PHOLCODINE METHOD OF ANALYSIS SOP
1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Pholcodine 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.
5.1.1.2
Crystalline
powder.
5.1.1.3
Colourless.
5.1.2
Solubility:
5.1.2.1
Material and equipment:
5.1.2.1.1
Glassware (4 test
tubes, 1 spatula, 1 pipette).
5.1.2.1.2
Ethanol (96%).
5.1.2.1.3
Acetone.
5.1.2.1.4
Dilute mineral acid
(e.g. H2SO4).
5.1.2.1.5
Purified water.
5.1.2.2
Sample:
5.1.2.2.1
Small quantity.
5.1.2.3
Method:
5.1.2.3.1
Take 4 test tubes
and add small quantity of sample for testing solubility according to B.P
specifications.
5.1.2.3.2
Add purified water
in test tube 1 and observe.
5.1.2.3.3
Add acetone in
test tube 2 and observe.
5.1.2.3.4
Add ethanol (96%)
in test tube 3 and observe.
5.1.2.3.5
Add dilute mineral
acid in test tube 4 and observe.
5.1.2.4
Observations:
5.1.2.4.1
The sample in test
tube 1 containing with purified water is sparingly soluble.
5.1.2.4.2
The sample in test
tube 2 and 3 containing with acetone and ethanol (96%) is freely soluble
respectively.
5.1.2.4.3
The sample in test
tube 4 containing with dilute mineral acid is dissolved.
5.2 Other
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
Magnetic stirrer.
5.2.1.1.3
Glassware (1
beaker of 50.0ml, 1 stirrer, 1 spatula).
5.2.1.1.4
Ethanol (96%).
5.2.1.2
Sample:
5.2.1.2.1
1.0g.
5.2.1.3
Method:
5.2.1.3.1
Take a beaker of
50.0ml and add 1.0g of sample in it.
5.2.1.3.2
Dissolve it in
sufficient quantity of ethanol (96%) by using magnetic stirrer.
5.2.1.3.3
And finally dilute
it to 50.0ml with the same solvent.
5.2.1.3.4
Firstly clean the
Polarimeter with clean dry cloth, according to SOP
5.2.1.3.5
Operate the
Polarimeter according to SOP
5.2.1.3.6
Fill the
Polarimeter tube with blank solution and determine the observed optical
rotation.
5.2.1.3.7
Similarly, fill
the Polarimeter tube with sample solution and determine the observed optical
rotation.
5.2.1.3.8
Note down the
values in annexure-1.
5.2.1.3.9
Calculate the
specific optical rotation by using formula:
[α]λ T = α/lc
5.2.1.4
Observations:
5.2.1.4.1
-98 to -94 (dried substance).
5.2.2
Loss
on drying:
5.2.2.1
Material and equipment:
5.2.2.1.1
Glassware
(according to requirement).
5.2.2.1.2
Analytical
weighing balance.
5.2.2.1.3
Oven.
5.2.2.2
Sample:
5.2.2.2.1
0.5g.
5.2.2.3
Method:
5.2.2.3.1
Weigh 1.0g of the
test sample.
5.2.2.3.2
Set the oven
apparatus. Operate it according to the SOP.
5.2.2.3.3
Place the sample
into the tray and dry it.
5.2.2.3.4
Set the
temperature of oven at 105oC for at least 45 minutes.
5.2.2.3.5
And wait till the
sample loses its moisture.
5.2.2.3.6
After 45 minutes
weigh the sample again by using analytical weighing balance i.e. the final
weight.
5.2.2.3.7
Note down readings
on given Annexure-2.
5.2.2.4
Observation:
5.2.2.4.1
3.9% to 4.5%.
5.3 Assay:
5.3.1
Apparatus:
5.3.1.1
Glassware
(according to requirement).
5.3.1.2
Potentiometer.
5.3.1.3
Magnetic stirrer.
5.3.2
Material
and reagents:
5.3.2.1
50.0ml of
anhydrous acetic acid.
5.3.2.2
0.1M Perchloric
acid.
5.3.2.3
Crystal violet solution.
5.3.3
Sample:
5.3.3.1
0.180g.
5.3.4
Method
of analysis:
5.3.4.1 Take
a 50.0ml of beaker and take 0.180g of sample in it.
5.3.4.2 Add
50.0ml of anhydrous acetic acid in it and dissolve by using magnetic stirrer
i.e. SOP.
5.3.4.3 Fill
the right hand side burette with titrant 0.1M Perchloric acid.
5.3.4.4 Carry
out a Potentiometric titration using crystal violet solution as an indicator.
5.3.4.5 Operate
potentiometer according to SOP.
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-3.
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-3.
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 Perchloric acid is equivalent to 19.93mg of Pholcodine C23H30N2O4.
5.3.6
Limit:
5.3.6.1
98.5% to 101.5%
(dried substance).
6.0 REVISION LOG:
Revision No.
|
Effective Date
|
Reason
|
00
|
New SOP
|
7.0 REFERENCES:
7.1 The British Pharmacopoeia. Vol II., Official Monograph / Pholcodine: 2015, pp. 570-571.
8.0 ANNEXURES:
Annexure 1: Specific
optical rotation observations and calculations.
Annexure 2: Observations
of Percentage Loss of drying by using oven.
Annexure 3: Assay
observations and calculations (Potentiometric titration).
Annexure: 1
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:
___________________________________________________________
|
Annexure:
2
Observations
of percentage loss of drying by using Oven
Percentage
loss of drying by using Oven
Apparatus:
____________________
Temperature:
__________________
Weight
of Sample = _____________
Time
period = _____________
Pressure=
_________________
Average % Loss of Moisture: _____________
Remarks:
_______________________________________________________________
|
||||||||||||||||||||||||
Annexure:
3
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: ____________________________________________________________
|
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
|
Vol
|
Volume
|
Ti
|
Initial
temperature
|
Tf
|
Final
temperature
|
Temp.
|
Temperature
|
v/s
|
Verses
|
mV
|
Millivolts
|
g
|
Grams
|
ml
|
Milliliter
|
Min
|
Minutes
|
oC
|
Degree
centigrade
|
mg
|
Milligram
|
M
|
Molar
|
%
|
Percentage
|
R
|
Reagent
|
o
|
Degree
(angle)
|
l
|
Length
|
c
|
Concentration
(g/ml)
|
g/ml
|
Gram
per milliliter
|
α
|
Alpha
|
λ
|
Lambda
|