MONTELUKAST SODIUM SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the
Montelukast sodium 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
White or almost
white.
5.1.1.2
Hygroscopic
powder.
5.1.2
Solubility:
5.1.2.1
Material and equipment:
5.1.2.1.1
Glassware (3 test
tubes, spatula).
5.1.2.1.2
Ethanol (96.0%).
5.1.2.1.3
Methylene
chloride.
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 methylene
chloride in test tube 2 and observe.
5.1.2.3.4
Add ethanol
(96.0%) in test tube 3 and observe.
5.1.2.4
Observations:
5.1.2.4.1
The sample in test
tubes 1 & 2 containing with water and methylene chloride is freely soluble.
5.1.2.4.2
The sample in test
tube 3 containing with ethanol (96.0%) is freely soluble to very soluble.
5.2 Identification
tests:
5.2.1
Sodium
test:
5.2.1.1
Material and equipment:
5.2.1.1.1
Glassware (1
crucible, 1 beaker, 2 test tubes, 1 glass rod, 1 funnel).
5.2.1.1.2
Bunsen burner.
5.2.1.1.3
Filter paper.
5.2.1.1.4
2.0ml of 150g/L
solution of potassium carbonate.
5.2.1.1.5
4.0ml of potassium
pyroantimonate solution.
5.2.1.1.6
Ice water.
5.2.1.1.7
Purified water
(q.s)
5.2.1.2
Sample:
5.2.1.2.1
0.1g.
5.2.1.3
Method:
5.2.1.3.1
Take a crucible
and add 0.1g of sample in it and ignite on Bunsen burner, until an almost white
residue is obtained.
5.2.1.3.2
Take up the
residue in test tube and add 2.0ml of purified water in it.
5.2.1.3.3
Filter it by using
filter paper, funnel and another beaker for collection of filtrate carefully.
5.2.1.3.4
Take 2.0ml of the
above filtrate in test tube.
5.2.1.3.5
2.0ml of the
150g/L solution of potassium carbonate
5.2.1.3.6
Heat it on Bunsen
burner till boiling.
5.2.1.3.7
No ppt is formed.
5.2.1.3.8
Add 4.0ml of
potassium pyroantimonate solution heat on burner till boiling.
5.2.1.3.9
Take a beaker of
ice water. And cool the test tube into it, if necessary rub inside of the test
tube with a glass rod.
5.2.1.3.10 Observe
the changes.
5.2.1.4
Observations:
5.2.1.4.1
A dense white precipitate
is formed.
5.3 Assay:
5.3.1
Apparatus:
5.3.1.1
HPLC apparatus.
5.3.1.2
Glassware
(according to the requirement).
5.3.1.3
Magnetic stirrer.
5.3.2
Material
and reagents:
5.3.2.1
Phenylsilyl silica
gel for chromatography (1.8μm).
5.3.2.2
65.0mg of Montelukast
dicyclohexylamine CRS.
5.3.2.3
Methanol.
5.3.2.4
Trifluroacetic
acid.
5.3.2.5
1000.0ml of acetonitrile.
5.3.2.6
Purified water
(q.s).
5.3.3
Requirements:
5.3.3.1
Sample:
5.3.3.1.1
50.0mg
5.3.3.2
Solvent
mixture: Water and methanol (10:90 v/v).
5.3.3.3
Test
solution:
5.3.3.3.1
Test solution (a):
5.3.3.3.1.1 Take
50.0ml of beaker and dissolve 50.0mg of the substance to be examined in the
solvent mixture.
5.3.3.3.1.2 Dilute
it to 50.0ml with the solvent mixture.
5.3.3.3.2
Test solution (b):
5.3.3.3.2.1 Take
10.0ml of test solution (a) in 100.0ml of beaker.
5.3.3.3.2.2 Dilute
it to 100.0ml with the solvent mixture.
5.3.3.4
Reference
solutions:
5.3.3.4.1
Reference solution
(d):
5.3.3.4.1.1 Take
50.0ml beaker and dissolve 65.0mg of Montelukast dicyclohexylamine CRS in
sufficient solvent mixture. And make up the volume up to 50.0ml with the
solvent mixture.
5.3.3.4.1.2 Take
10.0ml of this solution in another 100.0ml beaker.
5.3.3.4.1.3 Dilute
it to 100.0ml with the solvent mixture.
5.3.3.5
Column:
5.3.3.5.1
Size:
5.3.3.5.1.1 Length=0.05m,
5.3.3.5.1.2 θ=4.6mm.
5.3.3.5.2
Stationary
phase:
5.3.3.5.2.1 Phenylsilyl
silica gel for chromatography (1.8μm).
5.3.3.5.3
Temperature:
5.3.3.5.3.1 30oC.
5.3.3.6
Mobile
phase:
5.3.3.6.1
Mobile
phase A:
5.3.3.6.1.1 Take
1000.0ml of purified water in a beaker of 1000.0ml.
5.3.3.6.1.2 Add
1.5ml of trifluroacetic acid with the help of pipette in beaker.
5.3.3.6.1.3 Mix
it by using magnetic stirrer i.e SOP.
5.3.3.6.2
Mobile
phase B:
5.3.3.6.2.1 Take
1000.0ml of acetonitrile in a beaker of 1000.0ml.
5.3.3.6.2.2 Add
1.5ml of trifluroacetic acid with the help of pipette in beaker.
5.3.3.6.2.3 Mix
it by using magnetic stirrer.
|
Time
(min)
|
Mobile phase A
(percent v/v)
|
Mobile phase B
(percent v/v)
|
|
0-3
|
60
|
40
|
|
3-16
|
60®49
|
40®51
|
5.3.3.7
Flow
rate:
5.3.3.7.1
1.2ml/min.
5.3.3.8
Detection:
5.3.3.8.1
Spectrophotometer
at 238nm.
5.3.3.9
Injection:
5.3.3.9.1
20μL of the test
solution (b) and reference solution (d).
5.3.4
Method
of analysis:
5.3.4.1 Carry
out the test protected from light. Prepare the solutions in amber flasks.
5.3.4.2 Firstly
prepare the test solution, reference solution and mobile phase according to the
requirements.
5.3.4.3 The
solutions must be free from solid particles.
5.3.4.4 Prepare
the apparatus, use normal procedure of liquid chromatography.
5.3.4.5 The
mobile phase solvent mixtures must be deaerated prior to use either by boiling
or by applying a partial vacuum to the solvent reservoir.
5.3.4.6 Equilibrate
the column with the prescribed mobile phase, flow rate and at temperature
specified until a suitable baseline is achieved.
5.3.4.7 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.
5.3.4.8 As
the eluate leaves the column it enters a detector, where it is continuously
monitored at the specified λ.
5.3.4.9 The
electrical signal obtained from detector is amplified and routes to recorder
which record the developed chromatogram.
5.3.4.10 Calculate
the percentage content of Montelukast sodium (C35H3 ClNNaO3S)
using the following expression:
A1 x m2
x 79.24 x p
A2 x m1 x (100-a)
A1
= area of the principal peak in the chromatogram obtained with test
solution (b);
A2
= area of the principal peak in the chromatogram obtained with reference
solution (d);
m1
=mass of the substance to be examined used to prepare test solution (a),
in mg.
m2
=mass of Montelukast dicyclohexylamine CRS used to prepare reference
solution (d), in mg.
p=declared
percentage content of Montelukast dicyclohexylamine CRS.
a=percentage
content of water in the substance to be examined.
5.3.5
Limit:
5.3.5.1 98.0%
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 /Montelukast sodium: 2015, pp. 311-314.
7.2
The British
Pharmacopoeia. Vol V.,
Official Monograph / Liquid Chromatography: 2015, Appendix: IIID pp. 202-204.
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.,
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:
________________________________________________________________________________________________________________________________________________
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9.0 ABBREVIATIONS:
|
Abbreviation
|
Expanded Form
|
|
SOP
|
Standard
operating procedure
|
|
&
|
And
|
|
No.
|
Number
|
|
Ltd.
|
Limited
|
|
Q.C
|
Quality
control
|
|
%
|
Percentage
|
|
g/L
|
Gram
per liter
|
|
ml
|
Milliliter
|
|
q.s
|
Quantity
sufficient
|
|
g
|
Grams
|
|
UV/VIS
|
Ultraviolet/
visible
|
|
ppt
|
Precipitate
|
|
μm
|
Micron
|
|
mg
|
Milligram
|
|
CRS
|
Chemical
reference substance
|
|
v/v
|
Volume
by volume
|
|
m
|
Meters
|
|
θ
|
Theta
|
|
λ
|
Lambda
|
|
oC
|
Degree
centigrade
|
|
Min
|
Minutes
|
|
ml/min
|
Milliliter
per minute
|
|
nm
|
Nanometer
|
|
μL
|
Microliter
|
|
B.P
|
British
pharmacopoeia
|
|
Vol
|
Volume
|
|
QCA
|
Quality
control active ingredient
|
|
F
|
Format
|