ISONIAZID SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the Isoniazid 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
Colourless
crystals.
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
Alcohol.
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 alcohol 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 alcohol is sparingly soluble.
5.2 Identification
tests:
5.2.1
Melting
point determination:
5.2.1.1
Material and equipment:
5.2.1.1.1
Glassware
(according to requirement).
5.2.1.1.2
Melting point apparatus.
5.2.1.1.3
Capillary tubes.
5.2.1.1.4
Purified water.
5.2.1.2
Sample:
5.2.1.2.1
Sufficient
quantity of sample.
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.
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
is 170oC-174oC.
5.2.2
5.2.2.1
Material and equipment:
5.2.2.1.1
Glassware (1 test tube,
1 glass rod, 1 pipette).
5.2.2.1.2
Melting point
apparatus.
5.2.2.1.3
2.0ml of purified
water.
5.2.2.1.4
10.0ml of a warm
10g/L solution of vanillin R.
5.2.2.1.5
5.0ml of alcohol
(70% v/v) R.
5.2.2.2
Sample:
5.2.2.2.1
0.1g.
5.2.2.3
Method:
5.2.2.3.1
Take a test tube
add in it 0.1g of the sample in it.
5.2.2.3.2
Add 2.0ml of purified
water in it. Dissolve it thoroughly. And add 10.0ml of a warm 10g/L solution of
vanillin R.
5.2.2.3.3
Allow it to stand
and scratch the wall of the test tube with a glass rod.
5.2.2.3.4
Observe the
changes.
5.2.2.3.5
A yellow ppt is
formed.
5.2.2.3.6
Add 5.0ml of
alcohol (70% v/v) R. Observe changes again.
5.2.2.3.7
It recrystallizes.
5.2.2.3.8
Dry it in oven at
100oC to 105oC.
5.2.2.3.9
Introduce the
sufficient quantity of sample into a capillary tube.
5.2.2.3.10 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.
5.2.2.3.11 Switch
on the melting point apparatus.
5.2.2.3.12 Operate
the melting point apparatus according to the SOP.
5.2.2.3.13 Raise
the temperature of the apparatus.
5.2.2.3.14 Record
the temperature at which the last particle passes into the liquid phase.
5.2.2.3.15 Record
measurements in annexure-2.
5.2.2.4
Observations:
5.2.2.4.1
It melts at 226oC
to 231oC.
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 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 45 minutes.
5.3.3.5
And wait till the
sample loses its moisture.
5.3.3.6
After 45 minutes 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
Not more than 0.1%.
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
20.0ml of hydrochloric
acid.
5.4.2.2
0.2g of potassium
bromide R.
5.4.2.3
0.05ml of methyl
red solution R as an indicator.
5.4.2.4
Purified water.
5.4.2.5
0.0167M Potassium
bromate.
5.4.3
Sample:
5.4.3.1
0.250g.
5.4.4
Method
of analysis:
5.4.4.1 Sample
titration:
5.4.4.1.1
Take a 100.0ml of
beaker and 0.250g of sample in it.
5.4.4.1.2
Add sufficient
quantity of purified water in it and dissolve by using magnetic stirrer operate
according to SOP.
5.4.4.1.3
Dilute it to
100.0ml with the same solvent.
5.4.4.1.4
Take 20.0ml of the
solution in conical flask and add 100.0ml of purified water, 20.0ml of hydrochloric
acid and 0.2g of potassium bromide R.
5.4.4.1.5
Set titration
apparatus.
5.4.4.1.6
Use 0.05ml of
methyl red solution as an indicator.
5.4.4.1.7
Titrate with 0.0167M
Potassium bromate.
5.4.4.1.8
Add dropwise
titrant shake continuously, until the red colour disappears.
5.4.4.1.9
Note down the volume
used as shown in Annexure-4.
5.4.4.1.10 And
take average.
5.4.4.2 Blank
titration:
5.4.4.2.1
Take 20.0ml of the
solution in conical flask and add 100.0ml of purified water, 20.0ml of
hydrochloric acid and 0.2g of potassium bromide R.
5.4.4.2.2
Set titration
apparatus.
5.4.4.2.3
Use 0.05ml of methyl
red solution as an indicator.
5.4.4.2.4
Titrate with 0.0167M
Potassium bromate.
5.4.4.2.5
Add dropwise
titrant shake continuously, until the red colour disappears.
5.4.4.2.6
Note down the
volume used as shown in Annexure-4.
5.4.4.2.7
And take average.
5.4.4.3 Calculate
percentage purity.
5.4.4.4 Calculations:
5.4.4.4.1
After taking
average volume of both blank titration and sample titration. Calculate the
volume used by the examined substance by using formula:
Volume used by
substance = Blank titration - Sample titration.
5.4.4.4.2
For percentage
purity use formula:
%age purity = volume
used by substance x factor x 100
Weight of sample
5.4.4.4.3
Put values and
calculate %age purity.
5.4.5
Factor:
5.4.5.1 1ml
of 0.0167M Potassium bromate is equivalent to 3.429mg of C6H7N3O.
5.4.6
Limit:
5.4.6.1 99.0%
to 101.0%.
6.0 REVISION LOG:
|
Revision No.
|
Effective Date
|
Reason
|
|
00
|
|
New SOP
|
7.0 REFERENCES:
7.1
The British
Pharmacopoeia. Vol I.,
Official Monograph / Isoniazid: 2015, pp. 262.
8.0 ANNEXURES:
Annexure 1: Observations
of Melting point apparatus.
Annexure 2: Observations
of Melting point apparatus.
Annexure 3: Observations
of Percentage Loss of drying by using oven.
Annexure 4: Observations
and calculation of titration.
Annexure: 1
Observations
of Melting point apparatus
Sample
= _____________
Time
period = _____________
|
Sr.#
|
Initial (Ti)
(oC)
|
Final (Tf)
(oC)
|
Tf - Ti
(oC)
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Average:
_____________
Result: _________________
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
Apparatus:
___________________
Temperature:
__________________
Weight
of Sample = _____________
Time
period = _____________
Pressure=
_________________
Average % Loss of Moisture: _____________
Remarks:
_______________________________________________________________
|
||||||||||||||||||||||||
Annexure: 4
Observations
and calculation of titration.
Indicator:
___________________
Weight
of sample: ____________
Factor: ____________
Titrant:
_____________________
Sample titration
|
Sr.#
|
Initial volume (vi)
(ml)
|
Final volume (vf)
(ml)
|
vf-vi
(ml)
|
|
1.
|
|
|
|
|
2.
|
|
|
|
|
3.
|
|
|
|
Average volume: _________________
Blank titration
|
Sr.#
|
Initial volume (vi)
(ml)
|
Final volume (vf)
(ml)
|
vf-vi
(ml)
|
|
1.
|
|
|
|
|
2.
|
|
|
|
|
3.
|
|
|
|
Average volume: _________________
Calculations:
Volume used by substance = Blank
titration - Sample titration.
%age purity = volume used by
substance x factor x 100
Weight of sample
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
|
|
oC
|
Degree
centigrade
|
|
R
|
Reagent
|
|
vi
|
Initial
volume
|
|
vf
|
Final
volume
|