Methanol
Standard 
25 mg ----® 100 ml ----® Take 2 ml ----® 50 ml
Sample 
Same as standard. wavelength 228 nm

Guiphenesin + Pseudoephedrine HCl + Triprolidine HCl Syrup

Guiphenesin
Standard preparation :
Dissolve 100 mg of Guaiphenesin RS in 100 ml chloroform. Dilute  5 ml to 100 ml with chloroform. Conc. 0.005 %
Assay preparation :
To 5 ml syrup add 10 ml of saturated sodium bicarbonate solution and 10 ml of water in a separating funnel. Extract four time each with 20 ml of chloroform , filtering each extract through chloroform washed cotton in a second separator. Wash the combined chloroform extract with 5 ml of N hydrochloric acid and filter through cotton ,washed with chloroform in to a 100 ml  volumetric flask.. Make volume up to 100 ml with chloroform.Dilute 5 ml to 100 ml with chloroform.Conc. 0.005 % .Measure the absorbance  at 276 nm taking chloroform as blank
Pseudo. HCl
Standard preparation :
Dissolve 30 mg of Pseudoephedrine HCl RS in 50 ml water. Dilute  5 ml to 100 ml with water.Conc. 0.003 %
Sample preparation      :
Dilute 5 ml of syrup to 50 ml with water. Mix . Dilute 5 ml to 100 ml with water .
Assay procedure :
Take 5 ml standard preparation , 5 ml sample preparation , and 5 ml water in three separating funnel. Add 1ml saturated sodium carbonate solution , 2 ml metaperiodate solution ( 1 in 50 ) in each funnel, allow to stand for 10 minutes. Add 20 ml of n – Hexane to each  three funnel. Shake for 30 secs. Allow to separate , collect  the n-Hexane layer in three dry test tubes.
Measure the absorbance at 241 nm taking n-Hexane as blank,Conc. 0.003 %
Triproli.HCl
Standard preparation :
Dissolve 25 mg of Triprolidine HCl RS in 200 ml HCl ( 1 in 200 ml ). Dilute  5 ml to 100 ml with the same solvent.
Conc. 0.00125 %
Assay procedure :
Take 5 ml  syrup in a separating funnel , add 10 ml of Sodium Hydroxide ( 1 in 10 ml ) and extract with 2 x 50 ml n-Hexane. Combine the extract in a second separator and wash with 2 x 10 ml of Sodium Hydroxide solution ( 1 in 250 ml ). Discard the washing. Extract the n-Hexane solution with 2 x 40 ml dil. HCl ( 2 in 200 ml ).Collect the acid layer in 100 ml volumetric flask & make the volume 100 ml with dilute HCl.( 1 in 100 ml ), Measure the absorbance at  290 nm in taking dilute HCl in the reference cell.


Assay Method of
Gemifloxacin Tablet

Mobile Phase
Acetonitrile: Buffer (50:50)             
Diluents
Acetonitrile: Buffer (30:70)                
Buffer
Dissolve 3.12gm of Sodium dihydrogen phosphate dihydrate in 1000 ml of distilled water and adjust the pH to 2.0 ± 0.05 with dilute ortho-phosphoric acid. Filter through 0.45mm membrane filter and degas.
Standard 
Weigh accurately and transfer about 31.172 mg Gemifloxacin Mesylate standard equivalent to 25 mg Gemifloxacin into a 50 ml volumetric flask, add 25 ml diluent to  dissolve and make up to the mark with diluent and then dilute 5.0 ml of above solution to 50 ml with diluent. Mix and filter this solution with 0.45mm membrane filter before injection.
Sample 
Weigh 20 tablets and determine the average weight. Then crush these tablets into fine powder using mortar and pastle. Weigh accurately and transfer about 43 mg powdered sample of tablets equivalent to 25 mg Gemifloxacin into a 50 ml volumetric flask. Add 25 ml of diluents to dissolve & shake for 10 minutes. Then add sufficient diluents to make 50ml and then dilute 5.0 ml of above solution to 50 ml with diluent. Mix and Filter with whatman filter paper. Filter this resultant solution with 0.45 mm membrane filter before injection.
Condition
Xterraâ RP 18 Column (4.6 mm x 250mm), UV at 274 nm , 1ml/minute, 2.5 minutes (approximately)


Assay Method of
Haloperidol Tablet

Mobile Phase
Mix filtered 1% Ammonium Acetate solution & Acetonitrile in the ratio of 45:55(v/v) . Degas the mixture for 30 minutes.
Diluents
Mis filtered1% Ammonium Acetate solution & Acetonitrile in the ratio of 50:50(v/v). Degas the mixture.
Standard 
Weigh accurately 20 mg of Haloperidol working standard in a 100 ml volumetric flask.Add 50 ml of Diluting solvent and shake for 30 min. Make up to volume with the diluting solvent. Filter the std with 0.2µ filter paper. concentration will be0.02%.
Sample 
Weight accurately 10 mg Haloperidol containing equivalent tablet powder in a 50 ml volumetric flask. Add 30 ml of dilution solvent and shake for 30 minutes. Make up to volume with the diluting solvent. Filter the spl with Whatman filter paper first then with 0.2µ filter. Concentration will be 0.02%
Condition
Supilcosil DiscoveryTM C18 Column (25cmX4.6mm) & 5µm. ID 504971/ UV at 247 nm


Assay Method of
Hydrocortisone Injection

Sample 
Dissolve the contents of  a sealed container in sufficient water to produce  a solution containing the equivalent of 0.001 % w/v solution  of hydrocortisone. Measure the absorbance of the resulting solution at the maximum  at 248 nm .
Calculate the content of hydrocortisone taking 449 as the value of  a (1% , 1 cm ) at the maximum at 248 nm . Repeat the the procedure with  a further nine sealed containers  and calculate the average content of hydrocortisone per container from the ten individual results thus obtained.
% of content = Absorbance  x 100 / conc. in g per 100 ml x 449  


Assay Method of
Hydroxazine HCl Tablet

Buffer

Transfer accurately weighed about 6.8 gm of potassium Dihydrogen orthophosphate (KH2PO4) in 950 ml distilled water, mix well. Adjust pH to 2.7 ± 0.05 with orthophosphoric acid and dilute to 1 liter with distilled water. Filter through a 0.45mm membrane filter.
Ratio   
Acetonitrile : Buffer in the proportion of  70 : 30
Diluents
Methanol : Buffer in the proportion of  50 : 50
Standard 
                                    Weigh accurately about 20mg Hydroxyzine HCl WS and transfer in a 50 ml volumetric flask. Add 30 ml of diluents Shake for 10 min. Then add sufficient diluents to make 50ml. Mix and filter this solution with 0.45mm membrane filter before injection.
Sample 
Take the weight of 20 tablets and determine the average weight. Then crush the tablets in to fine powder.    Take the
powdered sample equivalent to 20 mg Hydroxyzine HCl and transfer in a 50 ml volumetric flask. Add 30 ml of
diluents & Shake for 10 minutes. Then add sufficient diluents to make 50ml. Mix and Filter with whatman filter
paper. Filter this resultant solution with 0.45 mm membrane filter before injection.
Condition
ODS1  , 10 mm 4.6 x 250 mm / UV at 232 nm / 8 minutes


                                            Assay Method of
                                 Hyoscine Butylbromide Tablet

Mobile Phase A

Dissolve 7.0 gm of Potassium Dihydrogen Phosphate with 900 ml  distilled water into 1000 ml volumetric flask, adjust the pH 2.0 ± 0.05 with Phosphoric acid  & make volume with distilled water up to 1000  ml.
Mobile Phase B

Dissolve 5.8 gm of Sodium Lauryl Sulphate with 400 ml  distilled water into 1000 ml volumetric flask, Then add 400 ml Acetonitrile and adjust the pH 2.0 ± 0.05 with Phosphoric acid  & make volume with distilled water up to 1000  ml.
Ratio   
Mobile Phase A ( 70 % ) : Mobile Phase B ( 30 % )
Diluent  
0.01M Hydrochloric Acid
Standard 
                                Dissolve about 20 mg of Hyoscine Butylbromide WS in 10 ml of water and shake for 10 minutes. Then volume 100 ml with diluents. Filter this solution with 0.45 m membrane filter before injection.
Sample 
Take a quantity of the powdered sample containing about 20 mg of Hyoscine Butylbromide in 10 ml of water and shake for 10 minutes. Then add about 50 ml diluents and shake for 15 minutes .Dilute to 100ml diluents. Filter to obtain a clear filtrate using by Whattman filter paper & next filter this solution with 0.45 m membrane filter before injection.
Condition
Waters Spherisorb®  S10 C8 4.6 x 250 mm / 400 C/ UV at 210 nm/10 minutes
   


                                      Assay Method of
                          Hyoscine Butylbromide Injection

Sample 
Dilute 200 mg of Hyoscine butyl bromide with 50 ml of water. Acidify  with 2 M nitric acid . Titrate with 0.1M silver nitrate , determining the end –point potentiometrically using a silver indicator electrode and a silver-silver chloride reference electrode.  (Electrode DM –141 , Instrument Mettler DL 40 RC Memotitrator )
Calculation
No. of ml of 0.1M silver nitrite VS x F x 44.04 x 100 / 200 mg

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