ESCITALOPRAM OXALATE USP SOP


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: ___________________           
Column size:
Length=
θ=
Stationary phase:

Temperature:

Mobile phase:

Flow rate:

Injection size:

Detector:

Wavelength:
λ=

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)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Height of peak of interest
(h)n
Width of peak of interest
(w)n
Area of peak of interest
A=1/2(h x w)




















2.      Retention volume:
Flow rate= _______________ml/min.
No. of peaks
Retention time of peak of interest
(tr)n
Retention volume = retention time x flow rate












3.      Retention factor:
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Retention factor of a component
k= (tr-tm)/tm













4.      Separation factor (α):
No. of peaks
Retention factor of a component
(kn)
Relative retention of two adjacent peaks
α = k2/k1












5.      Resolution:
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Width of peak of interest
(w)n
Resolution
Rs = 2 (tr2-tr1)
        (w1-w2)
















6.      Efficiency:
No. of peaks or components
Retention time of peak of interest
(tr)n
Width of peak of interest
(w)n
Efficiency
(No. of theoretical plates)
N= 16 (tr/w)2


















7.      Height equivalent to a theoretical plate (HETP):
Length of column = ________________________
No. of peaks or components
No. of theoretical plates
(N)
Height equivalent to a theoretical plate HETP = L/N












8.      Symmetry factor (tailing factor):
No. of peaks or components
Distance from the peak max. to leading edge of the peak
(f)
Width w
Symmetry factor
At 5%
At 10%
As = w5%
       2f
As = w10%
       2f
























9.      Response factor & Relative response factor:
Conc. (mg/ml)= ___________________
No. of peak
Peak area
Response factor = (peak area/conc.)
Relative response factor = (response factor of impurity/response factor of API)

















10.  Relative standard deviation (%RSD):
Use formula of relative standard deviation where it is required i.e.,
https://encrypted-tbn0.gstatic.com/images?q=tbn:ANd9GcSaQVcHe6gxvev49TCGtnMo64gnQ-6-Ilt9DWxADmbQyzxMgA8UFg
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


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