Watson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL | Validation Protocol No. Product: | SIMVASTATIN USP VPKAD100076 | Version No:00 . | __WAT/ARDIRMS/0099(T)-00 Effective date: rl Method : RESIDUAL SOLVENTS BY GAS 16 JUL 20 CHROMATOGRAPHY Page No.: 1 of 23, 1.0 APPROVAL |_| Prepared By-User Department | | _ap® | Anil Me kumbhay Se | Name Signature / Date Reviewed By-Head of the user Department (Designee, Roreah ~S. Rely] BP ea cote Name Signature / Date yo Meyus OR resarelran gia Name Signature / Date | Approved By-Head Quality Assurance / Designee | | CQA025/F01-01Watson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL. MULED COPY ms Validation Protocol No.: Product : ‘SIMVASTATIN USP VPKAD100076 Version No:00 . WAT/ARD/RMS/0099(T)-00 Effective dat 201 | Method : RESIDUAL SOLVENTS BY GAS 6 JUL CHROMATOGRAPHY Page No.: 2 of 23 TABLE OF CONTENTS Contents Approval | 2.0 _ | Abbreviations | 3 Introduction 3 | | Objective 4 5.0 _| Background 4 | 6.0 | Method of Analysis 5 | | Experimental Outline 10 | Specificity | 10 | Quantitation Limit (LOQ) [8 Linearity and Range 15 Accuracy 417 | | 7.5 | Precision | 19 [_75] Robustness _ td CQA025/F02-01 | Approved By (Signature): __ werWatson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL | Product : Validation Protocol No.: ‘SIMVASTATIN USP VPKAD100076 { | Version No:00 Method : WATIARD/RMS/0099(T)-00 Effective date: a 6 yur | RESIDUAL SOLVENTS BY GAS. }—————__——— | CHROMATOGRAPHY Page No.: 3 of 23 2.0 ABBREVIATIONS Full Desi cription ‘Standard Operating Procedure Quantitation Limit Signal to Noise Relative Standard Deviation ~ | willliter Microgram Parts per milion Degree Celsius Gas Chromatography Micrometers. ‘Minutes ‘Grams Number Weight __ [Global Stan: ison | With respect to Tetrahydrofuran THE 3.0 INTRODUCTION ‘Simvastatin is a hypolipidemic drug. It is used to control hypercholesterolemia and to prevent cardiovascular disease. Simvastatin is butanoic acid, 2,2-dimethyl-,1,2,3,7,8,8a-hexahydro-3,7-dimethyl-8-[2- (tetrahy-dro-4-hydroxy-6-ox0-2H-pyran-2-yl)-ethyll [1a,30,78,88(2S* 48"),-Bat its molecular weight is 418.57. CQA025/F02-01 Approved By (: Concentration - J]. The molecular formula of simvastatin is C25H3805 and [, (Signature) ke _ |-naphthalenyl ester, [1S-Watson _India Ambernath (Pharma) | ANALYTICAL METHOD VALIDATION PROTOCOL T Validation Protocol No.: Product : SIMVASTATIN USP VPKAD100076 - Version No:00 | __WATIARDIRMS/0099(T)-00 Effective da 10 Method : RESIDUAL SOLVENTS BY GAS ul 20 CHROMATOGRAPHY Page No.: 4 of 23, ‘The structural formula of Simvastatin is: HO, oO Simvastatin is a white to off-white, nonhygroscopic, crystalline powder that is practically insoluble in water, and freely soluble in chloroform, methanol and ethanol. AGC method has been developed for the determination of residual solvents in Simvastatin. This protocol is intended to demonstrate the suitability of this method for its routine use. | The validation methodology and the acceptance criteria mentioned in this protocol are | in accordance with the GSOP no, GSOP 022-001 Revision : 00, “Method Validation’ (effective date 25 January 2010) 4.0 OBJECTIVE To demonstrate that the proposed GC method, according to the test procedure is suitable for determination of residual solvents in Simvastatin. 5.0 BACKGROUND j This test procedure to be validated is developed at Watson Pharma Pvt. Lid. (Unit Il), Ambernath, CQA025/F02-01 (approved By Sipnatrey __ — Approved By (Signature): gerWatson _India Ambernath (Pharma) __ ANALYTICAL METHOD VALIDATION PROTOCOL Product : Validation Protocol No.: SIMVASTATIN USP VPKAD100076 __ Version No:00 Method : WATIARD/RMS/0099(T)-00 Effective date: 4 6 JUL a RESIDUAL SOLVENTS BY GAS CHROMATOGRAPHY Page No.: 5 of 23 Proposed Specification Level for Methanol, Petroleum ether, Ethyl acetate, ‘Tetrahydrofuran and Toluene in Simvastatin. Methanol NMT 200 ppm Petroleum Ether: NMT 50 ppm Ethyl acetate NMT 2500 ppm. Tetrahydrofuran —: NMT 200 ppm Toluene NMT 200 ppm 6.0 METHOD OF ANALYSIS 6.1. Instruments/Equipment 6.1.4 A gas chromatograph equipped with attachment for capillary column, a flame ionization detector, a head space analyzer and data management system. (Agilent 6890N gas chromatograph with EZChrome Elite software and Combipal H.S. auto sampler is suitable.) | 6.1.2 Semi micro analytical balance (Sartorius, Model ME235 or equivalent). 6.2 Reagents and Chemicals 6.2.1. Methanol GC grade 6.2.2. Petroleum Ether -GR grade 6.2.3. Ethyl acetate -GC grade 6.2.4. Tetrahydrofuran ~ GC grade 62.5. Toluene - GC grade 62.6. _N,N-Dimethyiformamide ~ GC grade 6.3 Chromatographic System 6.3.1. Column DB-624 (60m X 0.53mm X 3ym), (Make — Agilent Technologies) 6.3.2. Carrier gas Nitrogen 6.3.3. Column mode Constant flow 6.3.4. Flow rate 6.5mU min 6.3.5. Column Oven Temperature | Set the temperature program as follows Initial temperature 40°C Hold time 40 min. Approved By (Signature): Se CQA025/F02-01Watson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL —T Validation Protocol No.: Product : SIMVASTATIN USP VPKAD100076 = = Version No:00 0 | . WATI/ARD/RMS/0099(T)-00 Effective date; [16 JUL Method : RESIDUAL SOLVENTS BY GAS ive dato: 6 JUL CHROMATOGRAPHY Page No.: 6 of 23 - a | Ramprate 2 40°C per min. Temperature : 150°C. Hold time 5 min, Ramp rate 2 40°C per min. Final temperature 1 240°C Hold time 3 min, 6.3.6. _Injector temperature > 480°C 6.3.7. Detector temperature : 260°C 63.8. Spiitratio 34 6.3.9. Hydrogen flow 30 mUmin 6.3.10, Air flow 300 mL/min 6.3.11. Make up flow 2 45mUimin 6.3.12, Run time 34.25 min 6.4 Headspace conditions 6.4.1 Incubation temperature : 90°C 6.4.2 Incubation time > 15min 6.4.3 Syringe temperature > 120°C 6.4.4 Agitator speed 500 rpm 645 Fillspeed (uL/s) 400 64.6 Fill stroke 0 64.7 Pullup delay 4000 ms | 64.8 Injection speed 2 800 pLls 649 Pre inject delay : 600ms 6.4.10 Post inject delay : Oms 6.4.11 Injection volume : 1.5 mL of the headspace 6.5 Preparation of solutions 6.5.1 Diluent N,N-Dimethyiformamide 6.5.2 Blank solution Use diluent as blank Pipette 2,0 mL. of the blank into a 20 mL headspace vial and screw with cap. CQA025/F02-01 - Approved By (Signature):Watson ULSD COPY _India Ambernath (Pharma) _ ANALYTICAL METHOD VALIDATION PROTOCOL Product : Validation Protocol No.: SIMVASTATIN USP VPKAD100076 _ Version No:00 Method : WATIARD/RMS/0099(T)-00 Effective date 201 RESIDUAL SOLVENTS BY GAS CHROMATOGRAPHY Page No.: 7 of 23, 6.5.3 6.5.4 6.5.5 6.5.6 Standard stock solution (2000 jig per ml of Methanol, 500 yg per ml of Petroleum ether , 25000 ug per ml of Ethyl acetate and 2000 pg per ml of Tetahydrofuran, 2000 yg per ml of Toluene.) Accurately transfer about 200 mg of Methanol,50 mg of Petroleum ether, 2500 mg of Ethyl acetate, 200 mg of Tetahydrofuran and 200 mg of Toluene into a 100 mL volumetric flask containing about 25 mL of diluent. Dilute and bring to volume with diluent and shake to mix thoroughly. (Prepare in duplicate and Label as standard stock solution-1 and standard stock solution-2) Standard Solution-1 (200 yg per ml of Methanol, 50 ug per ml of | Petroleum ether , 2500 ug per ml of Ethyl acetate and 200 pig per ml of Tetahydrofuran, 200 yg per ml of Toluene.) Pipette 5.0 mL of the standard stock solution-1 into a 50 mL volumetric flask, bring to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL of the Standard solution-1 into a 20 mL headspace vial and screw with cap. Standard Solution-2 (200 yg per ml of Methanol, 50 yg per ml of Petroleum ether , 2500 ug per ml of Ethyl acetate and 200 pg per ml of Tetahydrofuran, 200 pg per'ml of Toluene.) Pipette 5.0 mL of the standard stock solution-2 into a 50 mL volumetric flask, bring to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL of the Standard solution-2 into a 20 mL headspace vial and screw with cap, Sample Solution Accurately weigh and quantitatively transfer about 2.0 g of the sample into a 20 mL headspace vial, add 2.0 mL of diluent, screw with cap and mix well. (Prepare in duplicate) CQA025/F02-04 Approved By (Signature): __ y@as2°— LeWatson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL Validation Protocol No. Product : SIMVASTATIN USP VPKAD100076 m ; Version No:00 . WATIARDIRMS/0099(T)-00__| Effective datey Method : RESIDUAL SOLVENTS BY GAS 16 JUL 2010 CHROMATOGRAPHY Page No.: 8 of 23 66 a7 CQA025/F02-01 6.7.1 The relative standard deviation of peak area response and retention time for 6.7.2. The similarity factor between the responses due to standard solution-1 and Procedure Set up the chromatographic conditions as described under 6.3 and 6.4, and allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Solution No. of injections Blank solution 3 Standard solution: 1 ‘Standard solution-2 6 ‘Sample solution _ 4 Bracket every six injections of sample solution with one injection of standard solution 2. Inject blank wherever required and at the end of set. Record the peak area responses for the major peak and check for system suitability requirements. Proposed System Suitability criteria Test is valid only if following system suitability requirements are met. each solvent from six replicate injections of the standard solution-2 should not be more than 15.0 % and 1.0 % respectively. standard solution-2 for each solvent should be between 0.85 to 1.15. Calculate the similarity factor for each solvent as follows. Peak area response of respective solvent in standard solution (1) x Weight of respective solvent in standard solution (2) Peak area response of respective solvent in standard solution (2) x Weight of respective solvent in standard solution (1) Note: If similarity factor does not fall within 0.85 and 1.15, prepare fresh standard solution in duplicate and reinject. Determine the peak area response and calculate similarity factor as above. oO FT Approved By (Signature): Se |Watson India Ambernath (Pharma) LEN CORY elena sOQUT ANALYTICAL METHOD VALIDATION PROTOCOL Validation Protocol No.: Product : SIMVASTATIN USP VPKAD100076 Version No:00 . WATIARDIRMS/0099(T)-00 _| Effective date: Method : RESIDUAL SOLVENTS BY GAS 16 JUL 201 CHROMATOGRAPHY Page No.: 9 of 23 Elution order for residual solvent Sr. Solvent Elution ord jer | Retention time (mins) Methanol About 3.976 Petroleum Ether Fraction 1 [ About 6.185 Petroleum Ether Fraction 2___| About 7.511 [Petroleum Ether Fraction 3 | About 6.270 Petroleum Ether Fraction 4 | About 9.213 Petroleum Ether Fraction 6 | About 11.321 __| Ethyl acetate About 12.048 [Tetrahydrofuran About 12.544 Petroleum Ether Fraction 7 | About 13.117, 7 14 | Toluene 7 2 3 4 5 6 | Petroleum Ether Fraction 5 _| About 10.774 T 8 9 0 1 ‘About 17.755 6.8 Calculations for content for each residual solvent Calculate the quantity, of each solvent in ppm as follows. | Ru Wt 5 2 ppm of each solvent = x 105 Where, Ru =. Peak area response of the respective solvent in the chromatogram obtained for the sample solution. Rs = _ Average peak area response of the respective solvent in the chromatograms obtained for replicate injections of the standard | solution 2. wt = Weight of corresponding standard in mg, used in the preparation of standard solution 2. Ws = Weight of sample ing. L 40° = Conversion factor CQA025/F02-01 [free By (Signature): wee”Watson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL | 7 Validation Protocol No.: Product: SIMVASTATIN USP VPKAD100076 | — Version No:00 | . WAT/ARD/RMS/0099(T)-00 Effective date: Method : RESIDUAL SOLVENTS BY GAS 16 JuL 2010 CHROMATOGRAPHY Page No.: 10 of 23 Note: Add the content of all seven petroleum ether fractions and then calculate total petroleum ether content. | 7.0 EXPERIMENTAL OUTLINE | In order to validate the proposed gas chromatographic method for the determination of | residual solvents in Simvastatin evaluate the following validation characteristics in accordance with ICH guidelines (Q2R1, Validation of analytical procedures: Methodology, ICH November 2003) and GSOP no. GSOP 022-001 Revision: 00, *Method Validation’ (effective date 25 January 2010). + Specificity | © Quantitation Limit (LOQ) * Linearity and range + Accuracy + Precision > Repeatability > Intermediate precision . Robustness 74 Specificity Perform specificity to ensure that peaks of individual residual solvents are separated from each other and from interference due to blank, if any. 7.1.4 Preparation of solutions 7AA4, Standard stock solution Refer section 6.5 of ‘Method of Analysis’. 7.1.1.2, Standard Solution Refer section 6.5 of ‘Method of Analysis’. i CQA025/F02-01 Approved By (Signature): Gyo —_____Watson India Ambernath (Pharma) _ ANALYTICAL METHOD VALIDATION PROTOCOL PLES Copy | | Validation Protocol No.: Product : SIMVASTATIN USP VPKAD100076 _ Version No:00 WATIARD/RMS/0099(T)-00 Effective date: RESIDUAL SOLVENTS BY GAS 16 Jul 201 CHROMATOGRAPHY Page No.: 11 of 23 Method : 7.1.1.3. Preparation of Identification Solutions 7.4.1.3.1 For Methanol ‘Accurately transfer about 200 mg of Methanol into a 100 mL volumetric flask containing about 2 mL of diluent. Dilute and bring to volume with diluent and shake to mix thoroughly. Pipette 5.0 mL of the above solution into a 50 mL volumetric flask, bring to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL of the above solution into a 20 mL headspace vial and screw with cap. 7.1.1.3.2. For Petroleum ether Accurately transfer about 50 mg of Petroleum ether into a 100 mL volumetric flask containing about 25 mL of diluent. Dilute and bring to volume with diluent and shake to mix thoroughly. Pipette 5.0 mL of the above solution into a 50 mL volumetric flask, bring to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL. of the above solution into a 20 mL headspace vial and screw with cap. 7.1.1.3.3 For Ethyl acetate ‘Accurately transfer about 2500 mg of Ethyl acetate into a 100 mL. volumetric flask containing about 25 mL of diluent. Dilute and bring to volume with diluent and shake to mix thoroughly. Pipette 5.0 mL of the above solution Into a 50 mL volumetric flask, bring to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL of the above solution into a 20 mL headspace vial and screw with cap. CQa025/F02-01 | Approved By (Signature): oro LUWatson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL _ Validation Protocol No.: Product : SIMVASTATIN USP VPKAD100076 _ Version No:00 : WATIARD/RMS/0099(7)-00 Effective date: Method : RESIDUAL SOLVENTS BY GAS a1 6 JUL 20) CHROMATOGRAPHY Page No.: 12 of 23 ~~ 7.4.4.3.4 For Tetahydrofuran Accurately transfer about 200 mg of Tetahydrofuran into a 100 mL volumetric flask containing about 25 mL of diluent. Dilute and bring to volume with diluent and shake to mix thoroughly. Pipette 5.0 mL of the above solution into a 50 mL volumetric flask, bring | to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL of the above solution into a 20 mL headspace vial and screw with cap, 7.4.1.3.5 For Toluene Accurately transfer about 200 mg of Toluene into a 100 mL volumetric flask containing about 25 mL of diluent. Dilute and bring to volume with diluent and shake to mix thoroughly. Pipette 5.0 mL of the above solution into a 50 mL volumetric flask, bring to volume with diluent and shake to mix thoroughly. Pipette 2.0 mL of the above solution into a 20 mL headspace vial and screw with cap. 7.1.1.4, Sample solution ‘Accurately weigh and quantitatively transfer about 2.0 g of the sample into a 20 mL headspace vial, add 2.0 mL of diluent, screw with cap and mix well. 7.1.1.8. Spiked Sample solution ‘Accurately weigh and quantitatively transfer about 2.0 g of the sample into a 20 ml headspace vial, add 2.0 mL of standard solution, screw with cap and mix well, 74.2 Chromatographic system Refer sections 6.3 and 6.4 of ‘Method of Analysis’. CQA025/F02-01Watsor®. SO NUTRAL LET COPY India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL ” ” Validation Protocol No.: Product: | SIMVASTATIN USP VPKAD100076 i Version No:00 WATIARD/RMS/0099(7)-00 | - Effective dat Method : RESIDUAL SOLVENTS BYGAS p16 JUL 7010) CHROMATOGRAPHY Page No.: 13 of 23 74.3 Procedure Set up the chromatographic conditions as described under 6.3 and 6.4, and | allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Inject blank solution in triplicate from different vials. Inject Standard solution-1 once. Inject Standard solution-2 in six replicates from different vials. Inject each Identification solution once. Inject Sample solution once. | Inject Spiked sample solution once. 7.4.4 Acceptance Criteria 7.1.4.1 There should be no interference from the blank solution at the retention time of each residual solvent. If interference is observed inject diluent in three replicates and determine average response. 7.1.42 The average interference should not be greater than 5% with respect to standard solution response of the respective solvent. | 7.2 Quantitation Limit (LOQ) Based on the response obfained for the standard solution (200 pg per mi of Methanol, 50 pg per ml of Petroleum ether , 2500 ug per mi of Ethyl acetate and 200 ug per mi of Tetahydrofuran, 200 ug per ml of Toluene) prepare a solution of an appropriate concentration that would yield an acceptable signal-to-noise ratio (greater than 10) 7.24 Preparation of solutions | 7.2.1.4 Standard stock solution Refer section 6.5 of ‘Method of Analysis’ CQA025/F02-01 i | Approved By (Signature): aeWatson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL | Validation Protocol No.: Product : ‘SIMVASTATIN USP VPKAD100076 Version No:00 . WAT/ARDIRMS/0099(T)-00 _| Effective date: Method: RESIDUAL SOLVENTS BY GAS 6 JUL 2019 Page No.: 14 of 23, | | CHROMATOGRAPHY “7.2.4.2 Standard Solution Refer section 6.5 of ‘Method of Analysis’ 7.2.1.3 LOQ solution Based on the response obtained for the standard solution, prepare a solution of an appropriate concentration that would yield an acceptable signal-to- noise ratio (greater than 10). 7.2.2 Chromatographic system Refer sections 6.3 and 6.4 of ‘Method of Analysis’. 7.2.3 Procedure: Set up the chromatographic conditions as described under 6.3 and 6.4, and allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Inject blank solution in triplicate from different vials. Inject Standard solution-tonce. Inject Standard solution-2 in six replicates from different vials. Inject LOQ solution in six replicates from different vials. Calculate the signal to noise ratio for each solvent peak using the formula, Signal to noise (S/N) ratio = xu ‘Where, H_ = Height of solvent peak from the baseline when a perpendicular is dropped from the apex of the peak in the chromatogram of the LOQ solution. h = Average height of the noise calculated from the chromatogram obtained after injection of blank, observed over a distance equal to at least 20 times the width at half-height of the peak in the CQA025/F02-01 Ne | Approved By (Signature) ee | LeWatson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL ~ ~ ~ Validation Protocol No.: | Product : SIMVASTATIN USP VPKAD100076 | | | _ Version No:00 Method : WATI/ARD/RMS/0099(T)-00 Effective date: 1 6 JUL 201 : RESIDUAL SOLVENTS BY GAS CHROMATOGRAPHY Page No.: 15 of 23 “chromatogram obtained with the LOG solution and situated equally | around the place where this peak would be found. | Using this S/N ratio value evaluate the calculated LOQ 7.2.4 Acceptance Criteria 7.2.4.1 The amount is acceptable as LOQ if the signal-to-noise ratio of residual solvent is equal to, or more than 10. [Ensure that the signal-to-noise ratio is sufficiently greater than 10 so that the concentration is valid as LOQ even | when the method is reproduced on other GC systems in the same laboratory, or in different laboratories. 7.2.4.2 The relative standard deviation of peak area response and retention time for each major peak in six replicate injections of the LOQ solution should not be more than 15.0 % and 1.0 % respectively. 7.3 Linearity and Range Evaluate linearity of each residual solvent at five different concentration levels from LOQ to 120% of the limit level concentration. | 7.3.1. Preparation of solutions 7.3.1.4. Standard stock solution Refer section 6.5 of ‘Method of Analysis’ 7.3.1.2. Standard Solution Refer section 6.5 of ‘Method of Analysis’ 7.3.1.3. Stock solution for linearity levels Use standard stock solution (Solution A) 7.3.1.4. Stock solution for LOQ level (Solution B) Based on the LOQ concentration, prepare a solution of an appropriate concentration for each residual solvent CQA025/FO2-01 | Approved By (Signature):Watson India Ambernath (Pharma) AONTROLLE CORY ANALYTICAL METHOD VALIDATION PROTOCOL, r a " Validation Protocol N Product : SIMVASTATIN USP VPKAD100076 _ : Version No:00 . WATIARDIRMS/0099(T)-00 Effective dat Method : RESIDUAL SOLVENTS BYGAS FTG SUL 2010 CHROMATOGRAPHY Page No.: 16 of 23 7.3.1.8. Solutions for linearity and range Prepare as mentioned in Table 1 below ' 4 25) .251, gine? 3 | 382 382 5 ge seed ben geuleee gah 28| 228) see Sis é g srs se 3) spe 50 | 160.0 | 40.0 | 20000 | 160.0 | +600 3000 | 580 | 25000 | 2000 | 2000 | oe ee oe 30000 | 2400 | zoo | 420 | | ‘O ml of the each Linearity level solution into a separate 20 mL headspace vial and screw with cap. 60 \ 50 * Prepare level 1 by taking appropriate volume of respective standard solutions, based on the required ‘LOQ’ concentration of each solvent 7.3.2. Chromatographic system j Refer sections 6.3 and 6.4 of ‘Method of Analysis’. ) 7.3.3. Procedure Set up the chromatographic conditions as described under 6.3 and 6.4, and | allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Inject blank solution in triplicate from different vials. Inject Standard solution-1 once. Inject Standard solution-2 in six replicates from different vials. Inject the linearity solutions for first level and last level in six replicates and other levels in duplicate and take average peak area response for calculation. Determine the linearity correlation coefficient for each solvent studied | CQA025/FO2-01 Approved By (Signature): eedWatsor™ LERANTAM LER cory India Ambernath (Pharma) us oe ANALYTICAL METHOD VALIDATION PROTOCOL Validation Protocol No.: Product : SIMVASTATIN USP VPKAD100076 _ Version No:00 . WATI/ARD/RMS/0089(T)-00 Effective date: Method : RESIDUAL SOLVENTS BY GAS 416 jul 2010 CHROMATOGRAPHY Page No.: 17 of 23 7.3.4. Acceptance criteria 7.3.4.1. Linearity correlation coefficient of average peak area response of replicate injections plotted against respective concentration, calculated for each solvent should not be less than 0.98. 7.3.4.2. For each solvent, the relative standard deviation of peak area response and retention time for six replicate injections of the first and last linearity level should not be more than 15.0 % and 1.0 % respectively. 7.4 Accuracy (Recovery in %) Evaluate accuracy (recovery in %) at a minimum of 4 levels in the range from LOQ to 120% (LOQ, 50%, 100% and 120%) of the limit level concentration for each solvent. Prepare each level in triplicate and inject once. 7.44. Preparation of solutions 7.4.1.1 Standard stock solution. Refer section 6.5 of ‘Method of Analysis’. 7.4.1.2 Standard solution. Refer section 6.5 of ‘Method of Analysis’ 7.41.3 Stock solution Prepare as standard stock solution (Solution C) 7.4.1.4 Stock solution for LOQ level (Solution D) Based on the LOQ concentration, prepare a solution of an appropriate concentration for each residual solvent T4AS Sample solution Accurately weigh and quantitatively transfer about 2.0 g of the sample into a 20 mL headspace vial, add 2.0 mL. of diluent, screw with cap and mix well. (Prepare in triplicate) CQA025/F02-01Watson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL Product : Validation Protocol No.: 1 ‘SIMVASTATIN USP. VPKAD100076 | _ Version No:00 Method : WATIARDIRMS/0099(7)-00 Effective dat RESIDUAL SOLVENTS BY GAS | 116 JUL 2010 CHROMATOGRAPHY Page No.: 18 of 23 741.6 T4AT TANS TANS 7.41.10 T4414 TAAAZ Standard solution for accuracy at LOQ level (Solution E) Prepare depending on the required LOQ concentration for each residual solvent, Standard solution for accuracy at 50% level (Solution F) Pipette 2.5 mL of Solution C into a 0 mL volumetric flask and bring to volume with diluent and shake to mix thoroughly. Standard solution for accuracy at 100% level (Solution G) Pipette 5 mL of Solution C into a 50 mL volumetric flask and bring to volume | with diluent and shake to mix thoroughly. Standard solution for accuracy at 120% level (Solution H) Pipette 6 mL of Solution C into a 50 mL volumetric flask and bring to volume with diluent and shake to mix thoroughly. | Sample Solution for accuracy at LOQ level Accurately weigh and quantitatively transfer about 2.0 g of the sample into a separate 20 mL headspace vial, add 2.0 mL of Solution E, screw with cap and mix well. (Prepare in triplicate). Sample Solution for accuracy at 50% level ‘Accurately weigh and quantitatively transfer about 2.0 g of the sample into a separate 20 mL headspace vial, add 2.0 mL of Solution F, screw with cap and mix well. (Prepare in triplicate). ‘Sample Solution for accuracy at 100 % level ‘Accurately weigh and quantitatively transfer about 2.0 g of the sample into a separate 20 mL headspace vial, add 2.0 mL of Solution G, screw with cap and mix well, (Prepare in triplicate). CQA025/F02-01 - eH | Approved By (Signature): ase > |Watson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL ] ] Validation Protocol No.: CQA025/F02-01 yo Approved By (Signature): Le j 7.4.2. 7.4.3. 74. 75 7.54 Product : SIMVASTATIN USP | vPKAD100076 LL _— Version No:00 . WATIARDIRMS/0099(T)-00 _| Effective dat Method: | RESIDUAL SOLVENTS BY GAS 16 Jul 201 CHROMATOGRAPHY Page No.: 19 of 23 7.41.13 Sample Solution for accuracy at 120 % level Accurately weigh and quantitatively transfer about 2.0 g of the sample into a separate 20 ml headspace vial, add 2.0 mL of Solution H, screw with cap and mix well. (Prepare in triplicate). Procedure Set up the chromatographic conditions as described under 6.3 and 6.4, and allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Inject blank solution in triplicate from different vials. Inject Standard solution-1 once. Inject Standard solution-2 in six replicates from different vials. Inject Sample solution in triplicate from different vials. | Inject the sample solutions for accuracy levels in triplicate from different vials. Calculate % recovery at each level. Acceptance Criteria 3.1. For each solvent, recovery should be between 80% and 120% for each preparation and 86% to 114% for average at each level. Precision | System Precision and Repeatability To evaluate the repeatability, perform analysis on six weighings of a sample. Calculate the % RSD for each residual solvent obtained for the six sample solutions. If any of the solvent is not detected in the sample or if the peak area response is lower than the peak area response of the respective solvent in LOQ solution, then spike the required solvent in the sample solution at the respective target level concentration.Watson India Ambernatl h (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL Product : Validation Protocol No.: SIMVASTATIN USP VPKAD100076 Version No:00 Method : WATIARD/RMS/0099(T)-00 RESIDUAL SOLVENTS BY GAS CHROMATOGRAPHY 78.14 76144 7.5.1.1.2 75.211 75212 75.1.2 75.13 | _75.1.3.4 CQA025/F02-01 __The relative standard deviation for peak area response and_ retention | Preparation of solutions Standard stock solution Refer section 6.5 of ‘Method of Analysis’ Standard solution Refer sections 6.5 of ‘Method of Analysis’ Sample solution Accurately weigh and quantitatively transfer about 2.0 g of the sample into a 20 mL headspace vial, add 2.0 mL of diluent, screw with cap and mix well Spiked Sample solution ‘Accurately weigh and quantitatively transfer about 2.0 g of the sample into a separate 20 mL headspace vial, add 2.0 mL of standard solution into the Headspace vial, screw with cap and mix well. Procedure Set up the chromatographic conditions as described under 6.3 and 6.4, and allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Inject blank solution in triplicate from different vials. Inject Standard solution-1 once. Inject Standard solution-2 in six replicates from different vials. Inject Sample solution in six replicates from different vials, Inject Spiked sample solution in six replicates from different vials. Acceptance Criteriarangst Watson dia Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL | Validation Protocol No.: | Product : SIMVASTATIN USP VPKAD100076 _ Version No:00 : WAT/ARDIRMS/0099(T)-00 Effective date: | Method: | ReSinUAL SOLVENTS BY.GAS 6 JUL ort | CHROMATOGRAPHY Page No.: 21 of 23 ~ ime for each Solvent peak in six replicate injections of the Standard | Solution-2 should not be more than 15.0 % and 1.0 % respectively. 7.5.1.3.2 For each solvent, the similarity factor between the response due to Standard Solution-1 and Standard Solution-2 should be between 0.85 to | 1.15, 7.5.1.3.3 RSD of the content of each residual solvent obtained in the six preparations of sample should not be more than 15.0%. 7.5.2 Intermediate Precision Perform repeatability as described in section 7.5.1 on a different day and by a different analyst (with all the solutions prepared individually) using a different instrument, Evaluate the results by comparing with those obtained during repeatability. | 7.5.21 Procedure Refer section 7.5.1.2 of ‘Repeatability’ 7.5.2.2 Acceptance criteria 7.5.2.2.1 RSD of the content of each residual solvent obtained in the six preparations of sample solution should not be more than 15.0%. 7.5.2.2.2 The cumulative RSD for the content of each residual solvent along with results obtained for precision study should not be more than 15.0%. 7.6 Robustness Determine the robustness of the proposed method by making deliberate alterations to the chromatographic conditions by minor variations and check the effect of the same on the results obtained. To evaluate the robustness, determine the system suitability criteria at each | change. 7.6.1 Preparation of solutions | __Refer section 7.5.1.1 of ‘Repeatability’. COAo2s/F02-01 SS Approved By (Signature): _ SAPWatson India Ambernath (Pharma) ANALYTICAL METHOD VALIDATION PROTOCOL 1 Validation Protocol No.: Product : | SIMVASTATIN USP VPKAD100076 | _ Version No:00_ | . WAT/ARD/RMS/0099(7)-00 Effective date: | Method : RESIDUAL SOLVENTS BY GAS FG SUL 2010) CHROMATOGRAPHY | Page No.: 22 of 23 76.1.1 Procedure Set up the chromatographic conditions as described under 6.3 and 6.4, and allow the system to get stabilized. Inject the gaseous headspace of the following solutions sequentially, and measure the peak area response. Inject blank solution in triplicate from different vials. Inject Standard solution-1 once. Inject Standard solution-2 in six replicates from different vials. Inject Sample solution once Inject Spiked sampie solution once Inject all the solutions by applying below mentioned deliberate changes in the chromatographic conditions. 7.6.2 Change in the Column lot number Refer sections 6.3 and 6.4 of ‘Method of Analysis’ except for the column lot number to be different 7.6.3 Change in the Carrier gas flow rate by + 15% 7.6.3.1 Carrier gas flow rate: 5.5 mL/min Refer sections 6.3 and 6.4 of ‘Method of Analysis’ except for the Carrier gas flow rate to be set at 5.5 mL/min instead of 6.5 mL/min 7.8.3.2 Carrier gas fiow rate: 7.5 mLImin | Refer sections 6.3 and 6.4 of ‘Method of Analysis’ except for the Carrier gas | flow rate to be set at 7.5 mL/min instead of 6.5 mL/min 7.6.4 Change in the Detector temperature by + 2°C 7.6.4.1 Detector temperature: 262°C Refer sections 6.3 and 6.4 of ‘Method of Analysis’ except for Detector temperature to be set at 262°C instead of 260°C CQA025/F02-01 — — Approved By (Signature) eee —Watson India Ambernath (Pharma) __ ANALYTICAL METHOD VALIDATION PROTOCOL T ~ ~~ Validation Protocol No.: Product: | SIMVASTATIN USP VPKAD100076 __ Version No:00 . WATIARDIRMSI0099(T)-00 __| Effective dat Method : RESIDUAL SOLVENTS BY GAS 16 JUL 20M CHROMATOGRAPHY | Page No.: 23 of 23 7.6.4.2 Detector temperature: 258°C | Refer sections 6.3 and 6.4 of ‘Method of Analysis’ except for Detector temperature to be set at 258°C instead of 260°C 7.6.5 Acceptance Criteria 7.6.5.1 There should be no interference from the blank solution at the retention time of each residual solvent. If interference is observed inject diluent in three replicates and determine average response. 7.6.5.2 The average interference should not be greater than 5% with respect to standard solution response of the respective solvent. 7.6.5.3 For each solvent, the relative standard deviation for peak area response and retention time for six replicate injections of the Standard Solution-2 should not be more than 15.0 % and 1.0 % respectively. 7.6.5.4 For each solvent, the similarity factor between the responses due to Standard Solution-1 and Standard Solution-2 should be between 0.85 to 4.15, | CQA025/F02-01 - Approved By (Signature): eae i L