1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the
Crotamiton 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 Colourless or pale yellow.
5.1.1.2 Oily liquid.
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%).
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.
5.1.2.3.4
At
low temperature it may partly or completely soluble.
5.1.2.4 Observations:
5.1.2.4.1
The
sample in test tube 1 containing with water is slightly soluble.
5.1.2.4.2
The
sample in test tubes 2 containing with ethanol (96%) is miscible.
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
UV/Vis
spectrophotometer.
5.2.1.2 Sample:
5.2.1.2.1
25.0mg.
5.2.1.3 Test solution:
5.2.1.3.1
Take
a beaker of 100.0ml. Take sample 25.0mg dissolve it in cyclohexane small volume
of liquid.
5.2.1.3.2
Dilute
it to 100.0ml with the same solvent.
5.2.1.3.3
Again
dilute 1.0ml of this solution to 10.0ml with cyclohexane.
5.2.1.4 Spectral range:
5.2.1.4.1
220-300nm.
5.2.1.5 Absorption maximum:
5.2.1.5.1
At
242nm.
5.2.1.6 Specific absorbance at the maximum absorption:
5.2.1.6.1
300-330.
5.2.1.7 Method of analysis:
5.2.1.7.1
According
to SOP of UV/visible spectrophotometer operate i.e. and observe the absorbance
of test solution.
5.2.1.8 Observations:
5.2.1.8.1
Maximum
absorption found at 242nm.
5.2.2
5.2.2.1 Material and equipment:
5.2.2.1.1
Glassware
(test tube, spatula).
5.2.2.1.2
3g/L
solution of potassium permanganate.
5.2.2.1.3
Purified
water.
5.2.2.2 Sample:
5.2.2.2.1
Saturated
solution of active ingredient.
5.2.2.3 Method of analysis:
5.2.2.3.1
Take
a test tube and add in it 10ml of a saturated solution.
5.2.2.3.2
Add
a few drops of 3g/L solution of potassium permanganate.
5.2.2.3.3
Observe
the changes.
5.2.2.4 Observations:
5.2.2.4.1
A
brown colour is produced.
5.2.2.4.2
A
brown ppt is formed on standing.
5.3 Assay:
5.3.1
Apparatus:
5.3.1.1 HPLC apparatus.
5.3.1.2 Glassware (according to the requirement).
5.3.2
Material and reagents:
5.3.2.1 Tetrahydrofuran 8 volume.
5.3.2.2 Cyclohexane 92 volume.
5.3.3
Requirements:
5.3.3.1 Sample:
5.3.3.1.1
50.0mg
(sample to be examined).
5.3.3.1.2
50.0mg
(Crotamiton CRS) for reference solution.
5.3.3.2 Test
solution:
5.3.3.2.1
Test
solution (a):
5.3.3.2.1.1 Take 100ml of beaker and dissolve 50.0mg
of the substance to be examined in the mobile phase.
5.3.3.2.1.2 And dilute to 100.0ml with the mobile
phase.
5.3.3.2.2
Test
solution (b):
5.3.3.2.2.1 Take a beaker and dilute 1.0ml of test
solution (a) to 20.0ml with the mobile phase.
5.3.3.3 Reference
solutions:
5.3.3.3.1
Reference
solution (a):
5.3.3.3.1.1 Take 100ml beaker and dissolve 50.0mg of Crotamiton
CRS in the mobile phase.
5.3.3.3.1.2 And dilute to 20.0ml with the mobile
phase.
5.3.3.3.1.3 Dilute 1.0ml of this solution to 50.0ml
with the mobile phase.
5.3.3.4 Column:
5.3.3.4.1
Size:
5.3.3.4.1.1 Length=0.25m,
5.3.3.4.1.2 θ=4mm.
5.3.3.4.2
Stationary phase:
5.3.3.4.2.1 Silica gel for chromatography (5μm).
5.3.3.5 Mobile
phase:
5.3.3.5.1
Tetrahydrofuran,
cyclohexane (8:92 v/v)
5.3.3.6 Flow
rate:
5.3.3.6.1
1.0ml/min.
5.3.3.7 Detection:
5.3.3.7.1
Spectrophotometer
at 242nm.
5.3.3.8 Injection:
5.3.3.8.1
20μL
of the test solution (b) and reference solution (a).
5.3.3.9 Run
time:
5.3.3.9.1
2.5
times the retention time of the (E)-isomer.
5.3.3.10 Relative
retention:
5.3.3.10.1 With reference to (E)-isomer:
(Z)-isomer=about 0.5.
5.3.4
Method of analysis:
5.3.4.1 Firstly prepare the test solution,
reference solution and mobile phase according to the requirements.
5.3.4.2 The solutions must be free from solid
particles.
5.3.4.3 Prepare the apparatus.
5.3.4.4 The mobile phase solvent mixtures must be
de-aerated prior to use either by boiling or by applying a partial vacuum to
the solvent reservoir.
5.3.4.5 Equilibrate the column with the prescribed
mobile phase, flow rate and at temperature specified until a suitable baseline
is achieved.
5.3.4.6 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.
5.3.4.7 As the eluate leaves the column it enters
a detector, where it is continuously monitored at the specified λ.
5.3.4.8 The electrical signal obtained from
detector is amplified and routes to recorder which record the developed
spectrum.
5.3.4.9 Calculate the percentage content of Crotamiton
(C13H17NO) from the sum of the areas of peaks due to (Z)
- and (E)-isomers in the spectrum obtained.
5.3.4.10 Calculate the content of the (Z)-isomer,
as a percentage of the total content of the (E) - and (Z)-isomers, from the
chromatogram obtained with test solution (b).
5.3.5
Observations:
5.3.5.1 Sum
of the (E)- and (Z)-isomers:
5.3.5.1.1
96%
to 102%.
5.3.5.2 (Z)-isomer:
5.3.5.2.1
15%.
6.0 REVISION
LOG:
Revision No.
|
Effective Date
|
Reason
|
00
|
New
SOP
|
7.0 REFERENCES:
7.1 The British Pharmacopoeia. Vol I., Official Monograph /Crotamiton: 2015, pp. 655-657.
8.0 ANNEXURES:
8.1 Not Applicable.
9.0 ABBREVIATIONS:
Abbreviation
|
Expanded Form
|
SOP
|
Standard
operating procedure
|
&
|
And
|
No.
|
Number
|
Ltd.
|
Limited
|
Q.C
|
Quality
control
|
%
|
Percentage
|
B.P
|
British
pharmacopoeia
|
UV
|
Ultraviolet
|
mg
|
Milligram
|
ml
|
Milliliter
|
nm
|
Nanometer
|
g/L
|
Gram
per liter
|
ppt
|
Precipitate
|
CRS
|
Control
reference solution
|
g
|
Grams
|
θ
|
Theta
|
μm
|
Micrometer
|
m
|
Meter
|
v/v
|
Volume
by volume
|
ml/min
|
Milliliter
per minute
|
μL
|
Microliter
|
%
|
Percentage
|
λ
|
lamda
|
Vol
|
Volume
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
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