Saturday, February 2, 2013

Assay Method (C)










Calcium Carbonate / Tablet

Sample
Take the amount of powder equivalent to about 40 mg of calcium transfer to conical flask (250ml). Add 10ml of distilled water then heat on water bath about 15-20 minute then cool it.  Dissolve the residue by adding sufficient (1-2 ml) 3N hydrochloric acid drop-wise, to achieve complete solution. Then dilute with water to 150ml, add 15ml of 1N sodium hydroxide and 3-5 mg of Hydroxy naphthol blue as indicator and titrate with 0.05M Edetatedisodium VS until the solution is deep blue.1 ml 0.05M Edetate disodium  º 2.004 mg of Calcium
                          
%  of content = V1 ´ F ´ 2.004 ´ 100 / W


Calcium Lactate / Tablet

Sample
Dissolve a quantity of the powder containing 0.300 g of Calcium Lactate Pentahydrate in 50 ml of water and titrate with 0.05 M Disodium Edetate VS to within a few milliliters of the expected end point. Add 8 ml of 5M Sodium Hydroxide  and 0.05 g of Solochrome dark blue mixture and continue the titration until the colour changes from pink to full blue.

Each ml of 0.05 M Disodium Edetate VS º 15.41 mg of Calcium lactate pentahydrate .
    
% of content  = Required volume of  VS x F x 15.41x 100 / Weight of Calcium Lactate Pentahydrate                   


Calcium Pantothenate / Injection

Sample
Take sample equivalent to 200 mg of Calcium Pantothenate in a 250 ml titration flask. Add 200 ml water, 4 ml 10 M sodium hydroxide & 15 mg of calcon carboxylic acid triturate. Titrate with 0.05 M Na2EDTA vs until the color changes from violet to full blue. Each ml of 0.05 M Na2EDTA VS is equivalent to 23.825 mg of Calcium Pantothenate .
  
Calculation: V x F x 23.825 x 100 / weight of Calcium Pantothenate in mg. 


Captopril / Tablet

Solvent
0.1M Sodium Hydroxide
Standard 
50 mg ----® 100 ml ----® take 2 ml ----® 50 ml  (Shake 25 minutes)
Sample  
Same as standard. Measure the absorbance at 238 nm Use 0.1M Sodium Hydroxide as blank.


Carbamazepine / Tablet

Solvent
Ethanol
Standard 
50 mg ----® 50 ml ----® take 5 ml ----® 50 ml  (Shake 10 minutes)
Sample 
Same as standard. Measure the absorbance at 285 nm Use 0.1M Sodium Hydroxide as blank.


Carvidilol / Tablet

Buffer
Dissolve 2.723 gm of Potassium dihydrogen orthophosphate with 950 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.
Ratio   
Buffer 65%  :  Acetonitrile 35%  
Solvent
1st Dilution – Acetonitrile & 2nd  Dilution – Acetonitrile 60% : Water 40%
Standard 
25 mg ----® 50 ml ----® take 2 ml ----® 25 ml  (Shake 20 minutes)
Sample  
Same as standard.
Condition
C8 5.0 mm 4.6 x 250 mm / 55°C / UV at 240 nm / 8 minutes
                    

Cefaclor / Powder for Suspension

Solvent
Water
Standard 
100 mg ----® 100 ml ----® take 2 ml ----® 100 ml  (Shake 10 minutes)
Sample 
Same as standard. Measure the absorbance at 265 nm Use water as blank.


Cefepime HCl / Injection

Buffer
Weigh 2.88 gm of Hexane –1 Sulphonic acid sodium salt.& transfer to a 1000 ml volumetric flask. Add approx. 500 ml distilled water to dissolve completely & volume up to the mark with distilled water &mix well. Adjust pH 4.0 with dilute glacial acetic acid or Potassium Hydroxide (if required).
Ratio   
Buffer ( 90% ) : Acetonitrile ( 10% )    
Solvent
Mobile Phase
Standard 
25 mg ----® 50 ml (Shake 20 minutes)
Sample 
Same as standard.
Condition
Waters Spherisorb  S10 C8, 4.6 x 250 mm / UV at 254 nm / 15 minutes



Cefixime / Capsule + Powder for Suspension

Solvent
Methanol
Standard 
50 mg ----® 100 ml ----® take 2 ml ----® 100 ml  (Shake 20 minutes)
Sample 
Same as standard. Measure the absorbance at 290 nm Use methanol as blank.


Ceftazidim Pentahydrate / Injection

Buffer pH = 7.0
Dissolve 42.59 g anhydrous dibasic sodium phosphate and 27.22 g of monobasic potassium phosphate in water to make 1000 ml of solution.
Ratio   
Mix 70 ml of acetonitrile and 100 ml of pH 7 buffer and dilute with water  to obtain 1000 ml of solution. Filter this solution with 0.45 m membrane filter and degas.
Solvent
Mobile Phase
Standard 
Ceftazidime Pentahydrate (with buffer) working standard equivalent to about 50 mg of Ceftazidime 50 mg ----® 50 ml ----® take 5 ml ----® 50 ml  (Shake 20 minutes)
Sample 
Same as standard.
Condition
Waters Spherisorb ODS1 10m, 4.6 x 250 mm / UV at 254 nm / 40m / 1.5 ml per minutes / 8 minutes


Ceftriaxone / Injection

Sample 
Determine the weight of the contents of 10 containers as described in the test for uniformity of weight. Dissolve a quantity of the mixed contents of the 10 containers containing the equivalent of 0.05 g of ceftriaxone in sufficient water to produce 100 ml. Dilute 2 ml to 100 ml with water.
Prepared standard solution in the same way. Measure the absorbance of the resulting solutions at 242 nm using water as blank . Concentration 0.001 %


Cefuroxime Axetil / Tablet & Powder for Suspension

Solvent
Methanol
Standard 
36 mg Cefuroxime Axetil equivalent 30 mg Cefuroxime ----® 50 ml ----® take 1 ml ----® 50 ml  (Shake 15 minutes)
Sample 
Same as standard. Measure the absorbance at 277 nm Use methanol as blank.


Cephalexin / Capsule

Sample 
Weigh and powder 20 capsules. Take powder  equivalent to  50 mg of Anhydrous Cephalexin in 100 ml volumetric flask. Add 60 ml of  water , and shake for 25 minutes. Make the volume up to 100 ml with water. Mix and filter. Dilute 2 ml of the filtrate to 50 ml with water.  Measure the absorbance of the resulting  solution  at the maximum 262 nm,Calculate the content of  Cephalexin taking 232 as the value of A ( 1%, cm ) at 262 nm.

% of content  =  Absorbance of sample   x   100 / Conc. in g / 100 ml   x   232
         

 Cephradine / Injection

Sample 
Determine the weight of the contents of 10 containers. Dissolve a quantity of the mixed contents of the 10 containers containing the equivalent of 0.1 g of cephradine in sufficient water to produce 100 ml. Dilute 1 ml to 50 ml with 0.1 M hydrochloric acid. Prepare a standard solution in the same concentration by using cephradine Reference Standard. Measure the absorbances of the resulting solutions at the maximum at 257 nm using 0.1 M hydrochloric acid as blank. Concentration 0.002 % w/v


Cephradine / Capsule & PFS

Solvent
Water
Standard 
100 mg ----® 100 ml ----® take 2 ml ----® 100 ml  (Shake 15 minutes)
Sample 
Same as standard. Measure the absorbance at 261 nm Use water as blank.


Cetirizine Hydrochloride / Tablet

Solvent
0.1 M HCl
Standard 
50 mg ----® 100 ml ----® take 2 ml ----® 50 ml  (Shake 20 minutes)
Sample 
Same as standard. Measure the absorbance at 231 nm Use 0.1 M HCl as blank.


Cetirizine Hydrochloride / Syrup

Mobile Phase          
A  = Methanol  :  Water  = 20  : 40 and  B = Acetonitrile                      
Solvent
Methanol : Water :  Acetonitrile  = 20  :  40  :  40 
Ratio
Binary gradient system  = A : B = 70  :  30
Standard 
Dissolve 25 of Cetrizine Hydrochloride WS in 50 ml of solvent . Dilute 5 ml to 25 ml with the same solvent.
Sample 
Dilute 5 ml sample to 25 ml with the solvent.
Condition
Supilcosil Discovery   TM C 18 Column / 230 nm / 2.3 ml per minute / 40 µl    


Cinchocaine HCl

Solvent
1M HCl
Standard 
30 mg ----® 50 ml ----® take 1 ml ----® 50 ml  (Shake 15 minutes)
Sample 
Same as standard. Measure the absorbance at 254 nm Use 1M HCl as blank.


Cinnarizine / Tablet

Solvent
0.1M HCl
Standard 
50 mg ----® 100 ml ----® take 1 ml ----® 50 ml  (Shake 25 minutes)
Sample 
Same as standard. Measure the absorbance at 254 nm Use 0.1M HCl as blank.


Ciprofloxacin   / PFS

Buffer
Add 1 ml triethylamine in 500 ml of water and adjust the PH  3.0( ±0.05) with dilute phosphoric acid.
Ratio   
Acetonitrile : Buffer in the proportion of  35 : 65
Solvent
1st Dilution – 0.1M HCl & 2nd  Dilution – Water
Standard 
                                    55.5 mg Ciprofloxacin Hydrochloride  working  standard  equivalent to Ciprofloxacin 50 mg ----® 50 ml ----® take 2.5 ml ----® 50 ml  (Shake 10 minutes)
Sample 
Same as standard.
Condition
Waters Spherissorb ODS1  10 mm 4.6 ´ 250mm./ UV at 278 nm / 8 minutes

Ciprofloxacin   / Tablet

Sample 
Dissolve powdered sample equivalent to 100 mg Ciprofloxacin in previously neutralized mixture of 60 ml glacial acetic acid and 5% Mercury (II) Acetate (50 ml + 10 ml) using crystal violet solution as indicator. Titrate with 0.1 M Perchloric acid and determine the end point.Each ml of 0.1M perchloric acid VS is equivalent to 33.14 mg of ciprofloxacin          
% of content =   V x F x 33.14 x 100 / W


Ciprofloxacin   / Tablet
Buffer
Add 1 ml triethylamine in 500 ml of water and adjust the PH  3.0( ±0.05) with dilute phosphoric acid.
Ratio   
Acetonitrile : Buffer in the proportion of  35 : 65
Diluents
Acetonitrile : Buffer in the proportion of  35 : 65
Standard 
                                    Accurately  weigh and  transfer about  55.5 mg Ciprofloxacin Hydrochloride  working  standard  equivalent to 50 mg of Ciprofloxacin in a 50 ml volumetric flask .Add diluents and shake for 25 minutes and make the volume up to the mark with the same solvent. Dilute 25 ml of this solution to 100ml with diluents. Filter this solution with 0.20 m membrane filter before injection.  
Sample 
Take an accurately powder equivalent to 250 mg of ciprofloxacin in 100 ml volumetric flask. Add 70 ml of diluents and shake for 30 minutes & sonicate for 30 minutes.. Adjust the volume to100 ml with diluents. Filter the solution by Whatman filter paper. Take 10 ml of the filtrate in a 100 ml volumetric flask. Make the volume up to the mark  with  diluents. Filter this solution with 0.20 m membrane filter before injection.
Condition
Waters Spherissorb ODS1  10 mm 4.6 ´ 250mm./ UV at 278 nm / 8 minutes


Chloramphenicol   / Syrup

Sample
Dilute a weighed quantity containing the equivalent of 0.150 g of Chloramphenicol   to 500 ml with water , mix and allow to stand for 10 minutes . Mix again and immediately dilute 5 ml of the oral suspension to 100 ml with ethanol  (96 %). Measure the absorbance at the maximum at 271 nm . Calculate the content of Chloramphenicol, taking 178 as the value of A (1%, 1 cm) at the maximum at 271 nm .
Calculation
% of content        =   Absorbance  x 100 / conc, in g per 100 ml x 178
      

Chlorhexidine Gluconate / Solution

M. Phase
Take accurately weighted 1.78 g of Sodium Octane   Sulphonate R in a  1000ml volumetric flask add 240 ml of Purified Water then add 105 ml of Glacial Acetic Acid shake for 5min. then add 655 ml Methanol of HPLC Grade. Filter mobile phase with 0.45µm membrane filter and degas for 30 minutes.
Standard 
Take 1.25 ml Chlorhexidine Gluconate standard solution (20% w/v) equivalent to 250 mg of Chlorhexidine Gluconate in
a 50 ml volumetric flask, make volume with  the mobile phase. Take 2 ml of the resulting solution and make 50 ml with
the same solvent. Calculate the concentration in mg/ml. Filter this solution with 0.45 m membrane filter before injection.
Sample 
Take 2 ml sample containing 10 mg of Chlorhexidine Gluconate in a 50ml volumetric flask, make volume with the mobile phase. Calculate the concentration in mg/ml. Filter this solution with 0.45 m membrane filter before injection.
Condition
Waters Spherisorb 10 mm ODS1 4.6 ´ 250mm./ 40º C / UV at 254 nm / 15 minutes.



Chloroquine Phosphate / Tablet

Standard 
Weigh and powder 20 tablets . Dissolve  a quantity of powder equivalent to 250 mg of Chloroquine Phosphate in 10 ml of  M Sodium Hydroxide and extract with four 25 ml quantities of chloroform. Combine the chloroform extracts and evaporate to a volume of about 10 ml. Add 40 ml of anhydrous acetic acid and carry out non aqueous titration, determining the end point potetiometrically. Each ml of 0.1M perchloric acid VS is equivalent to  25.79 mg of Chloroquine Phosphate .
Calculation
% of Chloroquin Phosphate =  No. of ml of 0.1M perchloric acid VS x F x 25.79 X 100 / 250 mg


Chloroquine Phosphate / Syrup

Solvent
Water
Standard 
80 mg ----® 100 ml ----® take 1 ml ----® 50 ml  (Shake 15 minutes)
Sample 
Same as standard. Take absorbance at 343 nm.


Chlorpheniramine Maleate / Syrup

Sample 
Take 10 ml of syrup equivalent to 4 mg of Chlorpheniramine Maleate in a 250 ml separating funnel. Add 10 ml of M sodium hydroxide . Extract two time each with 50 ml quantities of ether. Wash each ether extract with the same 20 ml of water and extract with successive quantities of 20 ,20 and 5 ml of 0.25 M sulphuric acid. Dilute the combined acid extracts to 50 ml with 0.25 M sulphuric acid. Dilute 5 ml to 25 ml with 0.25 M sulphuric acid and measure the absorbance of the resulting solution at the maximum at 265 nm .Calculate the content of chlorpheniramine maleate taking 212 as the value of A (1% , 1 cm ) at the maximum at 265 nm .
% of       Chlorpheniramine maleate  =   Absorbance   x 100 / conc. in gm per 100 ml x 212


Chlorpheniramine Maleate / Tablet

Sample 
Weigh and powder 20 tablets. Dissolve a quantity of powder equivalent to 25 mg of  Chlorpheniramine
Maleate with 50ml volumetric flask. Add 30 ml of 0.05M Sulphuric acid and shake for 15 minutes . Make volume  up  to 50 ml with the same solvent. Mix & filter.Take 8 ml of the filtrate equivalent to 4 mg of chlorpheniramine  Maleate in a 250 ml separating funnel . Add 10ml of 1 M sodium hydroxide . Extract two time each with 50 ml quantities of ether . Wash each ether extract with the same 20 ml of water and extract with successive quantities of 20.20 and 5 ml of 0.25 M sulphuric acid . Dilute the combined acid extracts to 50ml with 0.25 M sulphuric acid. Dilute 5ml to 25 ml with 0.25 M sulphuric acid and measure the absorbance of the resulting solution at the maximum at 265 nm

Calculate content of chlorpheniramine maleate taking 212 as the value of A (1 % , 1 cm ) at the maximum at 265 nm.

% of chlorpheniramine maleate      = Absorbance     x   100 / conc. in gm per 100 ml  x  212


Chlorpromazine Hydrochloride / Tablet & Injection

Standard 
Carry out the following procedure protected from light. To a quantity containing 50 mg of Chlorpromazine Hydrochloride  add sufficient 0.1M hydrochloric acid to produce 100 ml. Dilute 1 ml of the solution to 100 ml with 0.1M hydrochloric acid  and measure the absorbance of the resulting solution at the maximum at 254 nm .
Calculate the content of  Chlorpromazine Hydrochloride  taking 915 as the value of A (1% , 1cm ) at the maximum at 254 nm . 
Calculation
 % of Chlorpromazine HCl  =  Absorbance of Sample ´ 100 / Conc. Of spl. ´ 915                                          


Cisapride  / Syrup

Standard 
Prepare standard in the same concentration as sample, taking cisapride RS.Concentration 0.001 %
Sample 
Take  suspension equivalent to 25 mg of cisapride  in a 250 ml separating funnel. Rinse the pipette with 5 ml of water. Extract two times each with 40 ml of Chloroform. Wash the combined Chloroform extracts with 25 ml of water.  Collect the extracts in a 400 ml beaker and evaporate to dryness on a water bath. Dissolve the residue in 30 ml of Methanol and dilute to 50 ml with methanol. Mix . Dilute 1 ml of the resulting solution to 50 ml with Methanol.Measure the absorbance at 276 nm taking Methanol as reagent blank.


Clindamycin Phosphate

Buffer

Take 13.6 gm of KH2PO4 in a 1000 ml volumetric flask containing 750 ml distilled water and dissolve by stirring. Adjust the pH 4.0 by H3PO4.Finally make the volume 1000 ml by water.
Ratio   
Buffer: Acetonitrile =78: 22                       
Diluent  
Mobile Phase
Standard 
356.07 mg of Clindamycin Phosphate º 300 mg of Clindamycin ----® 50 ml ----® take 5 ml ----® 50 ml (Shake 10 minutes)
Sample 
Same as standard.
Condition
ODS1 10 m 4.6 x 250 mm / UV at 210 nm / 20 minutes


Clobazam / Tablet

Solvent
Methanol
Standard 
20 mg ----® 100 ml ----® take 2 ml ----® 100 ml  (Shake 25 minutes)
Sample 
Same as standard. Take absorbance at   nm.
                                   

                              Clonazepam / Tablet & Solution
                             .
Sample 
Take 1ml  Pase Solution  equivalent to 2.5 mg of Clonazepam   in a 100 ml Amber 
 volumetric flask. And rinse the Pipette with Isopropyl Alcohol.  Add 70 ml of   
Isopropyl Alcohol and shake for 20 minutes. Make the volume up to the mark 
(100 ml) with Isopropyl Alcohol & mix well. (concentration = 0.0025 %).  
Measure the absorbance of sample solution in a UV at a wavelength of 310 
nm using blank solution.
Calculation
 % of Clonazepam  =  Absorbance of Sample ´ 100 / Conc. Of spl. ´ 364                                               


Clopidogrel  + Aspirin / Tablet

Buffer
Dissolve  2.0 gm of  Di-ammonium Hydrogen Orthophosphate &1.0gm of Tetra-n Butyl Ammonium 
Hydrogen Sulfate in 
1000 ml distilled water. Adjust pH 4.0±0.05 with Orthophosphoric acid.
Ratio   
Accetonitrile : Buffer º 60 : 40
Diluents
Accetonitrile : Buffer º 60 : 40
Standard 
48.94 mg  of Clopidogrel Bisulfate º 37.5mg Clopidogrel +37.5 mg Aspirin ----® 50 ml (Shake 10 minutes)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
Sample 
Same as standard.
Condition
C18  5 mm 4.6 ´ 250mm./ UV at 210 nm / 15 minutes.Aspirin 3.20 min.(approx), 
Clopidogrel  8 min.( approx).


Crotamiton + Permithrin / Lotion
For Crotamiton
Standard
Take 100 mg of Crotamiton working standard in a 100 ml volumetric flask. Add 70 ml of cyclohexane. Shake for 15 minutes. Add cyclohexane to make volume 100 ml. Take 1 ml of the solution in a 50 ml volumetric flask. Make the volume up to mark (100 ml) with methanol. Measure the absorbance at 242 nm taking cyclohexane as blank.
Sample

Take 1 ml of the  lotion sample  containing equivalent to 100 mg of Crotamiton in a 100 ml volumetric flask. Add 70 ml of cyclohexane. Shake for 15 minutes. Add cyclohexane to make volume 100 ml. Filter the solution. Take 1 ml of filtered solution in a 50 ml volumetric flask. Make the volume up to mark (50 ml) with cyclohexane. Measure the absorbance at 242 nm taking cyclohexane as blank.

For Permithrin
Ratio   
Methanol : Distilled Water = 80 : 20
Standard
                                    Weigh and transfer about 100 mg working standard of Permethrin in a 100 ml volumetric flask, dissolve in 75 ml of Methanol then shake for 30 minutes.Adjust the volume 100 ml with the same solvent. Filter this solution with 0.2 mm membrane filter before injection.
Sample 
Take 2 ml Lotion sample equivalent to 100 mg of Permethrin in a 100 ml volumetric flask, dissolve in 75 ml of
Methanol and shake for 30 minutes.Filter this solution with Whatman Filter paper and finally filter with 0.2 mm
membrane filter before injection.
Condition
Water Spherisorb , C-18 , 5m 4.6 x 250 mm / 400 C/ UV at 272 nm/15 minutes



1 comment:

  1. Excellent post! I must thank you for this informative read. I hope you will post again soon.Kind regard Calcium Lactate Gluconate#

    ReplyDelete