METRONIDAZOLE SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the Metronidazole 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 yellowish.
5.1.1.2
Crystalline
powder.
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
Acetone.
5.1.2.1.3
Alcohol.
5.1.2.1.4
Purified water.
5.1.2.1.5
Methylene
chloride.
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 alcohol in
test tube 3 and observe.
5.1.2.3.5
Add Methylene
chloride in test tube 4 and observe.
5.1.2.4
Observations:
5.1.2.4.1
The sample in test
tube 1, 2, 3 and 4 containing with purified water, acetone, alcohol and
Methylene chloride are slightly soluble respectively.
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 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
is 159oC-163oC.
5.2.2
UV/VIS
absorption Spectrophotometry:
5.2.2.1
Material and equipm4ent:
5.2.2.1.1
UV/VIS
Spectrophotometer.
5.2.2.1.2
Glassware
(according to requirement).
5.2.2.1.3
0.1M hydrochloric
acid
5.2.2.2
Sample:
5.2.2.2.1
40.0mg.
5.2.2.3
Method:
5.2.2.3.1
Test
solution:
5.2.2.3.1.1 Take
a beaker of 100.0ml and add 40.0mg of sample in it.
5.2.2.3.1.2 Dissolve
it in sufficient quantity of 0.1M hydrochloric acid.
5.2.2.3.1.3 And
dilute it to 100.0ml with the same acid.
5.2.2.3.1.4 Take
5.0ml of this solution in 100.0ml of beaker.
5.2.2.3.1.5 And
dilute it to 100.0ml with 0.1M hydrochloric acid.
5.2.2.3.2
Spectral
range:
5.2.2.3.2.1 230-350nm.
5.2.2.3.3
Absorption
maximum:
5.2.2.3.3.1 At
277nm.
5.2.2.3.4
Absorption
minimum:
5.2.2.3.4.1 At
240nm.
5.2.2.3.5
Operate the UV/VIS
spectrophotometer according to the SOP No. BM/QCEO/SOP027-00.
5.2.2.3.6
Measure the
absorption maximum and minimum at given spectral range.
5.2.2.3.7
Measure the
absorbance of the resulting solution at the maximum wavelength 265nm.
5.2.2.3.8
Note down values
of absorbance in annexure-2.
5.2.2.3.9
Calculate the
specific absorbances at the absorption maximum by using formula:
1cmA1% = __a__
b.c
5.2.2.4
Observations:
5.2.2.4.1
Specific
absorbance at absorption maximum at 268nm:
5.2.2.4.1.1 640
to 680nm for the test solution (c).
5.2.3
Primary
aromatic amines test:
5.2.3.1
Material and equipment:
5.2.3.1.1
Glassware (1 test
tube, 1 pipette, 1 spatula).
5.2.3.1.2
Water bath.
5.2.3.1.3
10.0mg of zinc
chloride.
5.2.3.1.4
1.0ml of purified
water.
5.2.3.1.5
0.25ml of dilute
hydrochloric acid.
5.2.3.1.6
0.2ml of sodium
nitrite solution R.
5.2.3.1.7
1.0ml of β-naphthol
solution R.
5.2.3.2
Sample:
5.2.3.2.1
10.0mg.
5.2.3.3
Method:
5.2.3.3.1
Take a test tube
add in it 10.0mg of sample, 10.0mg of zinc chloride R, 1.0ml of purified water
R and 0.25ml of dilute hydrochloric acid.
5.2.3.3.2
Heat it on water
bath for 5 minutes. And after that cool it.
5.2.3.3.3
Acidify the
solution with dilute hydrochloric acid R and add 0.2ml of sodium nitrite
solution R.
5.2.3.3.4
After 1 minute to
2 minute, add 1.0ml of β-naphthol solution R.
5.2.3.3.5
Observe the
changes.
5.2.3.4
Observations:
5.2.3.4.1
An intense orange
or red colour and usually a ppt of the same colour are produced.
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 3 hours.
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-3.
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
50.0ml of anhydrous
acetic acid R.
5.4.2.2
0.1M Perchloric
acid.
5.4.2.3
Crystal violet
solution.
5.4.3
Sample:
5.4.3.1
0.150g.
5.4.4
Method
of analysis:
5.4.4.1 Take
a 50.0ml of beaker and add 0.150g of sample in it.
5.4.4.2 Add
50.0ml of anhydrous acetic acid R in it and dissolve by using magnetic stirrer
SOP No. BM/QCEO/SOP007-00.
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 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-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 17.12mg of Metronidazole C6H9N3O3.
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 II.,
Official Monograph / Metronidazole: 2015, pp. 281-282.
8.0 ANNEXURES:
Annexure 1: Observations
of Melting point apparatus.
Annexure 2: Observations
of UV/VIS spectrophotometer.
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
Sample
= _____________
Time
period = _____________
|
Sr.#
|
Initial (Ti)
(oC)
|
Final (Tf)
(oC)
|
Tf - Ti
(oC)
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Average:
_____________
Result: _________________
Remarks:
_______________________________________________________________
Annexure: 2
Observations
and Calculations of UV/VIS spectrophotometer
|
UV/VIS spectrophotometer
Model:
_____________________________ Date: _________________
OBSERVATIONS:
CALCULATIONS:
1cmA1%
= __a__
b.c
Results:
_______________
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
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
|
|
g
|
Grams
|
|
M
|
Molar
|
|
Vol
|
Volume
|
|
QCA
|
Quality
control active ingredient
|
|
F
|
Format
|
|
v/s
|
Verses
|
|
mV
|
Millivolts
|
|
oC
|
Degree
centigrade
|
|
R
|
Reagent
|
|
Ti
|
Initial
temperature
|
|
Tf
|
Final
temperature
|
|
Temp.
|
Temperature
|
|
λ
|
Lambda
|
|
UV/VIS
|
Ultraviolet/
visible
|
|
nm
|
Nanometer
|
|
h
|
Hours
|
|
A
|
Absorbance
|