ESCITALOPRAM OXALATE USP SOP
1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Escitalopram Oxalate USP 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 Assay:
5.1.1
Apparatus:
5.1.1.1
Glassware
(according to requirement).
5.1.1.2
Magnetic stirrer.
5.1.1.3
HPLC apparatus.
5.1.1.4
Analytical
weighing balance.
5.1.1.5 Monobasic
potassium phosphate.
5.1.1.6 Phosphoric
acid.
5.1.1.7 Sodium
hydroxide solution.
5.1.1.8 Acetonitrile.
5.1.1.9 USP
Escitalopram oxalate RS.
5.1.1.10 USP
citalopram related compound D RS.
5.1.1.11 Escitalopram
oxalate.
5.1.1.12 Purified
water.
5.1.2
Buffer:
5.1.2.1 Prepare
3.4g/L of monobasic potassium phosphate in purified water.
5.1.2.2 Adjust
with the phosphoric acid or sodium hydroxide solution to a pH of 3.0 before
final dilution.
5.1.3
Solution
A:
5.1.3.1
Prepare solution A
with Acetonitrile and Buffer (10:90) according to the need in a separate
beaker.
5.1.4
Solution
B:
5.1.4.1
Prepare solution B
with Acetonitrile and Buffer (65:35) according to the need in a separate
beaker.
5.1.5
Mobile
phase:
5.1.5.1 See
annexure-1.
[NOTE____
The gradient was established on an HPLC system with a dwell volume of
approximately 1.6ml.]
5.1.6
System
suitability solution:
5.1.6.1 Prepare
2μg/ml each of USP Escitalopram oxalate RS and USP citalopram related compound
D RS in solution A.
5.1.7
Standard
solution:
5.1.7.1 Take
a 25.0ml of volumetric flask and prepare 0.5mg/ml of USP Escitalopram oxalate
RS in solution A.
5.1.8
Sample
solution:
5.1.8.1 Take
a 25.0ml of volumetric flask and prepare 0.5mg/ml of Escitalopram oxalate in
solution A.
5.2 Chromatographic
system:
5.2.1
Mode:
Liquid chromatography.
5.2.2
Detector:
UV
237nm.
5.2.3
Column:
4.6-mm
× 25-cm; 5-μm packing L1.
5.2.4
Column
temperature: 45o.
5.2.5
Flow
rate: See annexure-1.
5.2.6
Injection
size: 20μL.
5.3 System
suitability:
5.3.1
Samples: System
suitability solution and standard solution.
5.3.2
Suitability requirements:
5.3.2.1 Resolution: NLT
1.5 between Escitalopram and citalopram related compound D, system suitability
solution.
5.3.2.2 Tailing factor: 0.8-3, Standard solution.
5.3.2.3 Relative standard
deviation: NMT 2.0%, standard solution.
5.4 Analysis:
5.4.1
Samples: Standard
solution and sample solution.
5.4.2
Calculate the
percentage of Escitalopram oxalate (C20H21FN2O.C2H2O4)
in the portion of Escitalopram oxalate taken:
Result = (rU/rS)
x (CS/CU) x 100
rU=
peak response from the sample solution.
rS=
peak response from the standard solution.
CS=
concentration of the standard solution (mg/ml).
CU
= concentration of the sample solution (mg/ml).
5.5 Procedure:
5.5.1
Equilibrate the
column and detector with mobile phase at specified flow rate until a constant
signal is received.
5.5.2
Separately inject
equal volumes about 20μL of the standard solution and sample solution.
5.5.3
Record the spectrum.
5.5.4
Measure the
responses for the major peaks.
5.5.5
Analyze as
directed in the monograph.
5.6 Limit:
5.6.1
98.0%-102.0% on
the anhydrous basis.
6.0 REVISION LOG:
|
Revision No.
|
Effective Date
|
Reason
|
|
00
|
|
New SOP
|
7.0 REFERENCES:
7.1
USP38NF33
Volume-4 Official Monograph/ Escitalopram Oxalate: 2015, pp.: 3366-3368.
7.2
USP38NF33
Volume-1 Official Monograph/ Chromatography: 2015, pp.: 424-434.
8.0 ANNEXURES:
Annexure 1:
Mobile phase.
Annexure 2: Observations
and calculations of HPLC method.
Annexure:
1
MOBILE
PHASE
|
Time
(min)
|
Solution A
(%)
|
Solution B
(%)
|
Flow rate
(ml/min)
|
|
0
|
95
|
5
|
1
|
|
35
|
65
|
35
|
1
|
|
45
|
0
|
100
|
1
|
|
45.1
|
0
|
100
|
2
|
|
60
|
0
|
100
|
2
|
|
60.1
|
95
|
5
|
1
|
|
68
|
95
|
5
|
1
|
Annexure:
2
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
spectrum.
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:
________________________________________________________________________________________________________________________________________________
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
9.0 ABBREVIATIONS:
|
Abbreviation
|
Expanded Form
|
|
SOP
|
Standard
operating procedure
|
|
&
|
And
|
|
No.
|
Number
|
|
Ltd.
|
Limited
|
|
QCA
|
Quality
control active ingredient
|
|
F
|
Format
|
|
Q.C
|
Quality
control
|
|
Vol
|
Volume
|
|
g
|
Grams
|
|
ml
|
Milliliter
|
|
Min
|
Minutes
|
|
oC
|
Degree
centigrade
|
|
mg
|
Milligram
|
|
M
|
Molar
|
|
USP
|
United
states pharmacopoeia
|
|
NF
|
National
formulary
|
|
cm
|
Centimeter
|
|
nm
|
Nanometer
|
|
mm
|
Millimeter
|
|
Approx.
|
Approximately
|
|
%
|
Percentage
|
|
μL
|
Microliter
|
|
λ
|
Lambda
|
|
mg/ml
|
Milligram
per milliliter
|
|
UV
|
Ultraviolet
|
|
ml/min
|
Milliliter
per minute
|