Ezetimibe VPGQC14061 00 Ya

0 Analytical Method Validation Protocol Goa, India
Watson
Validation Protocol No.: VPGQC14061
Material Name EZETIMIBE Version No.:00
Test Name RESIDUAL SOLVENTS

Effective date: 0 2 JU N 2914
Page 1 of 16
APPROVALS
Prepared by: Reviewed by(QC):
ittixt4y)
I t_R
Sign & Date Sign & Date
per; Sb(11'4/ CO-ci C.c..Ur)

Name Name
Reviewed by (QA): Approved by(QA):
A \ott. (02,,ogetAl•I
Sign & Date
Sign & Date
°LK° AAkit" fc'1 /Iv \ ca rr.\.,--e—)

Name Name
CQA156/F01-01
Watson
e Analytical Method Validation Protocol Goa, India

Material Name EZETIMIBE
Version No.:00
Effective Date 0 2._FL'); 2014
Page 2 of 16
1.0 Purpose: To provide the documentary evidence that, the method of analysis used for residual
solvents in Ezetimibe gives consistent, reliable and reproducible results.
2.0 Scope : This protocol is limited to partial validation of the residual solvents as part of unilateral
method transfer by partial method validation for Ezetimibe.
3.0 Background: The Ezetimibe is currently used in the manufacturing of Ezetimibe and Simvastatin
Tablets by Watson Pharma Pvt. Ltd., Goa, India. This product line was transferred
from Watson Laboratories Inc. Florida to Watson Pharma Pvt. Ltd., Goa, India.
The deficiency letter is received as a Day 70 comments from EU for

Ezetimibe/Simvastatin Actavis 10/10mg, 10/20mq, 10/40mg, 10/80mq tablets
(Reference No. DK/H/2343/001-004/DC). The below query is raised by EU on Drug
Master File for Ezetimibe active Pharmaceutical Ingredient.
Tetrahydrofuran, a class II solvent, is used in stage 3 of the synthesis (referring to

the flow-chart and the presented description). Control of this solvent should be
ensured in an appropriate specification (intermediate or active substance) with a
limit not exceeding the EU/ICH Q3C limit of 720 ppm using a validated analytical
method. The specification revised to include a test/limit for control should be
presented. Batch analysis results should be presented demonstrating compliance
with the set limit of NMT 720 ppm
The vendor Teva has responded to above query as below:

The carry over study of Tetrahydrofuran to Ezetimibe drug substance was
performed testing of THF on three representative production batches. The results
are presented below:
Batch number Tetrahydrofuran

614460110 <R.L (15 ppm)
614460210 <R.L (15 ppm)
614460310 <R.L (15 ppm)
R.L- reported limit
According to the results, the found contents of Tetrahydrofuran in the tested
batches were less than RL (15ppm) which is below 10% of the ICH limit for
Tetrahydrofuran (720ppm). As requested, Ezetimibe monograph for residual
solvent was updated to include a routine test for THE with a limit of not more than
720ppm (as per ICH guideline -class 2 solvent). Vendor has performed analytical
method validation for residual solvent THF. Vendor residual solvent test method
and Watson Goa residual solvent test methods are same. Hence to update current
Watson Goa test method to include residual solvent THF, partial analytical method
validation will be performed at Watson Goa as part of method transfer.
CQA156/F01-01 Approved By:

Watson
Validation Protocol No.:VPGQC14061

Effective Date
0 2. ,,,i, 2014
Page 3 of 16
4.0 Reference:
4.1 Standard operating procedure CQA/156-01 titled "Analytical method validation/verification"
4.2 Watson Goa Standard Test Procedure: RMS/0554-00
4.3 Vendor Teva analytical method ID: IN-6144-RS, Revision No.: 3
5.0 Site of the study: Watson Goa, India
6.0 Responsibility: Analyst / Section Head / Head Quality Control / Quality Assurance.
7.0 Instruments and Materials:

7.1 Instrument: Gas Chromatography
7.2 Standard/solvent:
• Acetone; GC grade or equivalent
• Methanol; GC Grade or equivalent
• 2-propanol; GC Grade or equivalent.
• Methylene chloride; HPLC or equivalent
• Toluene; GC grade or equivalent
• Tetrahydrofuran; GC grade or equivalent
• N, N dimethylformamide; GC grade or equivalent
8.0 Validation Procedure:

Perform the partial method validation by checking the following validation parameters as per
corresponding experimental design.
Validation Parameters:
8.1 Specificity
8.2 Limit of Limit of Quantitation and Limit of Detection
8.3 Accuracy
8.4 Precision (Repeatability and Intermediate precision)
9.0 Acceptance criteria:
Acceptance criteria for above validation parameters are specified in experimental design.
10.0 Results:
10.1 Observation and results to be recorded in method validation data sheet.
10.2 Summarize the findings of the method validation study to draw inference.
11.0 Conclusion:
Based on the interpretation of the results in method validation summary draw the conclusion.

Watson

Effective Date 0 a JUN 2014
Page 4 of 16
12.0 METHOD DETAILS:

12.1 Reagents, Solvents and Standards
• Acetone; GC grade or equivalent

• Methanol; GC Grade or equivalent
• 2-propanol; GC Grade or equivalent.
• Methylene chloride; HPLC or equivalent
• Toluene; GC grade or equivalent
• Tetrahydrofuran (THF); GC grade or equivalent
• N, N dimethylacetamide (DMA); GC grade or equivalent
12.2 Chromatographic system

Instrument : Agilent gas chromatograph or equivalent
Column : DB-624 Column, 30m x 0.53 mm ID x 3.0 pm (Agilent) or equivalent.
Carrier gas : Helium, constant pressure- 3.6 psi (about 5 mUmin at 40°C)
Injection mode : split
Split Ratio : 1:4
Detector : Flame Ionization Detector
Temperature : Injector: 180°C
Detector: 250°C
Oven program : Initial temperature: 40°C; Initial time: 3.0 min
Rate Final temp. Final time
10°C/min. 150°C 2.0 min
Head space parameters:
Vial pressure : 12.5 psi
Temperature : Oven : 90°C
Loop :100°C
Transfer line : 110°C
Times:
GC cycle : 26 min*
Sample eq. : 35 min.
Pressurize : 0.20 min.
Loop fill : 0.10 min.
Loop eq. : 0.05 min.
Injection : 0.50 min.
Shaking : Low
Loop volume : 1 mL
* Note: GC Cycle time is a function of room temperature that may be modified accordingly.

el, Analytical Method Validation Protocol Goa, India
Watson
Effective Date 0 2 JL;,; 2014
Page 5 of 16
12.3 Preparation of the Standard Solution:

12.3.1 Standard Stock Solution:
Weigh accurately about 150 mg of methanol, about 250 mg of 2-Propanol, about 30
mg of methylene chloride, about 250 mg of acetone, about 44.5mg of Toluene and
about 36 mg of Tetrahydrofuran into a 25mL volumetric flask containing about 15mL
N, N-Dimethylacetamide (DMA). Make up to volume with DMA and mix thoroughly
by inverting the flask multiple times.
➢ Prepare stock standard solution in duplicate and name as Stock standard

solution-A and Stock standard solution-B
12.3.2 Standard Solution (3000 ppm of methanol, 5000 ppm of 2-propanol, 600 ppm of
methylene chloride, 5000 ppm of acetone, 890ppm of Toluene and 720 ppm
of THF with respect to test concentration).
Dilute 1 mL Standard Stock Solution to 20 mL with DMA. Transfer 1 mL of this
solution into 20mL headspace vials, immediately seal the vial with a septum crimp cap
and test according to the headspace GC conditions.
➢ Prepare standard solution in duplicate and name as standard solution-A

and standard solution-B
Note: The standard solution is stable for at least five days at room temperature in
transparent bottle.
12.3.3 QL solution(About 25 ppm of methanol, 25 ppm of 2-propanol, 25 ppm of

methylene chloride, 25 ppm of acetone, 10 ppm of Toluene and 15 ppm of
Tetrahydrofuran with respect to test concentration)
Weigh accurately about 250 mg of methanol, 250 mg of 2-Propanol, 250 mg of
methylene chloride, 250 mg of acetone, about 100 mg Toluene and 150 mg of THF
into a 250 mL volumetric flask containing about 150 mL N, N-Dimethylacetamide
(DMA). Make up to volume with DMA and mix thoroughly by inverting the flask
multiple times.
Dilute 1 mL the above solution to 20 mL with DMA and mix thoroughly by inverting
the flask multiple times. (QL stock solution)
Dilute 1 mL of the above solution to 20 mL with DMA and mix thoroughly by inverting
the flask multiple times.
Transfer 1mL of this solution into a 20mL headspace vial. Immediately seal the vial
with a septum crimp cap and test according to the headspace GC conditions.

Watson

Effective Date
0 2 JUN 2014
Page 6 of 16
12.4 Test preparation: (Conc: 100 mg/mL)

Transfer about 100 mg accurately weighed sample into a 20mL headspace vial. Add 1 mL
of N, N-Dimethylacetamide.
Immediately seal with a septum crimp cap and mix gently and test according to the headspace
GC conditions.
Note: The sample solution stored in vial at room temperature is stable at least for
two days.
12.5 Procedure:
12.5.1 Sequence of injection
Note: Inject one injection per vial
Table:1 Sequence of Injection
Solution Replicates
Diluent 3
Tentative QL solution 3
Diluent 1
Standard solution A 1
Standard solution B 3
Diluent 1
Test preparation 1
Tentative QL solution 1
Bracketing Standard 1
Continue the sequence of Bracketing with QL solution and Standard Solution B after
every six injections of Test preparation and at the end of sequence.
12.5.2 System Suitability Requirements

12.5.2.1 The % RSD values for each replicate response factors (Response factor =
respective solvent peak area in standard solution/concentration of the
respective solvent in standard solution in pg/mL) and for all six response
factors of each individual peak responses from the standard solution B and
tentative QL solution should be:
Table-2: %RSD requirement

Standard B and
Solvent Name Standard B QL solution
QL solution
Methanol Not more than 5.0% Not more than 15.0% Not more than 15.0%
Acetone Not more than 5.0% Not more than 15.0% Not more than 15.0%
2-Propanol Not more than 5.0% Not more than 15.0% Not more than 15.0%
Methylene chloride Not more than 10.0% Not more than 15.0% Not more than 15.0%
Toluene Not morethan10.0% Not more than 15.0% Not more than 15.0%
THE Not morethan10.0% Not more than 15.0% Not more than 15.0%

Analytical Method Validation Protocol Goa, India
0
Watson

Effective Date 0 2 JUi 2014
Page 7 of 16
12.5.2.2 The theoretical plates for the toluene peak from the standard solution B
should be not less than 70000
12.5.2.3 The similarity factor between two Standard Solutions i.e. Standard Solution-A
and Standard Solution-B should be
Table-3: Similarity factor
Solvent Name Similarity factor
Methanol 0.95 to 1.05
Acetone 0.95 to 1.05
2-Propanol 0.95 to 1.05
Methylene chloride 0.90 to 1.10
Toluene 0.90 to 1.10
Tetrahydrofuran 0.90 to 1.10
12.6 Calculation:
12.6.1 Calculate the similarity factor as follows
Area of respective Solvent Weight of respective Standard

from Standard Solution A from Standard Stock Solution B (mg)
Area of respective Solvent Weight of respective Standard

from Standard Solution B from Standard Stock Solution A (mg)
12.6.2 Calculate the Content of particular solvent as follows:
ru Ws 1(mL) 1(mL) P(%)

ppm of residual solvent- x x x x X106
rs 25(m L) 20(m L) Wu 100
Where,
ru = Peak area of each residual solvent from the test solution.
rs = Mean peak area of each residual solvent from the standard solution B
Wu = Weight of sample, in mg
Ws = Weight of each residual solvent taken in standard stock solution B (mg).
P(%) = Purity of respective standard solvent
12.7 Specification:
Methanol NMT 3000 ppm
Acetone NMT 5000 ppm
2-Propanol NMT 5000 ppm
Methylene chloride : NMT 600 ppm
Toluene : NMT 890 ppm
Tetrahydrofuran : NMT 720 ppm

Watson
Effective Date 0 2 j,). 3 2614
Page 8 of 16
13.0 EXPERIMENTAL DESIGN

13.1 SPECIFICITY AND SYSTEM SUITABILITY:
Reagents, Solvents and Standards, Chromatographic system and Preparation of solution,
standard solution, refer method details.
13.1.1 Preparation of Identification solution:
13.1.1.1 Preparation of Methanol ID solution
Weigh accurately about 150 mg of methanol, into a 25mL volumetric flask
containing about 15mL N, N-Dimethylacetamide (DMA). Make up to volume with
DMA and mix thoroughly by inverting the flask multiple times.
Dilute 1 mL above solution to 20 mL with DMA.
Transfer 1 mL of this solution into 20mL headspace vials, immediately seal the
vial with a septum crimp cap and test according to the headspace GC conditions.
13.1.1.2 Preparation of Acetone ID solution

Weigh accurately about 250 mg of Acetone, into a 25mL volumetric flask
13.1.1.3 Preparation of 2-Propanol ID solution

Weigh accurately about 250 mg of 2-Propanol, into a 25mL volumetric flask
13.1.1.4 Preparation of Methylene chloride ID solution

Weigh accurately about 30 mg of methylene chloride, into a 25mL volumetric
flask containing about 15mL N, N-Dimethylacetamide (DMA). Make up to
volume with DMA and mix thoroughly by inverting the flask multiple times.
13.1.1.5 Preparation of Toluene ID solution

Weigh accurately about 44.5 mg of Toluene, into a 25mL volumetric flask

Watson

Effective Date 0 2 JUN 2014
Page 9 of 16
13.1.1.6 Preparation of Tetrahydrofuran ID solution

Weigh accurately about 36 mg of Tetrahydrofuran, into a 25mL volumetric flask
13.1.3 Preparation of sample solution:

Sample (As such):
of N, N-Dimethylacetamide.
Immediately seal with a septum crimp cap and mix gently and test according to the
headspace GC conditions.
Sample (Mock)
of standard solution-B.
13.1.4 Procedure:
Sequence of injection:
1) Diluent(triplicate) 2) Individual ID solutions 3)Tentative Quantitation solution(Three replicate) 4)
standard solution-A 5) standard solution-B(three replicate) 6) Diluent (triplicate) 7) Sample solution 8)
Mock sample solution 9) Tentative Quantitation solution 10) Brac. standard solution-B
13.1.5 Acceptance criteria:

The interference from other sources is NMT 5%.
Resolution between any two peaks must be more than 1.5.
The theoretical plates for the toluene peak from the standard solution B should be
3 not less than 70000.
4 Reproducibility for peak area of solvents as per table 2 for Tetrahydrofuran only.
5 Similarity factor as per the table 3 for Tetrahydrofuran only.

Watson

Version No.:00
Effective Date 0 Z _./.. r 2014
Page 10 of 16
13.2 LIMIT OF QUANTITATION (LOQ) AND LIMIT OF DETECTION (LOD):

13.2.1 Preparation of LOD solution:

Different concentration of solvents may be used as required that meets the LOD requirement
and will be reported in the final report as LOD concentration.
13.2.2 Preparation of LOQ solution:
Separate LOQ will not be performed as Quantitative solution injection is part of test method.
Same solution will be injected six replicate.
13.2.3 Procedure:
1) Diluent 2) Tentative Quantitation solution(six replicate) 3) standard solution-A 4) standard
solution-B(three replicate) 5) Diluent 6) Tentative Quantitation solution 7) Brac. standard solution-B
I The theoretical plates for the toluene peak from the standard solution B should be
not less than 70000.
Reproducibility for peak area of solvents as per table 2 for Tetrahydrofuran only.
I Similarity factor as per the table 3 for Tetrahydrofuran only.
Signal-to-noise ratio for Limit of quantitation for THE should be more than or
4
equal to 10:1.
Signal-to-noise ratio for Limit of detection for the THE should be more than or
5
equal to 3:1.

et Analytical Method Validation Protocol Goa, India
Watson
Effective Date 0 Z 1,,, 2J14
Page 11 of 16
13.4 ACCURACY
13.4.1 Preparation of accuracy levels:

Weigh accurately about 36 mg of Tetrahydrofuran, into a 25 mL volumetric flask containing
about 15mL N, N-Dimethylacetamide (DMA). Make up to volume with DMA and mix
thoroughly by inverting the flask multiple times.
Table 4: Accuracy dilutions
Sr. Volume of Accuracy Diluted to Concentration of Concentration in
No. stock solution to be mL with THF in ppm % of each THF
taken DMA
1 1.0* 20 1.5 2
2 0.5 20 36 50
3 1.0 20 72 100
4 1.5 20 108 150
* To be pipetted from QL stock solution (Section 12.3.3)
13.4.2 Preparation of Accuracy solution:

Level-1
of Level 1 as per table 4.
(Note : Prepare samples in three replicates.)
Level-2
Level-3

Watson
o Analytical Method Validation Protocol Goa, India

Effective Date t. — G14
.
T st Name RESIDUAL SOLVENTS e
Page 12 of 16
Level-4
13.4.3 Procedure:
1) Diluent 2) Tentative Quantitation solution(six replicate) 3) standard solution-A 4) standard
solution-B(three replicate) 5) Diluent 6) Accuracy Level-1(Three preparation) 7) Accuracy Level-2 (Three
preparation) 8) Accuracy Level-3 (Three preparation) 9) Accuracy Level-4(Three preparation)
Inject bracketing standard after every six injection of sample solution.
1 not less than 70000.
Accuracy should be 80% to 120% for each preparation. 86% to 114% for average
4 at each levels.

Watson

Effective Date 0 2 j, : :
Test Name RESIDUAL SOLVENTS `' 2014
Page 13 of 16
13.5 PRECISION
13.5.1 REPEATABILITY:
Reagents, Solvents and Standards, Chromatographic system and Preparation of
solution, standard solution refer method details.
13.5.1.1 System precision

Evaluate system precision by injecting six replicate injection of standard solution-
B.
13.5.1.2 Method precision

To evaluate the method precision, perform analysis on six different preparations
of sample. Calculate `)/0 RSD of content obtained for the six sample solutions.
Sample solution
Transfer about 100 mg accurately weighed sample into a 20mL headspace vial.
Add 1 mL of N, N-Dimethylacetamide.
(Prepare in six replicate)
(Note: If THF will not detected in the method precision spike THF at limit level concentration as
applicable. This experiment can be combined with the accuracy study by preparing six samples
at 100% level).
Spiked sample:
Add 1 mL of standard solution-B.
13.5.1.3 Procedure
1) Diluent 2) Quantitation solution(six replicate) 3) standard solution-A 4) standard
solution-B(three replicate) 5) Diluent 6) Sample-1 7) Sample-2 8) Sample-3 9)
Sample-4 10) Sample-5 11) Sample-6 12) Brac. standard solution-B

Watson
Effective Date 0 2. J,,. .- 2014
Page 14 of 16
13.5.1.4 Acceptance criteria:

The theoretical plates for the toluene peak from the standard solution B should be I
1
not less than 70000.
4 The %RSD for content of THE for six sample preparation should not be more than
j 15 %.

Watson
Version No.:00

Effective Date 0 a JL,. v 2014
Page 15 of 16
13.5.2 INTERMEDIATE PRECISION:

Reagents, Solvents and Standards, Chromatographic system and Preparation of
solution, standard solution refer method details.
Repeat the repeatability experiment on second day performed by second analyst by using
another instrument.
13.5.2.1 System precision
Evaluate system precision by injecting six replicate injection of standard solution-

B.
13.5.2.2 Method precision

To evaluate the method precision, perform analysis on six different preparations
of sample. Calculate % RSD of content obtained for the six sample solutions.
Sample solution
Add 1 mL of N, N-Dimethylacetamide.
(Note: If THF will not detected in the method precision spike THF at limit level concentration as
applicable.)
Spiked sample:
Add 1 mL of standard solution-B.
13.5.2.3 Procedure
1) Diluent 2) Tentative Quantitation solution(six replicate) 3) standard solution-A 4) standard solution-
B(three replicate) 5) Diluent 6) Sample-1 7) Sample-2 8) Sample-3 9) Sample-4 10)
Sample-5 11) Sample-6 12) Brac. standard solution-B

Watson
Effective Date 0 a •J 2014
Page 16 of 16
13.5.2.4 Acceptance criteria:

1
not less than 70000
3i Similarity factor as per the table 3 for Tetrahydrofuran only.
The %RSD for content of THE for six sample preparation should not be more than
41 0
%.
1 5 Cumulative %RSD for both repeatability and intermediate precision should not be
_more than 15.0 %.
14.0 CHANGE HISTORY
Change control no. Supersedes Effective date Summary of Changes
GOA/C-14/330-QC None 02 !„: ?014 New Protocol

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0 Analytical Method Validation Protocol Goa, India

Test Name RESIDUAL SOLVENTS

Prepared by: Reviewed by(QC):

per; Sb(11'4/ CO-ci C.c..Ur)

Reviewed by (QA): Approved by(QA):

°LK° AAkit" fc'1 /Iv \ ca rr.\.,--e—)

Validation Protocol No.: VPGQC14061

The deficiency letter is received as a Day 70 comments from EU for

Tetrahydrofuran, a class II solvent, is used in stage 3 of the synthesis (referring to

The vendor Teva has responded to above query as below:

Batch number Tetrahydrofuran

CQA156/F01-01 Approved By:

Test Name RESIDUAL SOLVENTS

7.0 Instruments and Materials:

8.0 Validation Procedure:

9.0 Acceptance criteria:

10.1 Observation and results to be recorded in method validation data sheet.

CQA156/F01-01 Approved By:

Validation Protocol No.:VPGQC14061

12.0 METHOD DETAILS:

• Acetone; GC grade or equivalent

12.2 Chromatographic system

CQA156/F01-01 Approved By:

12.3 Preparation of the Standard Solution:

➢ Prepare stock standard solution in duplicate and name as Stock standard

➢ Prepare standard solution in duplicate and name as standard solution-A

12.3.3 QL solution(About 25 ppm of methanol, 25 ppm of 2-propanol, 25 ppm of

CQA156/F01-01 Approved By:

Test Name RESIDUAL SOLVENTS

12.4 Test preparation: (Conc: 100 mg/mL)

12.5.2 System Suitability Requirements

Table-2: %RSD requirement

CQA156/F01-01 Approved By:

Test Name RESIDUAL SOLVENTS

Area of respective Solvent Weight of respective Standard

Area of respective Solvent Weight of respective Standard

12.6.2 Calculate the Content of particular solvent as follows:

ru Ws 1(mL) 1(mL) P(%)

CQA156/F01-01 Approved By:

13.0 EXPERIMENTAL DESIGN

13.1.1.2 Preparation of Acetone ID solution

13.1.1.3 Preparation of 2-Propanol ID solution

13.1.1.4 Preparation of Methylene chloride ID solution

13.1.1.5 Preparation of Toluene ID solution

CQA156/F01-01 Approved By:

Test Name RESIDUAL SOLVENTS

13.1.1.6 Preparation of Tetrahydrofuran ID solution

13.1.3 Preparation of sample solution:

13.1.5 Acceptance criteria:

CQA156/F01-01 Approved By:

Validation Protocol No.:VPGQC14061

13.2 LIMIT OF QUANTITATION (LOQ) AND LIMIT OF DETECTION (LOD):

13.2.1 Preparation of LOD solution:

13.2.4 Acceptance criteria:

CQA156/F01-01 Approved By:

13.4.1 Preparation of accuracy levels:

* To be pipetted from QL stock solution (Section 12.3.3)

13.4.2 Preparation of Accuracy solution:

CQA156/F01-01 Approved By:

Validation Protocol No.:VPGQC14061

Inject bracketing standard after every six injection of sample solution.

13.4.4 Acceptance criteria:

CQA156/F01-01 Approved By: