1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of Betahistine
Dihydrochloride 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 slightly yellow powder, very
hygroscopic.
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
Ethanol.
5.1.2.1.4
2-propanol.
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, ethanol and 2-propanol 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 very soluble.
5.1.2.4.2
The
sample in test tube 2 containing with ethanol is soluble.
5.1.2.4.3
The
sample in test tube 3 containing with 2-propanol is practically insoluble.
5.2 Identification
tests:
5.2.1
Melting point:
5.2.1.1 Material
and equipment:
5.2.1.1.1
Glassware
(test tubes, spatula).
5.2.1.1.2
Melting
point apparatus.
5.2.1.1.3
Capillary
tubes.
5.2.1.2 Sample:
5.2.1.2.1
Small
quantity.
5.2.1.3 Method:
5.2.1.3.1
Introduce
the sufficient quantity of sample into a capillary tube.
5.2.1.3.2
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.
5.2.1.3.3
Switch
on the melting point apparatus.
5.2.1.3.4
Operate
the melting point apparatus according to the SOP No. BM/QCEO/SOP009-00.
5.2.1.3.5
Raise
the temperature of the apparatus.
5.2.1.3.6
Record
the temperature at which the last particle passes into the liquid phase.
5.2.1.3.7
Record
measurements in annexure-1.
5.2.1.4 Observations:
5.2.1.4.1
The
melting point of Betahistine Dihydrochloride is 150oC to 154oC.
5.2.2
Chlorides test:
5.2.2.1 Material
and equipment:
5.2.2.1.1
Glassware
(according to requirement).
5.2.2.1.2
Dilute
nitric acid.
5.2.2.1.3
0.4
ml of Silver nitrate R1.
5.2.2.1.4
Ammonia.
5.2.2.1.5
Purified water.
5.2.2.2 Sample:
5.2.2.2.1
Equivalent
to 2.0mg of chloride
5.2.2.3 Method:
5.2.2.3.1
Take
a test tube add in it 2.0ml of purified water with the help of pipette. And add
a quantity of substance to be examined equivalent to about 2.0mg of chloride.
5.2.2.3.2
Acidify
with dilute nitric acid.
5.2.2.3.3
And
add 0.4ml of silver nitrate R1.
5.2.2.3.4
Shake
and allow it to stand.
5.2.2.3.5
A
curdled, white ppt. is formed.
5.2.2.3.6
Centrifuge
it in centrifugation machine
5.2.2.3.7
The
obtained ppt. is washed with 3 quantities, each of 1ml, of purified water.
5.2.2.3.8
Carry
out this operation rapidly in subdued light, disregarding the fact that the
supernatant solution may not become perfectly clear.
5.2.2.3.9
Suspend
the precipitate in 2.0ml of water and add 1.5ml of ammonia.
5.2.2.3.10 Observe the changes.
5.2.2.4 Observations:
The
precipitate dissolves easily with the possible exception of a few large
particles which dissolves slowly.
5.3 Other
tests:
5.3.1
Solution S:
5.3.1.1 Material
and equipment:
5.3.1.1.1
Glassware
(1 beaker, 1 stirrer, 1 spatula).
5.3.1.1.2
Carbon
dioxide-free water.
5.3.1.2 Sample:
5.3.1.2.1
5.0g.
5.3.1.3 Preparation
of Solution S:
5.3.1.3.1
Take
a beaker of 50.0ml and add 5.0g of the sample in it.
5.3.1.3.2
Add
sufficient quantity of carbon dioxide-free water and dissolve it by using
stirrer.
5.3.1.3.3
Finally
dilute it to 50.0ml with the same solvent.
5.3.2
pH:
5.3.2.1 Material
and equipment:
5.3.2.1.1
Glassware
(according to the requirement).
5.3.2.1.2
pH
meter.
5.3.2.2 Sample:
5.3.2.2.1
Solution
S.
5.3.2.3 Method:
5.3.2.3.1
Firstly
clean the pH meter with clean dry cloth,
5.3.2.3.2
Operate
the pH meter.
5.3.2.3.3
Rejuvenate
the electrode before use according to SOP instructions, if there is any need
of.
5.3.2.3.4
Calibrate
the electrode of the pH meter.
5.3.2.3.5
Perform
the test on solution S.
5.3.2.3.6
Take
a beaker of 100.0ml and add solution S in it. Such that it immersed electrodes
in it completely.
5.3.2.3.7
Maintain
the temperature of sample at 25oC±2oC.
5.3.2.3.8
Dip
the electrode along with temperature sensor into the sample.
5.3.2.3.9
When
dipping electrode into sample, it must be completely immersed in it.
5.3.2.3.10 Stir the probe gently in the sample to
create a homogeneous sample.
5.3.2.3.11 Allow the reading to stabilize for a time.
5.3.2.3.12 Record the observed values of pH &
temperature in the respective Annexure-2.
5.3.2.3.13 Wash the electrodes again after use and
store the electrode in storage solution.
5.3.2.3.14 Calibrate the instrument.
5.3.2.4 Observation:
5.3.2.4.1
2.0
to 3.0.
5.4 Loss
on drying:
5.4.1
Material and equipment:
5.4.1.1 Glassware (according to requirement).
5.4.1.2 Analytical weighing balance.
5.4.1.3 Oven.
5.4.2
Sample:
5.4.2.1 1.0g.
5.4.3
Method:
5.4.3.1 Weigh 1.0g of the test sample.
5.4.3.2 Set the oven apparatus
5.4.3.3 Place the sample into the tray and dry it.
5.4.3.4 Set the temperature of oven at 105oC
for at least 45 minutes.
5.4.3.5 And wait till the sample loses its
moisture.
5.4.3.6 After 45 minutes weigh the sample again by
using analytical weighing balance i.e. the final weight.
5.4.3.7 Note down readings on given Annexure-3.
5.4.4
Observation:
5.4.4.1 Maximum 1.0%.
5.5 Assay:
5.5.1
Apparatus:
5.5.1.1 Glassware (according to requirement).
5.5.1.2 Potentiometer.
5.5.2
Material and reagents:
5.5.2.1 50.0ml of ethanol.
5.5.2.2 0.1M sodium hydroxide.
5.5.2.3 Thymolphthalein solution as an indicator.
5.5.3
Sample:
5.5.3.1 80.0mg of sample.
5.5.4
Method of analysis:
5.5.4.1 Take a titration flask and add 80.0mg of
sample in it.
5.5.4.2 Add 50.0ml of ethanol (96%) R and dissolve
it by using magnetic stirrer
5.5.4.3 Fill the right hand side burette with
titrant 0.1M sodium hydroxide
5.5.4.4 Carry out a Potentiometric titration using
Thymolphthalein solution as an indicator.
5.5.4.5 Operate potentiometer.
5.5.4.6 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.
5.5.4.7 Plot a graph, volume used v/s millivolts.
5.5.4.8 Find out the END POINT.
5.5.4.9 Peak of graph indicates END POINT i.e. the
point at which maximum millivolts. Note down volume used at that point.
5.5.4.10 Perform blank titration without using
sample. Similarly, as sample titration performed. Record observations in
annexure-4.
5.5.4.11 Calculate volume used by substance by
using formula:
Volume
used by substance = Blank titration - Sample titration.
5.5.4.12 Calculate percentage purity of the sample
by using formula:
%age
purity = volume used by substance x factor x 100
Weight of sample
5.5.5
Factor:
5.5.5.1 1ml of 0.1M Sodium hydroxide is equivalent
to 10.46mg of Betahistine
Dihydrochloride C8H14Cl2N2.
5.5.6
Limit:
5.5.6.1 99.0% 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 /Betahistine
Dihydrochloride: 2015, pp. 275-276.
8.0 ANNEXURES:
Annexure 1: Observations
of Melting point apparatus.
Annexure 2: pH
measurement.
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 = _____________
Average:
_____________
Result:
_________________
Remarks:
____________________________________________________________
_____________________________________________________________________
|
Annexure:
2
pH
measurement
Sr.#
|
Temperature
|
pH
|
Result: _________________________________________________________________
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
|
oC
| Centigrade |
QCA
|
Quality
control active ingredient
|
QCEO
|
Quality
Control Equipment (Operation)
|
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
|
oC
|
Centigrade
|