HPLC ASSAY METHOD FOR CEFUROXIME

Chromatographic System :

Column               : 25cm x 4.6mm, packed with particles of silica(5µm)

Flow rate             : 1.2ml / minute , so that the resolution factors between the peaks  corresponding to the Cefuroxime axetil diastereoisomers A & B  in solution(4)and between the peaks corresponding to Cefuroxime axetil diastereoisomer A & the Cefuroxime axetil A³ -isomer in  solution (2) are each not less than 1.5.

Wavelength                   : 278nm

Temperature       :Ambient

Injection volume : 20µL

Mobile Phase       : A mixture of 38 volumes of Methanol & 62 volumes of

                               0.2M Ammonium dihydrogen orthophosphate.

 Solution (1) :- Disperse 10 tablets in 0.2M Ammonium Dihydrogen Orthophosphate

previously adjusted to pH 2.4 with Orthophosphoric acid using 10ml per gm of the stated content of cefuroxime . Immediately add sufficient methanol to produce a solution containing the equivalent to 0.5%w/v of cefuroxime and shake vigorously.

Filter and dilute a quantity of the filtrate with sufficient mobile phase to produce a

solution containing the equivalent to 0.025% w/v of cefuroxime.

Solution (2) :- Heat a quantity of solution(1) at 60⁰ C for 1hour or until sufficient impurities (   ⁰   -isomers) have been generated.

 Solution (3) :- Expose  a quantity of solution(1) to ultraviolet light (254nm) for            24 hours or until sufficient impurities (E-isomers) have been generated.             Solution (4) contains 0.03% w/v of Cefuroxime Axetil CRS in mobile phase.

  All solutions containing Cefuroxime Axetil, if not to be used for immediate

  analysis, should be stored in the dark at a temperature 2º & 8º before analysis.

The retention times relative to Cefuroxime axetil diastereoisomer A are approximately 0.9 for Cefuroxime axetil diastereoisomer B , 1.2 for the Cefuroxime

axetil  A³-isomers and 1.7 & 2.1 for E-isomers .The relative standard deviation for the

response factor of Cefuroxime axetil for replicate injections of solution (1) is not more than 2.0%.

Calculate the content of C₂₀H₂₂N₄O₁₀S as the sum of the areas of the two peaks  corresponding to diastereoisomers A & B of Cefuroxime axetil using the declared

Content of C₂₀H₂₂N₄O₁₀S in Cefuroxime axetil CRS. Each mg of Cefuroxime axetil

Is equivalent to 0.8313mg Cefuroxime.

Assay of Montelukast Sodium (UV-VIS)Spectrophotometric Method

Assay of Montelukast Sodium (UV-VIS)Spectrophotometric Method.

Preparation of standard solution : Take 50mg of Montelukast Sodium in a 100ml volumetric flask, add 50ml of water and sonicate . Add 10ml of 1M Sodium

Hydroxide solution and dilute to 100ml with water. Transfer 1ml of this solution in a 50ml volumetric flask and dilute to 50ml with o.1N NaOH solution.

Preparation of sample solution :  Crush and finely powder 20 tablets. Weigh an amount of tablet powder containing 50mg of Montelukast Sodium in a 100ml volumetric flask, add 50ml of water and sonicate for about 10 minutes. Add 10ml of 1M NaOH soln. and dilute to 100ml with water, filter. Transfer 1ml of this solution in a 50ml volumetric flask and dilute to 50ml with 0.1N NaOH soln.

Measure the absorbance of the resulting solution at he maximum wavelength 345nm, using 0.1N NaOH as blank.

                                            Abs. of sample X Wt. of standard

% 0f Montelukast Sodium=---------------------------------------X potency of Standard

                                             Abs.of Std. X    Wt. of sample

HPLC ASSAY METHOD OF ASPIRIN

Preparation of Mobile phase :- Dissolve 2gms of sodium 1-heptasulphonate in a mixture of 850ml of water and 150ml of acetonitrile,and adjust with glacial acetic

acid to a pH of 3.4.

Diluting solution : Prepare a mixture of acetonitrile and formic acid (99:1)

Standard preparation : Dissolve an accurately weighed quantity of USP Aspirin RS in diluting solution to obtain a solution having a known concentration of about 0.5mg/ml

Assay preparation:  Weigh and finely powder not less than 20 tablets. Transfer an accurately weighed quantity of powder ,equivalent to about 100mg of aspirin ,to a suitable container. Add 20ml of diluting solution and about 10 beads. Shake vigorously for about 10 minutes, and centrifuse (Stock Solution).

Quantitatively dilute an accurately measured volume of the stock solution with

9 volumes of diluting solution (Assay preparation).

Chromatographic System :

Column                : 4.0mm X 30cm containing packing L1

Flow rate              : 2ml per minute.

Wavelength          : 280nm.     

Temperature                  : Ambient

Injection volume   : about 10µL.

Procedure : Seperately inject an equal volumes (about 10µL) of Standard preparation and Assay preparation into the chromatograph , record the chromatograms,and measure responses for the major peaks.

Calculate the quantity,in mg,of the aspirin (C₉H₈O₄) in the portion of the

tablets taken by the formula :

                                     200C(ru / rs),

In which C is the concentration, in mg per ml, of USP Aspirin RS in the standard preparation , an ru and rs are the peak responses of the aspirin peaks obtained from

the Assay preparation and the Standard preparation respectively.

ASSAY OF CLOPIDOGREL BISULPHATE BY HPLC METHOD

Preparation of phosphate buffer solution : Dissolve 2.72gms of monobasic potassium phosphate in about 500ml of water and dilute to 1000ml with same solvent. Adjust pH to 6.0 ± 0.1 with KOH.

Mobile Phase :  Prepare a filtered and degassed mixture of Phosphate Buffer and acetonitrile (35:65) .Make adjustment if necessary.

Standard Preparation : Dissolve 50mg 0f Clopidogrel Bisulphate Working Standard in methanol, sonicate and make volume to 50ml with methanol.Dilute 5ml of solution to 50ml with mobile phase (conc.0.1mg/ml). Filter the solution with 0.22µm before injection.

Assay Preparation : Crush finely 20 tablets into powder and weigh an amount of powder containg 100mg of Clopidogrel Bisulphate in a 100ml volumetric flask . Dissolve the content with methanol, sonicate and dilute with methanol to volume and mix, filter. Pipette 5ml of this solution to a 50ml volumetric flask, dilute with mobile phase to volume and mix.

Chromatographic System :

Column                  : 4.6mm x 25cm column that contains packing L1

Flow rate              : 1.5ml /min

Wavelength          : 265nm

Temperature                  : Ambient

Injection volume   : 20µL

The relative standard deviation (RSD) for replicate injections determined from Clopidogrel Bisulphate is not more than 1.0% .

Procedure :

Seperately inject equal volumes (about 10µL) of the standard preparation and the assay preparation into the chromatograph ,record the chromatograms, and measure the areas for all peaks . Calculate the quantity in mg of C₁₆H₁₆ClN0₂S.H₂S0₄ in

The portion of the tablet taken by the formula :

                                          Peak area of sample     Wt.of sample

% of Clopidogrel Bisulphate :------------------------X  --------------------X Potency of WS

                                           Peak of standard        Wt.of standard      

Where :  WS is the Working Standard .

Assay of Atorvastatin (UV-VIS) Spectrophotometric Method

Preparation of Standard Solution :  Take 10mg. of Atorvastatin Calcium Trihydrade in a 100ml volumetric flask, add 50ml of Methanol and sonicate , dilute to volume with the same solvent. Transfer 5ml of this solution in a 50ml Volumetric flask and dilute to volume with methanol.

Preparation of Sample Solution : Crush 20 tablets finely and weigh an amount of powder tablet containing 10mg of Atotvastatin Calcium Trihydrade in a 100ml Volumetric Flask ,add 50ml Methanol and sonicate , dilute to Volume with methanol and filter. Transfer 5ml of the filtrate in a 50ml volumetric flask and make volume with methanol.

Measure the absorbance of the resulting solutionat the maximum wavelength at 247nm,using a methanol as blank.

Calculation :

                        

                                Absorbance of sample x weight of standard

% of Atorvastatin = …………………………………………… X Potency of Standard                   

                                 Absorbance of Std.x Weght of sample x 1.084                                                                          

Sampling of Raw Materials

1.0      PURPOSE:  To ensure the smooth sampling of raw materials. It is important that the Sampling is done in an appropriate manner so that a representative sample is taken. 2.0      SCOPE:  Any materials related to manufacturing which is entering the warehouse, Must be sampled properly. 3.0      DEFINITIONS :  Nil 4.0      RESPONSIBILITY: QC officer 5.0      PROCEDURES: 5.1      After receipt of the material in the warehouse the store officer will send an Advice note to the QC department for sampling the material. The advice note Will contain in following information:             5.1.1        Name of the material 5.1.2        Name of the product 5.1.3        Country of origin 5.1.4        Control number of the product 5.1.5        Date of manufacturer 5.1.6        No. of containers received 5.1.7        Description of the containers received 5.1.8        Pack size of the containers.                    Sampling should be done in the following manner:                         5.2.1  Up to three containers        -       from one container                         5.2.2  From 4 to 5 containers       -       from 2 containers                         5.2.3  From 6 to 15 containers     -       from 3 containers                         5.2.4  From 16 to above               -       from every 5th container

Sample should be collected in inert container so that they do not react with the sample. For powdered materials samples should be taken with suitable sampling scoops made up of inert materials. For liquid this should be taken with glass sampling tubes. Suitable siphons may be used in case of sampling thick liquids. Amounts of sample taken should be at least twice the amount required for complete test.