Sunday, February 3, 2013

Assay Method (M)





Assay Method of

Magnesium Hydroxide Tablet


Sample
Take a volume of test solution equivalent to 20 mg of magnesium hydroxide in a 250 ml conical flask. Add 100 ml of water and 10 ml of triethanolamine and stir. Add 5 ml of Ammonium - ammonium chloride buffer TS & 3 drops of eriochrome black indicator solution. Cool the solution between 3° & 4°C  by immersion of the flask in an ice bath. Then remove & titrate with 0.05 M Na2EDTA VS until a blue color produced. Each ml of 0.05 M Na2EDTA VS º 2.916 mg of Magnesium Hydroxide.

% of Mg(OH)2  =   No. of ml of 0.05 M Na2EDTA VS x F x 2.916 x 100 / Amount of Mg(OH)2  taken
Indicator
Dissolve 200 mg of eriochrome black T in a mixture of 15 ml of triethanolamine and 5 ml of dehydrated alcohol & mix.



Assay Method of

Mebendazole Syrup

Standard
10 mcg /ml of Mebendazole RS  in 0.1N Methanolic Hydrochloric acid TS. ( 0.001 % conc. )
Sample
Take suspension  equivalent to  100 mg of Mebendazole in 100 ml volumetric flask. Add 60 ml of 0.1N Methanolic Hydrochloric acid. and shake for 15 minutes. Make volume up to 100 ml with  0.1N Methanolic Hydrochloric acid . Filter.Dilute 1 ml of the filtrate to 100 ml with  0.1N Methanolic Hydrochloric acid .Measure the absorbance of the resulting solution at the maximum at 247 nm., taking 0.1N Methanolic Hydrochloric acid as reagent blank.


Assay Method of

Mebeverine  Hydrochloride Tablet


Sample
Weigh and powder 20 tablets. Heat a  quantity of powder  equivalent to  500 mg of Mebeverine  hydrochloride with 100 ml of 0.1M hydrochloric acid for 10 minutes on water bath , shaking occasionally. Cool , add sufficient 0.1M hydrochloric acid to produce 250 ml  . Mix and filter. To 10 ml of the filtrate add sufficient 0.1M hydrochloric acid to produce 100 ml and dilute 15 ml of the resulting solution to 100 ml with the same solvent. Measure the absorbance of the solution at 263 nm . Calculate the content of Mebeverine Hydrochloride taking 263 as the value of A (1%, 1cm ) at the maximum at 263 nm .
Calcula.
% of content  =  Absorbance of sample   x   100 / conc. in g /100 ml x 263



Assay Method of

Mebeverine  Hydrochloride SR Capsule

Standard
Accurately weigh and transfer about 75 mg of working standard of Mebeverine Hydrochloride into 100 ml volumetric flask. Add 70 ml of 0.1 M HCl and shake. Adjust the volume up to the mark with the same solvent. Dilute 1 ml of this solution to 50 ml volumetric flask with 0.1 M HCl.
Sample
Take 20 Capsules & determine the average content weight. Powder the pellets & transfer powdered sample equivalent to 75 mg of Mebeverine hydrochloride into a 100 ml volumetric flask. Add 70 ml of 0.1 M HCl and shake for 30 minutes. Adjust the volume up to the mark with the same solvent. Filter the solution. Take 1 ml of the filtrate into a 50 ml volumetric flask. Make the volume up to the mark with 0.1 M HCl & mix well.
Calcula.
Measure the absorbance of both Standard and sample solution in a UV-Visible Spectrophotometer having 1 cm cell at a wavelength of 263 nm using 0.1 M HCl as blank solution.


Assay Method of

Meclozine Hydrochloride Tablet

Buffer

Weigh about 3.45 gm of Monobasic Sodium Phosphate in 500 ml of water, and adjust pH to 4.0 with dilute Phosphoric acid.
Ratio   
Buffer: Methanol = 20: 80
Diluents
Methanol  and  Water
Standard 
Accurately weigh and transfer about 62.5 mg of Meclozine Hydrochloride Working Standard in a 100-ml volumetric flask. Add 50 ml of methanol and shake for 10 minutes and make up the volume with the water. Take 10 ml of this solution into a 50 ml volumetric flask and make up the volume with water Filter this solution with 0.45m membrane filters before injection.
Sample 
Weigh and powder 20 tablets. Take powder-containing equivalent to 62.5 mg of Meclozine Hydrochloride in a 100-ml volumetric flask. Add 50 ml of methanol and shake for 10 minutes and make up the volume with the water. Filter this solution with the whatman filter paper. From the filtrate take 10 ml of solution into a 50 ml volumetric flask and make up the volume with water Finally filter this solution with 0.45m membrane filters before injection.
Condition
Waters Spherisorb 10 mm C8  4.6 ´ 250mm./ 40°C  / UV at 230 nm / 15 minutes.


Assay Method of

Mecobalamin Tablet

M. Phase
Methanol: Water = 35: 65  
Ratio   
Buffer : Acetonitrile = 60 : 40
Diluents
Take 200 ml Methanol  in 500ml volumetric flask and made volume with water.
Standard 
Accurately weigh and transfer about 15 mg Mecobalamin INN  Working Standard in 50ml Amber Volumetric flask. Add 30 ml Diluents and shake for 10 minutes and make the volume up to   mark &mix well & Dilute 5ml to 50 ml with Diluents within 5 minutes.. Filter this solution with 0.25m membrane filters before injection within 5 minutes.
Sample 
Take powder-containing equivalent to 3 mg of Mecobalamin  in a 100 ml Amber volumetric flask. Add 70 ml of diluents & shake for 10 minutes. Dilute with Diluents up to the mark & filter with Whitman filter paper within 5 minutes. Filter This solution with 0.25 m membrane filter before injection within 5minutes.
Condition
Waters Spherisorb 10 mm ODS1 4.6 ´ 250mm./ 40º C / UV at 266 nm / 15 minutes.

Gradient Program
 
Serial No
Time
(minutes)
Flow
%of A Pump
(Solvent- Methanol)
% of  B Pump
(Solvent- Water)
1

1.0
35.0 %
65.0%
2
8.00
1.0
35.0 %
65.0 %
3
8.01
1.0
70.0 %
30.0 %
4
14.00
1.0
70.0 %
30.0 %
5
14.01
1.0
35.0 %
65.0%
6
20.00
1.0
35.0 %
65.0 %


Assay Method of

Mecobalamin  Injection

M. Phase
Methanol: Water = 65: 35  
Diluents
Distilled Water.
Standard 
Accurately weigh and transfer about 12.5 mg Mecobalamin INN Working Standard in 25-ml Amber Volumetric flask.
Add Diluents and shake for 15 minutes and make the volume up to mark & mix well. Filter this solution with 0.25m
membrane filters before injection.
Sample
Take 10 ml Injection in an amber Biker. Filter this solution with 0.25m membrane filters before injection.
Condition
Waters Spherisorb 10 mm ODS1 4.6 ´ 250mm./ 40º C / at 550 nm / 7 minutes / Inject. 100 ml.


Assay Method of

Mefenamic Acid / Tablet + Suspension

Standard 
Take 50 mg of Mefenamic Acid working standard in a 100 ml volumetric flask. Add 70 ml of diluent. Shake for 15 minutes. Add diluent to make volume 100 ml. Take 2 ml of the solution in a 50 ml volumetric flask. Make the volume up to mark (50 ml) with same diluent. Measure the absorbance at 350 nm taking above diluent as blank.
Sample
Weigh and powder 20 tablets. Take a quantity of the powder containing 50 mg of Mefenamic Acid in a 100 ml volumetric flask. Add 70 ml of diluent. Shake for 15 minutes. Add diluent to make volume 100 ml. Filter the solution. Take 2 ml of filtered solution in a 50 ml volumetric flask. Make the volume up to mark (50 ml) with same diluent. Measure the absorbance at 350 nm taking above diluent as blank.


Assay Method of

Meropenem Injection


Buffer
Take 1 ml triethylamine into a 1000 ml flask add 950 ml water then adjust pH 5.0 with phosphoric acid then volume 1000 ml with water.
M. Phase
Methanol: Buffer pH – 5.0 = 25: 75  
Diluents
Only  Buffer pH – 5.0
Standard 
Accurately weigh and transfer about 25 mg Meropenem for injection ( with sodium carbonate buffer ) USP Working
Standard in 50 ml Volumetric flask.Add Diluents and shake for 10 minutes and make the volume up to mark & mix well.
Filter this solution with 0.45mmembrane filters before injection.
Sample
Same as standard preparation.
Condition
Waters Spherisorb 10 mm ODS1 4.6 ´ 250mm./ at 300 nm / 10 minutes / Inject. 20 ml.


Assay Method of

Methyl Dopa Tablet

Glycin Buffer
Mix 42 g of sodium hydrogen carbonate and 50 g of potassium hydrogen carbonate with 180 ml of water and add a solution containing 37.5 g of glycine and 15 ml of 13.5M ammonia in 180 ml of water. Dilute to 500 ml with water and stir until solution is complete.
Standard 
Dissolve a 0.1 g of anhydrous methyldopa WS completely in sufficient 0.05M sulphuric acid to produce 100 ml and filter. To 5 ml of the filtrate add 2 ml of iron(II) sulphate-citrate solution, 8 ml of glycine buffer solution and sufficient water to produce 100 ml.
Sample
Dissolve a quantity of the powder containing the equivalent of 0.1 g of anhydrous methyldopa as completely as possible in sufficient 0.05M sulphuric acid to produce 100 ml and filter. To 5 ml of the filtrate add 2 ml of iron(II) sulphate-citrate solution, 8 ml of glycine buffer solution and sufficient water to produce 100 ml. Measure the absorbance of the resulting solution at the maximum at 545 nm using
Blank
Dilute 2 ml of iron(II) sulphate-citrate solution, 8 ml of glycine buffer solution to 100 ml with water.


Assay Method of

  Methyl Ergonovine Maleate Tablet
                             .
Sample 
Weigh & powder 20 tablets . Dissolve a quantity of powder equivalent to 2 mg of Methyl Ergometrine Maleate in 50 ml of  a 1 % w/v solution of tartaric acid. Mix and filter . To 3 ml add 6 ml of dimethylaminobenzaldehyde solution , mix, cool in running water for five minutes, add sufficient dimethylaminobenzaldehyde solution to produce 10 ml ( Solution A ). At the same time prepare two similar solutions, one with Methyl Ergometrine Maleate RS (Solution B ) and the other without  the substance being examined. ( Solution C ) .
Measure the absorbance  of Solution A & B at 590 nm, using solution C as the blank,
               

                                              Assay Method of             

                                               Methyl Ergonovine Maleate Injection
                             .
Standard
Dissolve 25 mg of Methyl Ergonovine Maleate in 50 ml of water . Dilute 4 ml to 50 ml with water .
Sample 
Dilute a quantity of  sample equivalent to 2 mg of Methyl Ergonovine Maleate ( 10 ml ) in 50 ml of  water.  Mix . To 3 ml add 6 ml of dimethylaminobenzaldehyde solution ( # solution ) , mix, cool in running water for five minutes, add sufficient di-methylaminobenzaldehyde solution to produce 10 ml ( Solution A ). At the same time prepare two similar solutions, one with Methyl Ergonovine Maleate WS (Solution B ) and the other without  the substance being examined. ( Solution C ) .Measure the absorbance  of Solution A & B at 590 nm, using solution C as the blank,
 # solution
Dissolve 0.125 mg of 4-Dimethyl benzaldehyde in a cooled mixture of 35 ml water and 65 ml of sulphuric acid. Add 0.1ml of 5 % w/v solution of  iron (III) chloride hexahydrate  .Allow to stand for 24 hours, protect from light , before use. Discard the solution if any yellow colour appears.


Assay Method of

Metoclopramide HCl  Syrup

Reagent 
01. 1 % w/v Sodium Nitrite 02. 5 % w/v of Ammonium Suphamate  03. 0.5 % w/v solution of N-(2-naphthyl) ethylenediamine dihydrochloride in 1M Hydrochloric acid  and All reagents should be freshly prepared .
System 
Protect the solutions from light . Dilute a quantity of sample solution containing 10 mg of Anhydrous Metoclopramide HCl  to 100 ml with water. To 5 ml of this solution add 5 ml of M hydrochloric acid , mix . Add 5ml of sodium nitrite solution , mix and allow to stand for 20 minutes . Add  5 ml of  Ammonium sulphamate solution , mix and allow to stand for 25 minutes. Add 5 ml of  N-(2-naphthyl) ethylenediamine dihydrochloride solution  , mix and dilute with water to 100 ml . Allow to stand for 5 minutes.Measure the absorbance of the resulting solution at the maximum at 534 nm , using water in the reference cell. Calculate the content of  Metoclopramide  Hydrochloride from the absorbance  obtained by repeating the procedure using 5 ml of 0.01 % w/v solution of Metoclopramide Hydrochloride RS in place of the solution being examined.
     

                               Assay Method of                                     

Metformin Hydrochloride Tablet
                             .
Sample 
Shake a quantity of the powder containing 0.1 g of Metformin Hydrochloride with 70 ml of water for 15 minutes, dilute to 100 ml with water and filter, discarding the first 20 ml. Dilute 10 ml of the filtrate to 100 ml with water and dilute 10 ml of the resulting solution to 100 ml with water. Measure the absorbance of the resulting solution at the maximum at 232 nm using water as blank.
Calculation
 % of Metformin Hydrochloride =  Absorbance of Sample ´ 100 / Conc. Of spl. ´ 798                                              


     Assay Method of

     Metronidazole Tablet
                              .
Sample 
Take powder equivalent to 50 mg of metronidazole in a 100ml volumetric flask. Add 60 ml of 0.1M hydrochloric acid.  Shake for 15 minutes. Make the volume up to 100ml with 0.1M hydrochloric acid. Mix and filter. Dilute 1 ml of the filtrate to 50 ml with 0.1M hydrochloric acid. Measure the absorbance of the resulting solution at 277 nm using 0.1M hydrochloric acid as blank.
Calculation
 % of Metronidazole =  Absorbance of Sample ´ 100 / Conc. Of spl. ´ 375                                              
                       

Assay Method of

Metronidazole Suspension
                              .
Sample 
Take  suspension equivalent to 100 mg of metronidazole  in a 250 ml separating funnel. Rinse the pipette with 10 ml of water. Extract two times each with 40 ml of chloroform. 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   equivalent to 17.12 mg of  metronidazole.
Calculation
% of content   =   No. of ml of 0.1N perchloric acid VS x F x 17.12 x 100 / 100 mg


Assay Method of

Miconazole / Gel  or  Miconazole Nitrate + Hydrocortisone Cream


Buffer
For 1 liter buffer take about 6 gm of Ammonium Acetate in the mixer of 550 ml distilled water & 450 ml methanol, then shake for 5 minutes for properly dissolve finally filter & sonicate this buffer solution.
Ratio   
Buffer 45 %  :  Acetonitrile 55 %
Diluents
Methanol
Standard 
Dissolve about 25 mg of Hydrocortisone ( 1 mg Hydrocortisone ≡ 1.116 mg Hydrocortisone Acetate ) WS & 50 mg of Miconazole Nitrate WS in a 50 ml volumetric flask then add 35 ml methanol and shake for 25 minutes and dilute with methanol up to mark 50 ml and mix well finally filter this solution with 0.20 micron membrane filter before injection.
Sample 
Take about 2.5 gm of sample cream equivalent to 25 mg of Hydrocortisone & 50 mg of Miconazole Nitrate in a 50 ml volumetric flask then add 35 ml methanol and shake for 25 minutes and dilute with methanol up to mark 50 ml and mix well, filter this solution with whattman filter paper & finally filter this solution with 0.20 micron membrane filter before injection.
Condition
Waters Spherisorb 5 mm ODS2 4.6 ´ 250mm./ 50°C  / UV at 235 nm 3 minutes for Hydrocortisone & 7 minutes for Miconazole Nitrate ( Approximately)
                                                           

Assay Method of

 Midazolam Tablet
                              .
System 
Take standard & sample as concentration = 0.001%
Solvent = 0.1 M HCl
Wave Length = 258 nm


Assay Method of

Mizolastine Tablet


Buffer

Transfer accurately weighed about 1.70 gm of potassium Dihydrogen orthophosphate (KH2PO4)  in 450 ml purified
water, mix well. Adjust PH to 3.5 ± 0.05 with orthophosphoric acid and dilute to 500 ml with distilled water. Filter
through a 0.45mm membrane filter.
Ratio   
Buffer : Acetonitrile in the proportion of  70 : 30
Standard 
                                    Weigh accurately about 10mg Mizolastine WS and transfer in a 25ml volumetric flask. Add 15 ml of mobile phase. Shake for 25min. Then add same diluent to make 25 ml. Mix and filter this solution with 0.45mm membrane filter before injection.
Sample 
Take 20 tablets in a mortar and finely crushes with passel. Take accurately weight of the Tablet powdered sample
containing about 10mg Mizolastine and transfer in a 25 ml volumetric flask. Add 15ml of  mobile phase. Shake for
25min and add same diluent to make 25 ml. Mix and filter with whatman filter paper. Filter this resultant solution
with 0.45 mm membrane filter before injection .
Condition
ODS1  , 5 m 4.6 x 250 mm / UV at 212 nm/10 minutes

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