1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Chloroquine Phosphate 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.
5.1.1.2 Crystalline powder.
5.1.1.3 Hygroscopic.
5.1.2
Solubility:
5.1.2.1 Material and equipment:
5.1.2.1.1
Glassware
(3 test tubes, 1 spatula).
5.1.2.1.2
Alcohol.
5.1.2.1.3
Methanol.
5.1.2.1.4
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
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 in test tube 1 and observe.
5.1.2.3.3
Add
alcohol in test tube 2 and observe.
5.1.2.3.4
Add
methanol in test tube 3 and observe.
5.1.2.4 Observations:
5.1.2.4.1
The
sample in test tube 1 containing with purified water is freely soluble.
5.1.2.4.2
The
sample in test tube 2 & 3 containing with alcohol and methanol is very
slightly soluble respectively.
5.2 Identification
tests:
5.2.1
UV/VIS absorption Spectrophotometry:
5.2.1.1 Material and equipment:
5.2.1.1.1
UV/VIS
Spectrophotometer.
5.2.1.1.2
Glassware
(according to requirement).
5.2.1.1.3
Purified
water.
5.2.1.2 Sample:
5.2.1.2.1
0.100g.
5.2.1.3 Method:
5.2.1.3.1
Test solution:
5.2.1.3.1.1 Take a beaker of 100.0ml and add 0.100g of
sample in it.
5.2.1.3.1.2 Dissolve it in sufficient quantity of purified
water.
5.2.1.3.1.3 And dilute it to 100.0ml with the same solvent.
5.2.1.3.1.4 Take another beaker and add 1.0ml of this
solution in it.
5.2.1.3.1.5 Dilute it to 100.0ml with purified water.
5.2.1.3.2
Spectral range:
5.2.1.3.2.1 Examined between 210nm and 370nm.
5.2.1.3.3
Absorption maxima:
5.2.1.3.3.1 At 220nm, 235nm, 256nm, 329nm and 342nm.
5.2.1.3.4
Operate
the UV/VIS spectrophotometer.
5.2.1.3.5
Examined
between 210nm and 370nm the solution absorption maxima at 220nm, 235nm, 256nm,
329nm and 342nm.
5.2.1.3.6
Note
down values of absorbance in annexure-1.
5.2.1.3.7
Calculate
the specific absorbance by using formula:
1cmA1% = __a__
b.c
5.2.1.4 Observations:
5.2.1.4.1
Specific absorbance:
5.2.1.4.1.1 The specific absorbances at the maxima are
respectively 600 to 660, 350 to 390, 300 to 330, 325 to 355 and 360 to 390.
5.2.2
Melting point determination:
5.2.2.1 Material and equipment:
5.2.2.1.1
Glassware
(according to requirement).
5.2.2.1.2
Melting
point apparatus.
5.2.2.1.3
Capillary
tubes.
5.2.2.1.4
Purified
water.
5.2.2.2 Sample:
5.2.2.2.1
25.0mg.
5.2.2.3 Method:
5.2.2.3.1
Take
a 50.0ml of beaker and add 25.0mg of sample in it.
5.2.2.3.2
Add
20.0ml of purified water and 8.0ml of picric acid solution R1.
5.2.2.3.3
The
ppt, washed with purified water R, with alcohol R and finally with methylene
chloride R.
5.2.2.3.4
Introduce
the sufficient quantity of sample into a capillary tube.
5.2.2.3.5
Set
the apparatus and immerse the capillary tube into the apparatus such that the
closed end is near the center of the bulb of thermometer.
5.2.2.3.6
Switch
on the melting point apparatus.
5.2.2.3.7
Operate
the melting point apparatus.
5.2.2.3.8
Raise
the temperature of the apparatus.
5.2.2.3.9
Record
the temperature at which the last particle passes into the liquid phase.
5.2.2.3.10 Record measurements in annexure-2.
5.2.2.4 Observations:
5.2.2.4.1
The
melting point is 206oC-209oC.
5.2.3
Phosphates determination test:
5.2.3.1 Material and equipment:
5.2.3.1.1
Glassware
(1 test tube, 1 pipette).
5.2.3.1.2
Analytical
weighing balance.
5.2.3.1.3
10.0ml
of purified water.
5.2.3.1.4
2.0ml
of dilute sodium hydroxide solution R.
5.2.3.1.5
Methylene
chloride.
5.2.3.1.6
Nitric
acid.
5.2.3.1.7
2.0ml
of molybdovanadic reagent R.
5.2.3.2 Sample:
5.2.3.2.1
0.1g.
5.2.3.3 Method:
5.2.3.3.1
Take
a test tube and add 0.1g of sample in it.
5.2.3.3.2
Add
10.0ml of purified water R and 2.0ml of dilute sodium hydroxide solution R.
Dissolve it by using magnetic stirrer.
5.2.3.3.3
Shake
with 2 quantities, each of 20.0ml, of methylene chloride R.
5.2.3.3.4
The
aqueous layer is extracted, acidified it by addition of nitric acid R.
5.2.3.3.5
Take
1.0ml of the above prescribed solution and add 2.0ml of molybdovanadic reagent
R
5.2.3.3.6
Observe
the changes.
5.2.3.4 Observations:
5.2.3.4.1
A
yellow colour develops.
5.3 Loss
on drying:
5.3.1
Material
and equipment:
5.3.1.1 Glassware (according to requirement).
5.3.1.2 Analytical weighing balance.
5.3.1.3 Oven.
5.3.2
Sample:
5.3.2.1 1.0g.
5.3.3
Method:
5.3.3.1 Weigh 1.0g of the test sample.
5.3.3.2
Set
the oven apparatus. Operate it.
5.3.3.3 Place the sample into the tray and dry it.
5.3.3.4 Set the temperature of oven at 105oC
for 45min.
5.3.3.5 And wait till the sample loses its
moisture.
5.3.3.6 After 45min weigh the sample again by
using analytical weighing balance i.e. the final weight.
5.3.3.7 Note down readings on given Annexure-3.
5.3.4
Observation:
5.3.4.1 Maximum 2.0%.
5.4 Assay:
5.4.1
Apparatus:
5.4.1.1 Glassware (according to requirement).
5.4.1.2 Potentiometer.
5.4.1.3 Magnetic stirrer.
5.4.2
Material and reagents:
5.4.2.1 50.0ml of anhydrous acetic acid.
5.4.2.2 0.1M Perchloric acid.
5.4.2.3 Crystal violet solution (as indicator).
5.4.3
Sample:
5.4.3.1 0.200g.
5.4.4
Method of analysis:
5.4.4.1 Take a 100.0ml of beaker and take 0.200g
of sample in it.
5.4.4.2 Add 50.0ml of anhydrous acetic acid in it
and dissolve it by using magnetic stirrer.
5.4.4.3 Fill the right hand side burette with
titrant 0.1M Perchloric acid.
5.4.4.4 Carry out a Potentiometric titration using
crystal violet solution as an indicator.
5.4.4.5 Operate potentiometer.
5.4.4.6 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-4.
5.4.4.7 Plot a graph, volume used v/s millivolts.
5.4.4.8 Find out the END POINT.
5.4.4.9 Peak of graph indicates END POINT i.e. the
point at which maximum millivolts. Note down volume used at that point.
5.4.4.10 Perform blank titration without using
sample. Similarly, as sample titration performed. Record observations in
annexure-4.
5.4.4.11 Calculate volume used by substance by
using formula:
Volume
used by substance = Blank titration - Sample titration.
5.4.4.12 Calculate percentage purity of the sample
by using formula:
%age
purity = volume used by substance x factor x 100
Weight of sample
5.4.5
Factor:
5.4.5.1 1ml of 0.1M Perchloric acid is equivalent
to 25.79mg of Chloroquine phosphate C18H32ClN3O8P2.
5.4.6
Limit:
5.4.6.1 98.5% to 101.0% (dried 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 /Chloroquine
Phosphate: 2015, pp. 521.
8.0 ANNEXURES:
Annexure 1:
Observations and Calculations of UV/VIS spectrophotometer.
Annexure 2: Observations
of Melting point apparatus.
Annexure 3: Observations
of Percentage Loss of drying by using oven.
Annexure 4: Assay
observations and calculations (Potentiometric titration).
Annexure: 1
Observations
and Calculations of UV/VIS spectrophotometer
UV/VIS
spectrophotometer
Model:
_____________________________ Date:
_________________
OBSERVATIONS:
CALCULATIONS:
1cmA1% = __a__
b.c
Results:
_______________
Remarks:
______________________________________________________________
|
Annexure: 2
Observations
of Melting point apparatus
Sample
= _____________
Time
period = _____________
Sr.#
|
Initial (Ti)
(oC)
|
Final (Tf)
(oC)
|
Tf - Ti
(oC)
|
Average:
_____________
Result: _________________
Remarks:
_______________________________________________________________
Annexure: 3
Observations
of percentage loss of drying by using Oven
Percentage loss
of drying by using Oven
Weight
of Sample = _____________
Time
period = _____________
Pressure=
_________________
Average
% 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:
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
|
%
|
Percentage
|
B.P
|
British
pharmacopoeia
|
ml
|
Milliliter
|
oC
|
Degree
centigrade
|
mg
|
Milligram
|
g
|
Grams
|
M
|
Molar
|
Min
|
Minutes
|
Vol
|
Volume
|
vi
|
Initial
volume
|
vf
|
Final
volume
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
Ti
|
Initial
temperature
|
Tf
|
Final
temperature
|
Temp.
|
Temperature
|
v/s
|
Verses
|
mV
|
Millivolts
|
UV/VIS
|
Ultraviolet/
visible
|
R
|
Reagent
|
λ
|
Lambda
|