To lay down a procedure of analytical report for the active raw material of Ketoconazole from the Pharmacopoeial specifications.
2.0  SCOPE:
This SOP shall be applicable in Q.C laboratory.
3.1  Q.C Analyst.
4.1  Q.C Manager.
5.1  Characters:
5.1.1        Appearance:  White or almost white powder.
5.1.2        Solubility:  Material and equipment:        Glassware (4 test tubes, spatula).        Ethanol (96%).        Methanol.        Methylene chloride.        Purified water.  Sample:        Small quantity.  Method:        Take 4 test tubes and add small quantity of sample for testing solubility according to B.P specifications.        Add purified water in test tube 1, Methylene chloride in test tube 2, methanol in test tube 3, Ethanol (96%) in test tube 4 in a small volume and observe the solubility of the sample.  Observations:        The sample in test tube 1 containing with purified water is practically insoluble.        The sample in test tube 2 containing with Methylene chloride is freely soluble.        The sample in test tube 3 containing with methanol is soluble.        The sample in test tube 4 containing with Ethanol (96%) is sparingly soluble.
5.2  Identification test:
5.2.1        Determination of melting point:  Material and equipment:        Glassware (test tubes, spatula).        Melting point apparatus.        Capillary tubes.  Sample:        Small quantity of sample.  Method:        Introduce the sufficient quantity of sample into a capillary tube.        Set the apparatus and immerse the capillary tube into the apparatus such that the closed end is near the centre of the bulb of thermometer.        Switch on the melting point apparatus.        Operate the melting point apparatus according to the SOP.        Raise the temperature of the apparatus.        Record the temperature at which the last particle passes into the liquid phase.        Record measurements in annexure-1.  Observations:        The melting point of ketoconazole is about 148oC to 152oC.
5.2.2        Chlorides test:  Material and equipment:        Glassware (according to requirement).        Porcelain crucible.        Filter paper.        Bunsen burner.        0.3g of anhydrous sodium carbonate.        Dilute nitric acid.        Purified water (q.s).        0.4ml of Silver nitrate R1.        1.5ml of ammonia.  Sample:        30.0mg.  Method:        Take a porcelain crucible add about 30.0mg of the sample.        Add 0.3g of anhydrous sodium carbonate in it.        Heat it over an open flame for 10 minutes. And allow it cool.        Take up the residue in test tube with 5.0ml of dilute nitric acid.        Filter it by using filtration apparatus.        Take 1.0ml of filtrate in another test tube and add 1.0ml of purified water in it.        Acidify it with dilute nitric acid.        And add 0.4ml of silver nitrate R1.        Shake and allow it to stand.    A curdled, white ppt is formed.    Centrifuge it in centrifugation machine, according to SOP.    The obtained ppt is wash with 3 quantities, each of 1ml, of water.    Carry out this operation rapidly is subdued light, degrading the fact that the supernatant solution may not become perfectly clear.    Suspend the precipitate in 2.0ml of water and add 1.5ml of ammonia.  Observations:        The precipitate dissolves easily with the possible exception of a few large particles which dissolves slowly.
5.3  Solution S:
5.3.1        Material and equipment:  Glassware (1 100.0ml of beaker, 1 spatula, 1 glass rod, 1 pipette).  Analytical weighing balance.  Water bath.  Magnetic stirrer.
5.3.2        Sample:  1.0g.
5.3.3        Preparation of solution S:  Take a beaker of 50.0ml and add sample 1.0g of sample in it.  Add in it sufficient quantity of methylene chloride, dissolve it.  Dilute it to 10.0ml with the same solvent.
5.4  Optical rotation:
5.4.1        Material and equipment:  Polarimeter.  Magnetic stirrer.  Glassware (1 beaker of 50.0ml, 1 stirrer, 1 spatula).  Purified water.
5.4.2        Sample:  Determined on solution S.
5.4.3        Method:  Take a beaker of 50.0ml and add Solution S in it.  Firstly clean the Polarimeter with clean dry cloth, according to SOP No. BM/QCEC/SOP031-00.  Operate the Polarimeter according to SOP.  Fill the Polarimeter tube with blank solution and determine the observed optical rotation.  Similarly, fill the Polarimeter tube with sample solution and determine the observed optical rotation.  Note down the values in annexure-2.  Calculate the specific optical rotation by using formula if needed:
[α]λ T = α/lc  Observations:        Optical rotation of the sample is -0.10o to +0.10o.
5.5  Loss on drying:
5.5.1        Material and equipment:  Glassware (according to requirement).  Analytical weighing balance.  Oven.
5.5.2        Sample:  1.0g.
5.5.3        Method:  Weigh 1.0g of the test sample.  Set the oven apparatus. Operate it according to the SOP.  Place the sample into the tray and dry it.  Set the temperature of oven at 105oC for at least 45 minutes.  And wait till the sample loses its moisture.  After 45 minutes weigh the sample again by using analytical weighing balance i.e. the final weight.  Note down readings on given Annexure-3.
5.5.4        Observation:  Maximum 0.5%.
5.6  Assay:
5.6.1        Apparatus:  Glassware (according to requirement).  Potentiometer.  Magnetic stirrer.
5.6.2        Material and reagents:  Anhydrous acetic acid.  Methyl ethyl ketone.  0.1M Perchloric acid.  Crystal violet solution.
5.6.3        Sample:  0.200g.
5.6.4        Method of analysis:  Take a 100.0ml of beaker and take 0.200g of sample in it.  Add 70.0ml of a mixture of 1 volume anhydrous acetic acid and 7 volumes of methyl ethyl ketone in it and dissolve by using magnetic stirrer.  Fill the right hand side burette with titrant 0.1M Perchloric acid.  Carry out a Potentiometric titration using crystal violet solution as an indicator.  Operate potentiometer according to SOP.  To neutralize analyte add titrant fixed volume (1ml, 0.5ml or 0.1ml) from burette every time note the reading of change in potential difference (millivolts) for each addition in given annexure-4.  Plot a graph, volume used v/s millivolts.  Find out the END POINT.  Peak of graph indicates END POINT i.e. the point at which maximum millivolts. Note down volume used at that point. Perform blank titration without using sample. Similarly, as sample titration performed. Record observations in annexure-4. Calculate volume used by substance by using formula:
          Volume used by substance = Blank titration - Sample titration. Calculate percentage purity of the sample by using formula:
%age purity = volume used by substance x factor x 100
                           Weight of sample
5.6.5        Factor:  1ml of 0.1M Perchloric acid is equivalent to 26.57mg of Ketoconazole C26H28Cl2N4O4.
5.6.6        Limit:  99.0% to 101.0% (dried substance).
Revision No.
Effective Date


7.1  The British Pharmacopoeia. Vol II., Official Monograph /Ketoconazole: 2015, pp. 50-51.
Annexure 1: Observations of Melting point apparatus.
Annexure 2: Optical rotation observations and calculations.
Annexure 3: Observations of Percentage Loss of drying by using oven.
Annexure 4: Assay observations and calculations (Potentiometric titration).

                                                                  Annexure: 1
Observations of Melting point apparatus
Melting point apparatus
Sample = _____________
Time period = _____________
Initial (Ti)
Final (Tf)
Tf - Ti

Average: _____________

Result: _________________

Remarks: ____________________________________________________________ _____________________________________________________________________

Annexure: 2
Optical rotation observations and calculations
Specific optical rotation
Instrument: ___________________                                              Date: _______________
Model: _______________________        Length of Polarimeter tube: ________________
Sample: ________________________________g.
Solvent: ________________________________ml.
Concentration of sample solution: ____________g/ml.
Blank solution:
Blank solution
Optical rotation

                                                                                                 Average: _______________
Optical rotation of blank solution: _______________
Sample solution:
Sample solution
Optical rotation

                                                                                                 Average: _______________
Optical rotation of sample solution: ______________
Optical rotation of substance = Blank solution - Sample solution.

Specific optical rotation of sample solution by using formula:
[α]λ T = α/lc

                                                                      Result: ________________
Remarks: ___________________________________________________________

Annexure: 3
Observations of percentage loss of drying by using Oven
Percentage loss of drying by using Oven
Apparatus: ____________________
Temperature: __________________
Weight of Sample = _____________
Time period = _____________
Pressure= _________________
Time (min)
Weight of sample (g)
% Loss of Moisture
Initial weight
Final weight

Average % Loss of Moisture: _____________

% Loss of Moisture:

Remarks: _______________________________________________________________

Annexure: 4
Assay observations and calculations (Potentiometric titration)
Potentiometric titration
Reference electrode: ___________________
Indicator electrode: ____________________
Speed of magnetic stirrer: _______________
Titrant used: __________________________
Indicator: ____________________________
Blank titration:
Volume used

Plot a graph, volume used v/s millivolts and find out peak of graph i.e. END POINT of blank titration.
Sample titration:
Volume used

Plot a graph, volume used v/s millivolts and find out peak of graph i.e. END POINT of sample titration:
Volume used by Blank titration: __________________
Volume used by Sample titration: _________________
Volume used by substance = Blank titration - Sample titration.

mV used by Blank titration: __________________
mV used by Sample titration: _________________
mV used by substance = Blank titration - Sample titration.

Volume used by substance: _______________________
Voltmeter (mV) used by substance: _________________

RESULT: ____________________________________________________________

Expanded Form
Standard operating procedure
Quality control
British pharmacopoeia
Quality control active ingredient

No comments:

Post a Comment