MISOPROSTOL SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the
Misoprostol 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
Clear.
5.1.1.2
Colourless or
yellowish.
5.1.1.3
Oily liquid.
5.1.1.4
Hygroscopic.
5.1.2
Solubility:
5.1.2.1
Material and equipment:
5.1.2.1.1
Glassware (test
tubes, spatula).
5.1.2.1.2
Ethanol (96.0%).
5.1.2.1.3
Acetonitrile.
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 ethanol
(96.0%) in test tube 2 and observe.
5.1.2.3.4
Add acetonitrile
in test tube 3 and observe.
5.1.2.4
Observations:
5.1.2.4.1
The sample in test
tube 1 containing with water is practically insoluble.
5.1.2.4.2
The sample in test
tube 2 containing with ethanol (96.0%) is soluble.
5.1.2.4.3
The sample in test
tube 3 containing with acetonitrile is sparingly soluble.
5.2 Assay:
5.2.1
Apparatus:
5.2.1.1
Liquid
chromatography apparatus.
5.2.1.2
Glassware
(according to the requirement).
5.2.1.3
UV/VIS Spectrophotometer
(detector).
5.2.1.4
Sonicator.
5.2.2
Material
and reagents:
5.2.2.1
Silica gel for
chromatography (5μm).
5.2.2.2
Misoprostol CRS.
5.2.2.3
Acetonitrile.
5.2.2.4
Dioxin.
5.2.2.5
Heptane.
5.2.3
Requirements:
5.2.3.1 Sample:
5.2.3.1.1
25.0mg
5.2.3.2 Test solution:
5.2.3.2.1
Test solution:
5.2.3.2.1.1 Take
50.0ml of beaker and dissolve 25.0mg of the substance to be examined in the
mobile phase.
5.2.3.2.1.2 Dilute
it to 5.0ml with the mobile phase.
5.2.3.3 Reference
solutions:
5.2.3.3.1
Reference solution
(a):
5.2.3.3.1.1 Take
100ml beaker and dissolve 25.0mg of Misoprostol CRS in the mobile phase.
5.2.3.3.1.2 Dilute
it to 5.0ml with the mobile phase.
5.2.3.4 Column:
5.2.3.4.1
Size:
5.2.3.4.1.1 Length=0.25m,
5.2.3.4.1.2 θ=4.6mm.
5.2.3.4.2
Stationary
phase:
5.2.3.4.2.1 Silica
gel for chromatography (5μm).
5.2.3.5 Mobile phase:
5.2.3.5.1
Mix 5 volumes of
acetonitrile, 215 volumes of dioxin and 780 volumes of heptane and sonicate for
10min.
5.2.3.6 Flow rate:
5.2.3.6.1
2ml/min.
5.2.3.7 Detection:
5.2.3.7.1
Spectrophotometer
at 210nm.
5.2.3.8 Injection:
5.2.3.8.1
20μL of the test
solution and reference solution (a).
5.2.3.9
Run
time:
5.2.3.9.1
1.5 times the
retention time of the 1st peak of misoprostol.
5.2.3.10
Retention
time:
5.2.3.10.1 1.5
times the retention time of misoprostol.
5.2.3.11
System
suitability
5.2.3.11.1 Reference
solution (a):
5.2.3.11.1.1 Symmetry factor:
maximum 3.7 for the peak due to misoprostol.
5.2.4
Method
of analysis:
5.2.4.1 Firstly
prepare the test solution, reference solution and mobile phase according to the
requirements.
5.2.4.2 The
solutions must be free from solid particles.
5.2.4.3 Prepare
the apparatus.
5.2.4.4 The
mobile phase solvent mixtures must be deaerated prior to use either by boiling
or by applying a partial vacuum to the solvent reservoir.
5.2.4.5 Equilibrate
the column with the prescribed mobile phase, flow rate and at temperature
specified until a suitable baseline is achieved.
5.2.4.6 Test
solution of the mixture to be separated is now introduced into the mobile phase
with the help of an injector just before entering the separating column.
5.2.4.7 As
the eluate leaves the column it enters a detector, where it is continuously
monitored at the specified λ.
5.2.4.8 The
electrical signal obtained from detector is amplified and routes to recorder
which record the developed chromatogram.
5.2.4.9 Calculate
the percentage content of Misoprostol (C22H38O5)
using the declared content of misoprostol CRS.
5.2.5
Limit:
5.2.5.1 96.5%
to 102.0% (anhydrous 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 /Misoprostol: 2015, pp. 298-299.
8.0 ANNEXURES:
Annexure 1: Observations
and calculations of HPLC method.
Annexure:
1
Observations
and calculations of HPLC method
Analysis
on HPLC
Instrument:
___________________ Date: _________________
Model:
___________________
Sample
solution: _______________________
Reference
standard solution: ______________
Impurities:
____________________________
(calculate
each component calculation separately)
OBSERVATIONS:
Attach
chromatogram.
CALCULATIONS:
1.
Retention time: n=
no. of peak
Retention time of unretained peak (tm)=
_____________
2.
Retention volume:
Flow rate= _______________ml/min.
3.
Retention factor:
Retention time of unretained peak (tm)=
_____________
4.
Separation factor (α):
5.
Resolution:
Retention time of unretained peak (tm)=
_____________
6.
Efficiency:
7.
Height equivalent to a theoretical plate (HETP):
Length of column = ________________________
8.
Symmetry factor (tailing factor):
9.
Response factor & Relative response factor:
Conc. (mg/ml)= ___________________
10. Relative standard deviation (%RSD):
Use formula of relative standard deviation where it is
required i.e.,
PIC
11. Percentage of content:
Percentage content = (rU/rS) x (CS/CU)
x 100.
rU= peak response of substance from the sample
solution.
rS= peak response of substance from the standard
solution.
CS= concentration of substance in the standard
solution (mg/mL).
CU= concentration of substance in the sample
solution (mg/mL).
RESULTS:
________________________________________________________________________________________________________________________________________________
|
9.0 ABBREVIATIONS:
Abbreviation
|
Expanded Form
|
SOP
|
Standard
operating procedure
|
&
|
And
|
No.
|
Number
|
Ltd.
|
Limited
|
Sr.#
|
Serial
number
|
Q.C
|
Quality
control
|
%
|
Percentage
|
B.P
|
British
pharmacopoeia
|
UV/VIS
|
Ultraviolet/
visible
|
μm
|
Micron
|
CRS
|
Chemical
reference substance
|
mg
|
Milligram
|
ml
|
Milliliter
|
m
|
Meter
|
θ
|
Theta
|
mm
|
Millimeter
|
λ
|
Lambda
|
Min
|
Minutes
|
ml/min
|
Milliliter
per minutes
|
μL
|
Microliter
|
nm
|
Nanometer
|
Vol
|
Volume
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
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