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: 45^o.

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 = (r_U/r_S) x (C_S/C_U) x 100

r_U= peak response from the sample solution.

r_S= peak response from the standard solution.

C_S= concentration of the standard solution (mg/ml).

C_U = 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., 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