ETHAMBUTOL HYDROCHLORIDE SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the Ethambutol Hydrochloride from
the Pharmacopoeial specifications.
2.0 SCOPE:
This
SOP shall be applicable in Q.C laboratory.
3.0 RESPONSIBILITY:
3.1
Q.C Analysts.
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 (2 test
tubes, 1 spatula, 1 pipette).
5.1.2.1.2
Ethanol (96%).
5.1.2.1.3
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 2 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 ethanol (96%)
in test tube 2 and observe.
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 ethanol (96%) is soluble.
5.2 Identification
tests:
5.2.1
5.2.1.1
Material and equipment:
5.2.1.1.1
Glassware
(according to requirement).
5.2.1.1.2
0.2ml of copper
sulfate solution R.
5.2.1.1.3
10.0ml of Purified
water.
5.2.1.1.4
0.5ml of dilute
sodium hydroxide solution R.
5.2.1.2
Sample:
5.2.1.2.1
0.1g.
5.2.1.3
Method:
5.2.1.3.1
Take a test tube
add in it 0.1g of the sample in it.
5.2.1.3.2
Add 10.0ml of
Purified water R. dissolve it thoroughly.
5.2.1.3.3
Add 0.2ml of
copper sulfate solution R and 0.5ml of dilute sodium hydroxide solution R.
5.2.1.3.4
Observe the
changes.
5.2.1.4
Observations:
5.2.1.4.1
A blue colour is
produced.
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
Centrifugation
machine.
5.2.2.1.3
Dilute nitric
acid.
5.2.2.1.4
Purified water.
5.2.2.1.5
0.4ml of Silver
nitrate R1.
5.2.2.1.6
Ammonia.
5.2.2.2
Sample:
5.2.2.2.1
Quantity of
substance to be examined equivalent to about 2.0mg of chloride.
5.2.2.3
Method:
5.2.2.3.1
Take a test tube
add in it 2.0ml of water with the help of pipette.
5.2.2.3.2
Dissolve in 2.0ml
of water a quantity of the substance to be equivalent to about 2.0mg of
chloride.
5.2.2.3.3
Acidify with
dilute nitric acid.
5.2.2.3.4
And add 0.4ml of
silver nitrate R1.
5.2.2.3.5
Shake and allow it
to stand.
5.2.2.3.6
A curdled, white
ppt is formed.
5.2.2.3.7
Centrifuge it in
centrifugation machine, according to SOP No. BM/QCEO/SOP010-00.
5.2.2.3.8
The obtained ppt
is wash with 3 quantities, each of 1ml, of water.
5.2.2.3.9
Carry out this
operation rapidly is subdued light, degrading the fact that the supernatant
solution may not become perfectly clear.
5.2.2.3.10 Suspend
the precipitate in 2.0ml of water and add 1.5ml of ammonia.
5.2.2.4
Observations:
5.2.2.4.1
The precipitate
dissolves easily with the possible exception of a few large particles which
dissolves slowly.
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
0.5g.
5.3.3
Method:
5.3.3.1
Weigh 0.5g of the
test sample.
5.3.3.2
Set the oven
apparatus. Operate it according to the SOP.
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 at least 3hours.
5.3.3.5
And wait till the
sample loses its moisture.
5.3.3.6
After 3 hours weigh
the sample again by using analytical weighing balance i.e. the final weight.
5.3.3.7
Note down readings
on given Annexure-1.
5.3.4
Observation:
5.3.4.1
Maximum 0.5%.
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
1.0ml of 0.1M
hydrochloric acid.
5.4.2.2
0.1M sodium
hydroxide.
5.4.2.3
Thymolphthalein
solution as an indicator.
5.4.2.4
50.0ml of Purified
water.
5.4.3
Sample:
5.4.3.1
0.200g.
5.4.4
Method
of analysis:
5.4.4.1 Take
a 50.0ml of beaker and 0.200g of sample in it.
5.4.4.2 Add
50.0ml of purified water and add 1.0ml of 0.1M hydrochloric acid in it and dissolve
by using magnetic stirrer operate according to SOP.
5.4.4.3 Fill
the right hand side burette with titrant 0.1M sodium hydroxide.
5.4.4.4 Carry
out a Potentiometric titration using thymolphthalein solution as an indicator.
5.4.4.5 Operate
potentiometer according to SOP.
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-2.
5.4.4.7 Plot
a graph, volume used v/s millivolts.
5.4.4.8 Find
out the END POINT. Read the volume added between the 2 points of inflexion.
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-2.
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 Sodium hydroxide is equivalent to 27.72mg of ethambutol hydrochloride C10H26Cl2N2O2.
5.4.6
Limit:
5.4.6.1
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 / Ethambutol
Hydrochloride: 2015, pp. 895-896.
7.2
The British
Pharmacopoeia. Vol V.,
Official Monograph /Qualitative Reactions and Tests: 2015, pp. 266-270.
8.0 ANNEXURES:
Annexure 1: Observations
of Percentage Loss of drying by using oven.
Annexure 2: Assay
observations and calculations (Potentiometric titration).
Annexure: 1
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: 2
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
|
|
Q.C
|
Quality
control
|
|
%
|
Percentage
|
|
B.P
|
British
pharmacopoeia
|
|
mg
|
Milligram
|
|
ml
|
Milliliter
|
|
ppt
|
Precipitate
|
|
g
|
Grams
|
|
M
|
Molar
|
|
Vol
|
Volume
|
|
QCA
|
Quality
control active ingredient
|
|
F
|
Format
|
|
v/s
|
Verses
|
|
mV
|
Millivolts
|
|
oC
|
Degree
centigrade
|
|
R
|
Reagent
|