1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of
Clopidogrel hydrogen sulphate from the Pharmacopoeial specifications.
2.0 SCOPE:
This
SOP shall be applicable in Q.C laboratory.
3.0 RESPONSIBILITY:
3.1 Q.C Analyst.
4.0 ACCOUNTABILITY:
4.1 Q.C Manager.
5.0 PROCEDURE:
5.1 Characters:
5.1.1
Appearance:
5.1.1.1 White or almost white powder.
5.1.2
Solubility:
5.1.2.1 Material and equipment:
5.1.2.1.1
Glassware
(3 test tubes, 1 spatula, 1 pipette).
5.1.2.1.2
Purified
water.
5.1.2.1.3
Methanol.
5.1.2.1.4
Cyclohexane.
5.1.2.2 Sample:
5.1.2.2.1
Small
quantity.
5.1.2.3 Method:
5.1.2.3.1
Take
3 test tubes and add small quantity of sample for testing solubility according
to B.P specifications.
5.1.2.3.2
Add
purified water, methanol and cyclohexane in each test tube separately in a
small volume and observe the solubility of the sample.
5.1.2.4 Observations:
5.1.2.4.1
The
sample in test tube 1 containing with water is freely soluble.
5.1.2.4.2
The
sample in test tube 2 containing with methanol is freely soluble.
5.1.2.4.3
The
sample in test tube 3 containing with cyclohexane is practically insoluble.
5.2 Identification
tests:
5.2.1
Specific optical rotation:
5.2.1.1 Material and equipment:
5.2.1.1.1
Polarimeter.
5.2.1.1.2
Analytical
weighing balance.
5.2.1.1.3
Glassware
(1 beaker of 50.0ml, 1 stirrer, 1 spatula).
5.2.1.1.4
Methanol.
5.2.1.2 Sample:
5.2.1.2.1
0.250g
of sample.
5.2.1.3 Method:
5.2.1.3.1
Test solution:
5.2.1.3.1.1 Take a beaker of 25.0ml and dissolve
0.250g of sample in sufficient quantity of methanol (approximately
10.0-15.0ml).
5.2.1.3.1.2 Dilute it to 25.0ml with the same solvent.
5.2.1.3.2
Firstly
clean the Polarimeter with clean dry cloth,
5.2.1.3.3
Operate
the Polarimeter0
5.2.1.3.4
Fill
the Polarimeter tube with blank solution and determine the observed optical
rotation.
5.2.1.3.5
Similarly,
fill the Polarimeter tube with sample solution and determine the observed
optical rotation.
5.2.1.3.6
Note
down the values in annexure-2.
5.2.1.3.7
Calculate
the specific optical rotation by using formula:
[α]λ T = α/lc
5.2.1.4 Observations:
5.2.1.4.1
+54.0
to +58.0.
5.2.2
Sulfates test:
5.2.2.1 Material and equipment:
5.2.2.1.1
Glassware
(3 test tubes, 1 pipette).
5.2.2.1.2
Barium
chloride.
5.2.2.1.3
Dilute
hydrochloric acid.
5.2.2.1.4
Purified
water.
5.2.2.2 Sample:
5.2.2.2.1
45.0mg
of sample.
5.2.2.3 Method:
5.2.2.3.1
Test solution:
5.2.2.3.1.1 Take a test tube and add purified water in
small quantity. (approx. 5.0ml)
5.2.2.3.1.2 Dissolve sample in it completely and
perform test on it.
5.2.2.3.2
Add
1.0ml of dilute hydrochloric acid and 1.0ml of barium chloride solution.
5.2.2.3.3
Observe
changes.
5.2.2.4 Observations:
5.2.2.4.1
White
ppt produced.
5.3 Assay:
5.3.1
Apparatus:
5.3.1.1 Glassware (according to requirement).
5.3.1.2 Potentiometer.
5.3.2
Material and reagents:
5.3.2.1 10.0ml of acetone.
5.3.2.2 10.0ml of methanol.
5.3.2.3 30.0ml of purified water.
5.3.2.4 0.1M sodium hydroxide.
5.3.2.5 Thymolphthalein solution as an indicator.
5.3.3
Sample:
5.3.3.1 0.106g of sample.
5.3.4
Method of analysis:
5.3.4.1 Take a 50.0ml of beaker and prepare a
mixture of 10.0ml of acetone, 10.0ml of methanol & 30.0ml of purified
water.
5.3.4.2 Take 0.106g of sample and dissolve it in
above prepare mixture by using stirrer.
5.3.4.3 Fill the right hand side burette with
titrant 0.1M sodium hydroxide
5.3.4.4 Carry out a Potentiometric titration using
Thymolphthalein solution as an indicator.
5.3.4.5 Operate potentiometer.
5.3.4.6 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-1.
5.3.4.7 Plot a graph, volume used v/s millivolts.
5.3.4.8 Find out the END POINT.
5.3.4.9 Peak of graph indicates END POINT i.e. the
point at which maximum millivolts. Note down volume used at that point.
5.3.4.10 Perform blank titration without using
sample. Similarly, as sample titration performed. Record observations in
annexure-1.
5.3.4.11 Calculate volume used by substance by
using formula:
Volume
used by substance = Blank titration - Sample titration.
5.3.4.12 Calculate percentage purity of the sample
by using formula:
%age
purity = volume used by substance x factor x 100
Weight of sample
5.3.5
Factor:
5.3.5.1 1ml of 0.1M Sodium hydroxide is equivalent
to 20.99mg of Clopidogrel
hydrogen sulphate C16H18ClNO6S2.
5.3.6
Limit:
5.3.6.1 99.0% to 101.0% (anhydrous substance).
6.0 REVISION
LOG:
Revision No.
|
Effective Date
|
Reason
|
00
|
New
SOP
|
7.0 REFERENCES:
7.1 The British Pharmacopoeia. Vol I., Official Monograph /Clopidogrel
Hydrogen Sulphate: 2015, pp. 599-601.
8.0 ANNEXURES:
Annexure 1: Assay
observations and calculations (Potentiometric titration).
Annexure 2: Specific
optical rotation observations and calculations.
Annexure:
1
Assay
observations and calculations (Potentiometric titration)
Potentiometric
titration
Reference
electrode: ___________________
Indicator
electrode: ____________________
Speed
of magnetic stirrer: _______________
Titrant
used: __________________________
Indicator:
____________________________
Blank titration:
Plot
a graph, volume used v/s millivolts and find out peak of graph i.e. END POINT
of blank titration.
Sample
titration:
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: ____________________________________________________________
|
Annexure:
2
Specific
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:
Average: _______________
Optical
rotation of blank solution: _______________
Sample solution:
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: ___________________________________________________________
|
9.0
ABBREVIATIONS:
Abbreviation
|
Expanded Form
|
SOP
|
Standard
operating procedure
|
&
|
And
|
No.
|
Number
|
Ltd.
|
Limited
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
Q.C
|
Quality
control
|
mg
|
Milligrams
|
M
|
Molar
|
g
|
Grams
|
ml
|
Milliliter
|
v/s
|
Verses
|
%
|
Percentage
|
Sr.#
|
Serial
number
|
B.P
|
British
pharmacopoeia
|
mV
|
Millivolts
|
Vol
|
Volume
|