QC Assay Validation

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product: SARCIP-500 TABLET

QC/VLD/01

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009

Revision No. : 00

OBJECTIVE : The establish documented evidence which will provide a high assurance that the analytical method for assay of Ciprofloxacin 500 Tablet will give results with acceptable Accuracy, Repeatibility, Relative Standard Deviation & Ruggedness when the Sarcip-500 Tablet. SCOPE :

01-06

degree of in SarcipLinearity, applied to

This method applies to the procedure which is used for calculation and assay of Ciprofloxacin.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product: SARCIP-500 TABLET

QC/VLD/01

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009

METHOD

02-06

Revision No. : 00

:

1.

Weight accurately equivalent to about 100mg of Ciprofloxacin working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with 0.1NHydrochloric Acid and make up to same, mix and filter if required.

2.

With above dilution prepare standard and sample solution of 100mcg/ml with 0.1N Hydrochloric acid separately, mix and make homogeneous solution.

3.

Pipette 3ml,5ml,7ml,9ml and 11ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.

4.

To each flask add 1ml of 5%w\v solution of Anhydrous Ferric chloride in water. Make up with distilled water to produced 50ml, mix to make homogeneous solution.

5.

Measure the absorbance at maximum at about 438nm,against water as blank and tabulate the observed data in the observation table.

6.

Plot linearity curve from the different concentration corresponding area of Ciprofloxacin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

7.

Plot repeatability curve from by taking Ciprofloxacin solution serial no. on X-axis and observed peak area on Y-axis.

8.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

9.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product: SARCIP-500 TABLET

QC/VLD/01

Prepared By

03-06

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Ciprofloxacin in Sarcip-500 Tables shall be studied by measuring the absorbance of the standard Ciprofloxacin solution of 20 – 100mcg/ml concentration & range 438nm. Linearity curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Ciprofloxacin in mcg\ml

Peak area

1.

20mcg\ml

405

2.

40mcg\ml

450

3.

60mcg\ml

535

4.

80mcg\ml

570

5.

100mcg\ml

605

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/01

04-06

SARCIP-500 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Ciprofloxacin in Sarcip-500 Tables shall be studied by measuring the absorbance of the standard Ciprofloxacin solution of 100mcg/ml concentration & range 438nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY Sr. No.

Concentration in mcg\ml

Peak area

1

100mcg\ml

604

2.

100mcg\ml

608

3.

100mcg\ml

603

4.

100mcg\ml

605

5.

100mcg\ml

604

Sarthak Biotech Pvt. Ltd.

Total Page:

05-06

Assay Method Validation Name of the Product: SARCIP-500 TABLET Prepared By

QC/VLD/01

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION Test concentration of Ciprofloxacin used = 100mcg\ml. Number of test solution used = 5 604+608+603+605+604 Average area = ----------------------------------5 Dev. S =

= 604.8

(604-604.8)2 +(608-604.8)2 +(603-604.8)2 +(605-604.8)2 +(604-604.8)2 [ --------------------------------------------------------------------------------------] 1/2 5-1 0.64+10.24+3.24+0.04+0.64 = [--------------------------------------]1/2 4 14.8 = [ -------]1/2 4 = [ 3.7]

1/2

= 1.9235

RSD

1.9235 x 100 = -----------------604.8

Limit

: ( Not more than 2.0% )

= 0.318%

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/01

06-06

SARCIP-500 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Ciprofloxacin in Sarcip-500 tablet shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days.

RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

0.1168 0.1156 0.1158

0.1348 0.1352 0.1342

Measured Absorbance at of Ref. Std.

Measured Absorbance at 438nm of sample

% of Ciprofloxacin

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 438nm of sample

% of Ciprofloxacin

602 607 605

608 612 615

100.11 99.61 101.35

0.1162 0.1165 0.1164

0.1349 0.1356 0.1352

606 609 612

601 616 608

99.43 101.15 99.55

Average wt of tablet 670.2 mg Mean 100.35% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.2 % Standard Deviation : ±10 %

Averagewt.of tablet Mean Standard Deviation

675.1 mg 100.04% 10%

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/02

01-06

SARFLOK-200 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

PURPOSE :-The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Ofloxacin in Sarflok-200 Tablet will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarflok-200 Tablet. SCOPE:-

This method applies to the procedure which is used for calculation and assay of Ofloxacin.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS :1. Linearity. 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/02

02-06

SARFLOK-200 TABLET

Prepared By Date of Issue/Effective Date : 18/11/2007

Checked By/ Approved By Review Date : 17/11/2009

Revision No. : 00

METHOD 1.

Weigh accurately equivalent to about 50mg of Ofloxacin working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with 0.1NHydrochloric Acid and make up to same, mix and filter if required.

2.

Pipette 3ml,4ml,5ml,6ml and 7ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.

4.

To each flask add 1ml of 5%w\v solution of Ferric Chloride. Make up the volume with distilled water to produce 50ml, mix to make homogeneous solution.

5.

Measure the absorbance at maximum at about 437nm,against water as blank and calculate the observed data in the observation table.

6.

Plot linearity curve from the different concentration corresponding area of Ofloxacin peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.

7.

Plot repeatability curve from by taking Ofloxacin solution serial no. on X-axis and observed peak area on Y-axis.

8.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

9.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/02

03-06

SARFLOK-200 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Ofloxacin in Sarflox-200 Tables shall be studied by measuring the absorbance of the standard Ofloxacin solution of 30 – 70mcg/ml concentration & range 437nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No.

Concentration of Ofloxacin in mcg\ml

Peak area

1.

30mcg\ml

368

2.

40mcg\ml

437

3.

50mcg\ml

512

4.

60mcg\ml

591

5.

70mcg\ml

664

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/02

04-06

SARFLOK-200 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Ofloxacin in Sarflox-200 Tables shall be studied by measuring the absorbance of the standard Ofloxacin solution of 50 mcg/ml concentration & range 437nm.Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY S.No.

Concentration in mcg\ml

Peak area

1

50mcg\ml

515

2.

50mcg\ml

512

3.

50mcg\ml

515

4.

50mcg\ml

509

5.

50mcg\ml

519

Sarthak Biotech Pvt. Ltd.

Total Page:

05-06

Assay Method Validation Name of the Product:

QC/VLD/02

SARFLOK-200 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION Test concentration of Ofloxacin used = 50mcg\ml. Number of test solution used = 5 515 + 512 + 515 + 509 + 519 Average area = -----------------------------------------5

Dev. S =

= 513.4

(515-513.4)2 +(512-513.4)2 +(515-513.4)2 +(509-513.4)2 +(519-513.4)2 1/2 [ ---------------------------------------------------------------------------------------] 5-1

2.56+1.96+1.96+19.36+31.36 = [---------------------------------------]1/2 4 57.2 = [ -------] 1/2 4 = [ 14.3]

RSD

Limit

1/2

3.781 x 100 = -----------------513.4

= 3.781

= 0.736%

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

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06-06

Assay Method Validation Name of the Product:

QC/VLD/02

SARFLOK-200 TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 18/11/2007

Review Date : 17/11/2009

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Ofloxacin in Sarflok-200 tablet shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

% of Ofloxacin

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbanc e at of Ref. Std.

Measured Absorbance at 437nm of sample

% of Ofloxacin

0.0498 0.0501 0.0512

0.1021 0.1019 0.1029

100.67 101.7 100.7

0.0499 0.0512 0.0489

0.1006 0.1018 0.1012

512 504 508

505 510 514

98.97 102.95 98.90

Measured Measured Absorbanc Absorbance e at at 437nm of Ref. of sample Std.

509 511 519

512 515 512

Average wt of tablet 410.4 mg Mean 101.02% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.65 % Standard Deviation : ±10 %

Averagewt.of tablet Mean Standard Deviation

404.6 mg 100.27% 10%

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

SARNIM-P TABLET

01-11

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 19/12/2007

Review Date : 18/12/2009

Revision No. : 00

PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Nimesulide & Paracetamol in Sarnim-P Tablet will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarnim-P Tablet. SCOPE :

This method applies to the procedure which is used for calculation and assay of Paracetamol & Nimesulide.

RESPONSIBILITY :

1. Quality Control Manager. 2. Asstt. Quality Control Chemist.

VALIDATION PARAMETERS: 1.

Linearity.

2.

Repeatibility.

3.

Relative Standard Deviation.

4.

Ruggedness.

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

SARNIM-P TABLET

02-11

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 19/12/2007

PROCEDURE (For Paracetamol)

Review Date : 18/12/2009

Revision No. : 00

:

1.

Weight accurately equivalent to about 60mg of Paracetamol working Reference standard and its equivalent to sample in a 100ml volumetric flask . Dissolve with 20ml of Hydrochloric Acid 1M and digest for 30 minutes, then make up the volume 100ml with distilled water.

2.

Pipette 2ml,3ml,4ml,5ml and 6ml of standard and sample solution separately in a calibrated 50ml. measuring flask.

3.

To each flask add 2ml of 10%w\v solution of Sodium Nitrate & allow to stand for 5 minutes, then add 2ml of 10% w/v solution of Ammonium Sulphamate. Again allow to stand for 5 to 6 minutes now further add 2 ml. of 10%w/v solution of Sodium Hydroxide and keep it for 5 minutes. Now adjust the volume to 50ml with distilled water.

4.

Measure the absorbance at maximum at about 450nm, against water as blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Paracetamol peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Paracetamol solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

SARNIM-P TABLET

03-11

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 19/12/2007

Review Date : 18/12/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Paracetamol in Sarnim-P Tables shall be studied by measuring the absorbance of the standard Sarnim-P solution of 24 – 72mcg/ml concentration & range 450nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No.

Concentration of Paracetamol in mcg\ml

Peak area

1.

24mcg\ml

525

2.

36mcg\ml

595

3.

48mcg\ml

645

4.

60mcg\ml

695

5.

72mcg\ml

742

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/03

SARNIM-P TABLET

04-11

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 19/12/2007

Review Date : 18/12/2009

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Paracetamol in Sarnim-P Tables shall be studied by measuring the absorbance of the standard Sarnim-P solution of 24 mcg/ml concentration & range 450nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

24mcg\ml

525

2.

24mcg\ml

528

3.

24mcg\ml

527

4.

24mcg\ml

526

5.

24mcg\ml

525

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

SARNIM-P TABLET

05-11

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 19/12/2007

Review Date : 18/12/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION Test concentration of Paracetamol used = 24mcg\ml. Number of test solution used = 5 525 + 528 + 527 + 526 + 525 Average area = -----------------------------------------5

Dev. S =

= 526.2

(525-526.2)2 +(528-526.2)2 +(527-526.2)2 +(526-526.2)2 +(525-526-2)2 1/2 [ ---------------------------------------------------------------------------------------] 5-1

1.44+3.24 +0.64+0.04+1.44 = [---------------------------------------]1/2 4 6.8 = [ -------]1/2 4 = [ 1.7]

RSD

Limit

1/2

=

1.303 x 100 = -----------------526.2

1.303

= 0.2477%

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

SARNIM-P TABLET

06-11

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 19/12/2007

Review Date : 18/12/2009

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Paracetamol in Sarnim-P tablet shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

0.0598 0.0601 0.0592

0.0784 0.0792 0.0784

Measured Absorbance at of Ref. Std.

Measured Absorbance at 450nm of sample

% of Paraceta mol-mol

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 450nm of sample

% of Paracetamol

521 532 518

528 528 525

100.83 98.23 99.19

0.0602 0.0592 0.0612

0.0782 0.0779 0.0791

512 519 522

518 515 525

101.99 98.75 101.9

Average wt of tablet 652.2 mg Mean 99.41% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.15 % Standard Deviation : ±10 %

Averagewt.of tablet Mean Standard Deviation

654.8 mg 100.88% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/03

07-11

SARNIM-P TABLET

Prepared By Date of Issue/Effective Date : 19/12/2007

Checked By/ Approved By Review Date : 18/12/2009

Revision No. : 00

PROCEDURE (For Nimesulide)

:

1.

Weight accurately equivalent to about 50mg of Nimesulide working Reference standard and its equivalent to sample in a 100ml volumetric flask . Dissolve with 30 to 40ml of 0.1M solution of Sodium Hydroxide. Digest both the solution for 10 minutes then make up the volume to 100ml with 0.1M solution of Sodium Hydroxide.

2.

Pipette 2ml,3ml,4ml,5ml and 6ml of standard and sample solution separately in a calibrated 50ml.measuring flask.

3.

To each flask add 20ml 0.1M Sodium Hydroxide solution shake well and further adjust the volume up to 50ml with 0.1M solution of Sodium Hydroxide.

4.

Measure the absorbance of both the solution at 405nm, against 0.1M Sodium Hydroxide solution as blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Nimesulide peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Nimesulide solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/03

08-11

SARNIM-P TABLET

Prepared By Date of Issue/Effective Date : 19/12/2007

Checked By/ Approved By Review Date : 18/12/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Nimesulide in Sarnim-P Tablets shall be studied by measuring the absorbance of the standard Nimesulide solution of 20 – 60mcg/ml concentration & range 405nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No.

Concentration of Nimesulide in mcg\ml

Peak area

1.

20mcg\ml

552

2.

30mcg\ml

595

3.

40mcg\ml

635

4.

50mcg\ml

702

5.

60mcg\ml

757

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/03

09-11

SARNIM-P TABLET

Prepared By Date of Issue/Effective Date : 19/12/2007

Checked By/ Approved By Review Date : 18/12/2009

Revision No. : 00

REPEATBILITY STUDY Repeatibility assay method of Nimesulide in Sarnim-P Tablets shall be studied by measuring the absorbance of the standard Nimesulide solution of 20 mcg/ml concentration & range 405nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

20mcg\ml

559

2.

20mcg\ml

561

3.

20mcg\ml

561

4.

20mcg\ml

560

5.

20mcg\ml

563

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

10-11

SARNIM-P TABLET

Prepared By Date of Issue/Effective Date : 19/12/2007

Checked By/ Approved By Review Date : 18/12/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION

Test concentration of Nimesulide used = 20mcg\ml. Number of test solution used = 5 559 + 561 + 561 + 560 + 563 Average area = -----------------------------------------5

Dev. S =

= 560.8

(559-560.8)2 +(561-560.8)2 +(561-560.8)2 +(560-560.8)2 +(563-560.8)2 1/2 [ ---------------------------------------------------------------------------------------] 5-1

3.24+0.04 +0.04+0.64+4.84 = [---------------------------------------]1/2 4 8.8 = [ -------]1/2 4 = [ 2.2] RSD

Limit

1/2

=

1.4142 x 100 = -----------------560.8 :

1.4142

= 0.2521%

( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/03

Prepared By

SARNIM-P TABLET Checked By/ Approved By

11-11

Date of Issue/Effective Date : 19/12/2007

Review Date : 18/12/2009

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Nimesulide in Sarnim-P tablet shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

0.0499 0.0489 0.0501

0.3258 0.3251 0.3251

Measured Absorbance at of Ref. Std.

Measured Absorbance at 405nm of sample

% of Nimesuli de

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 405nm of sample

% of Nimesuli de

560 542 560

568 545 555

101.31 98.64 99.61

0.0498 0.0496 0.0502

0.3261 0.3269 0.3251

565 559 553

560 562 556

99.11 99.88 101.65

Average wt of tablet 652.2 mg Mean 99.85% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.03 % Standard Deviation : ±10 %

Averagewt.of tablet Mean Standard Deviation

654.8 mg 100.21% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/04

01-06

SARFIX-100 DT TABLET

Prepared By Date of Issue/Effective Date : 28/11/2007

Checked By/ Approved By Review Date : 27/11/2009

Revision No. : 00

PURPOSE :- The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Cefixime Trihydrate in Sarfix100 DT Tablet will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Sarfix-100 DT Tablet. SCOPE:-

This method applies to the procedure which is used for calculation and assay of Cefixime Trihydrate.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS :1. Linearity. 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Total Page:

Sarthak Biotech Pvt. Ltd. Assay Method Validation Name of the Product:

02-06

QC/VLD/04

SARFIX-100 DT TABLET

Prepared By Date of Issue/Effective Date : 28/11/2007

Checked By/ Approved By Review Date : 27/11/2009

Revision No. : 00

METHOD 1.

Weigh accurately equivalent to about 100mg of Cefixime working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with Methanol and make up to volume up to 100ml.

2.

Pipette 1ml,2ml,3ml,4ml and 5ml of standard and sample solution separately in a calibrated 100ml.volumetric flask.

3.

To each flask add Methanol and make up the volume up to 100ml.

4.

Measure the absorbance at maximum at about 268nm,against Methanol as blank and calculate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Cefixime peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Cefixime solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/04

03-06

SARFIX-100 DT TABLET

Prepared By Date of Issue/Effective Date : 28/11/2007

Checked By/ Approved By Review Date : 27/11/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Cefixime Trihydrate in Sarfix-100 DT Tablets shall be studied by measuring the absorbance of the standard Cefixime Trihydrate solution of 20 – 50mcg/ml concentration & range 268nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No.

Concentration of Cefixime in mcg\ml

Peak area

1.

10mcg\ml

495

2.

20mcg\ml

515

3.

30mcg\ml

540

4.

40mcg\ml

570

5.

50mcg\ml

595

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/04

04-06

SARFIX-100 DT TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 28/11/2007

Review Date : 27/11/2009

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Cefixime Trihydrate in Sarfix-100 DT Tablets shall be studied by measuring the absorbance of the standard Cefixime Trihydrate solution of 10mcg/ml concentration & range 268nm. Repeatibility curve shall be plotted for absorbance against concentration OBSERVATION TABLE FOR REPEATIBILITY S.No.

Concentration in mcg\ml

Peak area

1

10mcg\ml

495

2.

10mcg\ml

492

3.

10mcg\ml

497

4.

10mcg\ml

495

5.

10mcg\ml

496

Sarthak Biotech Pvt. Ltd. Assay Method Validation Name of the Product:

Total Page:

05-06

QC/VLD/04

SARFIX-100 DT TABLET

Prepared By Date of Issue/Effective Date : 28/11/2007

Checked By/ Approved By Review Date : 27/11/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION Test concentration of Cefixime used = 10mcg\ml. Number of test solution used = 5 495 + 492 + 497 + 495 + 496 Average area = -----------------------------------------5

Dev. S =

= 495

(495-495)2 +(495-492)2 +(495-497)2 +(495-495)2 +(495-496)2 [ --------------------------------------------------------------------------] 5-1

1/2

0+9+4+0+1 = [---------------------------------------]1/2 4 14 = [ -------] 1/2 4 = [ 3.5]

1/2

= 1.8708

RSD

Limit

1.8708 x 100 = -----------------495

= 0.377%

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/04

06-06

SARFIX-100 DT TABLET

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 28/11/2007

Review Date : 27/11/2009

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Cefixime Trihydrate in Sarfix-100 DT Tablet shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

0.1128 0.1119 0.1131

0.4019 0.4021 0.4012

Measured Absorbance at of Ref. Std.

Measured Absorbance at 268nm of sample

% of Cefixime

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 268nm of sample

% of Cefixime

498 492 498

502 499 488

101.75 101.50 99.34

0.1118 0.1129 0.1125

0.4028 0.4031 0.4022

502 499 492

508 505 498

101.21 102.14 102.02

Average wt of tablet 402.8 mg Mean 100.86% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 101.32 % Standard Deviation : ±10 %

Averagewt.of tablet Mean Standard Deviation

403.6 mg 101.79% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/05

SARMOX-500 CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/10/2007

Review Date : 09/10/2009

PURPOSE:

01-06

Revision No. : 00

The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Amoxycillin in Sarmox-500 Capsule will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarmox-500 Capsule.

SCOPE :

This method is applies to the procedure which is used for calculation and assay of Amoxycillin.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist.

VALIDATION PARAMETERS: 1. Linearity. 2. Repeatability. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/05

SARMOX-500 CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/10/2007 METHOD 1.

02-06

Review Date : 09/10/2009

Revision No. : 00

:

Weight accurately 100mg Reference standard and equivalent to 100mg. Amoxycillin as test sample. Bowl it with 20 to 30ml of distilled water and make up to produce 100ml.

2.

Pipette 2ml, 3ml, 4ml, 5ml and 6ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.

3.

In a 50ml volumetric flask pipette 2ml Reference and test solution separately. To each flask add 2ml of Phosphate buffer pH 9.2 and add 1ml Vitamin B6 dye..

4.

Measure the absorbance without delay at maximum at about 605nm,against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Amoxycillin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Amoxycillin solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/05

03-06

SARMOX-500 CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/10/2007

Review Date : 09/10/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.

Linearity of assay method of Amoxycillin in Sarmox-500 Capsule shall be studied by measuring the absorbance of the standard Amoxycillin solution of 40 – 120mcg/ml concentration & range 605nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Amoxycillin in mcg/ml

Peak area of

1.

40mcg\ml

652

2.

60mcg\ml

695

3.

80mcg\ml

728

4.

100mcg\ml

765

5.

120mcg\ml

805

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/05

04-06

SARMOX-500 CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/10/2007

Review Date : 09/10/2009

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Amoxycillin in Sarmox-500 Capsule shall be studied by measuring the absorbance of the standard Amoxycillin solution of 40mcg/ml concentration & range 605nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATBILITY Sr.No. 1

Concentration in mcg\ml 40mcg\ml

Peak area 651

2.

40mcg\ml

653

3.

40mcg\ml

652

4.

40mcg\ml

657

5.

40mcg\ml

651

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/05

05-06

SARMOX-500 CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/10/2007

Review Date : 09/10/2009

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Amoxycillin used = 40mcg\ml. Number of test solution used = 5 651+653+652+657+651 Average area = ----------------------------------5 Dev. S =

= 652.8

(651-652.8)2 +(653-652.8)2 +(652-652.8)2 +(657-652.8)2 +(651-652.8)2 [ -------------------------------------------------------------------------------------] 1/2 5-1 3.24+0.04+0.64+17.64+3.24 = [-------------------------------------]1/2 4 24.8 = [ -------]1/2 4 = [ 6.2 ]

1/2

= 2.489

RSD

Limit

2.489 x 100 = -----------------652.8

= 0.381%

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/05

06-06

SARMOX-500 CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/10/2007

Review Date : 09/10/2009

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Amoxycillin in Sarmox-500 Capsule shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

0.1162 0.1159 0.1165

0.1148 0.1162 0.1155

Measured Absorbance at of Ref. Std.

Measured Absorbance at 605nm of sample

% of Amoxyci -llin

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 605nm of sample

% of Amoxycil -lin

655 654 653

652 658 657

100.79 100.38 101.51

0.1154 0.1162 0.1152

0.1168 0.1169 0.1162

659 652 656

655 662 650

98.57 101.30 98.60

Average fill weight 580.2 mg Mean 100.89% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.19 % Standard Deviation : ±10 %

Average fill weight Mean Standard Deviation

582.2 mg 99.49% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/06

ACIGONE CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date :27/09/2007

PURPOSE:

01-06

Review Date : 26/09/2009

Revision No. : 00

The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Omeprazole in Acigone Capsule will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Acigone Capsule.

SCOPE :

This method is applies to the procedure which is used for calculation and assay of Omeprazole.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity. 2. Repeatability. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/06

ACIGONE CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date :27/09/2007 METHOD 1.

02-06

Review Date : 26/09/2009

Revision No. : 00

:

Weight accurately 20mg Reference standard and equivalent to 20mg. Omeprazole as test sample. Dissolve it with 20 to 30ml of 0.1M Sodium Hydroxide solution shake vigorously 4-5 minutes and make up the volume up to 50ml with 0.1M NaOH solution.

2.

Pipette 1ml, 2ml, 3ml, 4ml and 5ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.

3.

In a 50ml volumetric flask pipette 1ml Reference and test solution separately and make up the volume up to 50ml. with 0.1M Sodium Hydroxide solution.

4.

Measure the absorbance without delay at maximum at about 268nm,against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Omeprazole peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Omeprazole solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/06

03-06

ACIGONE CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date :27/09/2007

Review Date : 26/09/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.

Linearity of assay method of Omeprazole in Acigone Capsule shall be studied by measuring the absorbance of the standard Omeprazole solution of 8 – 40mcg/ml concentration & range 268nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Amoxycillin in mcg\ml

Peak area of

1.

8mcg\ml

397

2.

16mcg\ml

421

3.

24mcg\ml

495

4.

32mcg\ml

565

5.

40mcg\ml

605

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/06

04-06

ACIGONE CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date :27/09/2007

Review Date : 26/09/2009

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Omeprazole in Acigone Capsule shall be studied by measuring the absorbance of the standard Omeprazole solution of 8 mcg/ml concentration & range 268nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY

Sr.No. 1

Concentration in mcg\ml 8mcg\ml

Peak area 398

2.

8mcg\ml

401

3.

8mcg\ml

397

4.

8mcg\ml

399

5.

8mcg\ml

398

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/06

05-06

ACIGONE CAPSULE

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date :27/09/2007

Review Date : 26/09/2009

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Amoxycillin used = 8mcg\ml. Number of test solution used = 5 398+401+397+399+398 Average area = ----------------------------------5 Dev. S =

= 398.6

(398-398.6)2 +(401-398.6)2 +(397-398.6)2 +(399-398.6)2 +(398-398.6)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.36+5.76 +2.56+0.16+0.36 = [---------------------------------------]1/2 4 9.2 = [ -------]1/2 4 = [ 2.3 ]

1/2

= 1.5165 RSD

Limit

1.5165 x 100 = -----------------398.6

= 0.3804%

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/06

06-06

ACIGONE CAPSULE

Prepared By Date of Issue/Effective Date :27/09/2007

Checked By/ Approved By Review Date : 26/09/2009

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Omeprazole in Acigone Capsule shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Sample gm.

0.2657 0.2673 0.2661

0.2940 0.2927 0.2939

Measured Absorbance at of Ref. Std.

Measured Absorbance at 268nm of sample

% of Omepraz -ole

Wt. of Ref. Standard gm.

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 268nm of sample

% of Omeprazole

395 401 393

399 396 396

100.40 99.20 100.38

0.2672 0.2657 0.2673

0.2926 0.2961 0.2926

403 398 396

399 401 393

99.48 99.47 99.75

Average fill weight 293.4 mg Mean 99.99% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 99.78

Average fill weight Mean Standard Deviation

293.4 mg 99.57% 10%

Standard Deviation : ±10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/07

SARMOX DRY SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 29/10/2007

PURPOSE:

01-06

Review Date : 28/10/2009

Revision No. : 00

The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Amoxycillin in Sarmox Dry Syrup will give results with acceptable Accuracy, Linearity, Repeatability,

Relative Standard Deviation & Ruggedness when applied to the Sarmox Dry Syrup. SCOPE :

This method is applies to the procedure which is used for calculation and assay of Amoxycillin.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity. 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/07

SARMOX DRY SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 29/10/2007 METHOD 1.

02-06

Review Date : 28/10/2009

Revision No. : 00

:

Weight accurately 100mg Reference standard and equivalent to 100mg. Amoxycillin as test sample. Bowl it with 20 to 30ml of distilled water and make up to produce 100ml.

2.

Pipette 2ml, 4ml, 5ml, 6ml and 7ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.

3.

In a 50ml volumetric flask pipette 2ml Reference and test solution separately. To each flask add 2ml of Phosphate buffer pH 9.2 and add 1ml Vitamin B6 dye. And make up the volume up to 50ml with water.

4.

Measure the absorbance without delay at maximum at about 605nm,against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Amoxycillin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Amoxycillin solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/07

03-06

SARMOX DRY SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 29/10/2007

Review Date : 28/10/2009

LINEARITY STUDY

Revision No. : 00

The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Amoxycillin in Sarmox Dry Syrup shall be studied by measuring the absorbance of the standard Amoxycillin in solution of 40 – 120mcg/ml concentration & range 605nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Amoxycillin in mcg\ml

Peak area of

1.

40mcg\ml

651

2.

60mcg\ml

695

3.

80mcg\ml

726

4.

100mcg\ml

775

5.

120mcg\ml

804

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/07

04-06

SARMOX DRY SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 29/10/2007

Review Date : 28/10/2009

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Amoxycillin in Sarmox Dry Syrup shall be studied by measuring the absorbance of the standard Amoxycillin in solution of 40 mcg/ml concentration & range 605nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY Sr.No. 1

Concentration in mcg\ml 40mcg\ml

Peak area 651

2.

40mcg\ml

652

3.

40mcg\ml

657

4.

40mcg\ml

660

5.

40mcg\ml

655

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/07

05-06

SARMOX DRY SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 29/10/2007

Review Date : 28/10/2009

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Amoxycillin used = 40mcg\ml. Number of test solution used = 5 651+652+657+660+655 Average area = ----------------------------------5 Dev. S =

= 655

(651-655)2 +(652-655)2 +(657-655)2 +(660-655)2 +(655-655)2 [ ------------------------------------------------------------------------------] 1/2 5-1 16+ 9+ 4 + 25 + 0 = [---------------------------------------]1/2 4 54 = [ -------]1/2 4 = [ 13.5 ]

1/2

= 3.6742

RSD Limit

3.6742 x 100 = -----------------655 : ( Not more than 2.0% )

= 0.560%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/07

06-06

SARMOX DRY SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 29/10/2007

Review Date : 28/10/2009

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Amoxycillin in Sarmox Dry Syrup shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Measure Sample d ml Absorbance at of Ref. Std.

Measured Absorbance at 605nm of sample

% of Amoxycillin

Wt. of Ref. Standard gm.

Wt. of Sample ml

Measured Absorbance at of Ref. Std.

Measured Absorbance at 605nm of sample

% of Amoxycillin

01

0.1161

4ml

653

656

100.54

0.1159

4ml

657

652

99.15

02

0.1168

4ml

658

665

101.76

0.1162

4ml

659

663

100.18

03

0.1155

4ml

652

656

100.82

0.1155

4ml

657

654

99.11

Label Claim

125mg Amoxycillin/ 5ml

Label Claim

125mg Amoxycillin/ 5ml

Mean

100.82%

Mean

100.01%

Standard Deviation

10%

Standard Deviation

10%

Combining mean of I & II of both the analysts : Mean : 100.41 % Standard Deviation : ±10 %

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/08

01-06

CEFSAR-125 DRY SYRUP

Prepared By Date of Issue/Effective Date : 17/01/2008

Checked By/ Approved By Review Date : 16/01/2010

Revision No. : 00

PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Cephalexin in Cefasar-125 Dry Syrup will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Cefasar-125 Dry Syrup. SCOPE :

This method is applies to the procedure which is used for calculation and assay of Cephalexin.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity; 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/08

02-06

CEFSAR-125 DRY SYRUP

Prepared By Date of Issue/Effective Date : 17/01/2008

Checked By/ Approved By Review Date : 16/01/2010

Revision No. : 00

METHOD

:

1.

Weight accurately equivalent 250mg of Cephalexin and dissolve in 20ml of Neutral Methanol and titrate with 0.1M Sodium Hydroxide using Bromomethyl blue as indicator, perform a blank for necessary correction. Each ml. of 0.1M Sodium Hydroxide is equivalent to 0.03474gm. of Cephalexin.

2.

Weight 250mg, 300mg, 350mg., 400mg. & 450mg. Cephalexin in 20ml. of Neutral Methanol & titrate with 0.1M NaOH using Bromomethyl blue as indicator. Each ml. of 0.1m Sodium Hydroxide is equivalent to 0.03474gm. of Cephalexin.

3.

Note the volume consumed & tabulate the observed data in observation table.

4.

Plot linearity curve from the different concentration corresponding area of Cephalexin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

5.

Plot repeatability curve from by taking Cephalexin solution serial no. on X-axis and observed peak area on Y-axis.

6.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

7.

Calculated the result by calculation volume consumed by the Cephalexin.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/08

03-06

CEFSAR-125 DRY SYRUP

Prepared By Date of Issue/Effective Date : 17/01/2008

Checked By/ Approved By Review Date : 16/01/2010

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Cephalexin in Cefsar-125 Dry Syrup shall be studied by 250mgto 450 mg Cephalexin. OBSERVATION TABLE FOR LINEARITY ( Molarity of 0.1M NaOH is 0.102305) Sr.No.

Weight of Cephalexin in mg

Volume consumed

1.

250mg.

7ml

2.

300mg.

8.6ml

3.

350mg.

9.7ml

4.

400mg.

11.2ml

5.

450mg.

12.7ml

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/08

04-06

CEFSAR-125 DRY SYRUP

Prepared By Date of Issue/Effective Date : 17/01/2008

Checked By/ Approved By Review Date : 16/01/2010

Revision No. : 00

REPEATIBILITY STUDY

Repeatibility of assay method of Cephalexin in Cefsar-125 Dry Syrup shall be studied by 250 mg Cephalexin.

OBSERVATION TABLE FOR REPEATIBILITY ( Molarity of 0.1M NaOH is 0.102305) Sr.No.

Weight of Cephalexin

Volume consumed

1

250mg.

7ml

2.

250mg.

6.9ml

3.

250mg.

7.1ml

4.

250mg.

7.2ml

5.

250mg.

6.9ml

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/08

05-06

CEFSAR-125 DRY SYRUP

Prepared By Date of Issue/Effective Date : 17/01/2008

Checked By/ Approved By Review Date : 16/01/2010

Revision No. : 00

RELATIVE STANDARD DEVIATION Test concentration of Cephalexin used = 250mg. Number of test solution used = 5 7+6.9+7.1+7.2+6.9 Average volume = ----------------------------------5

Dev. S =

= 7.02

(7- 7.02)2 +(6.9-7.02)2 +(7.1-7.02)2 +(7.2-7.02)2 +(6.9-7.02)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.0004+0.0144+0.0064+0.0324+0.0144 = [------------------------------------------------------]1/2 4 0.068 = [ -------] 1/2 4 = [ 0.017 ]

1/2

= 0.130

RSD

Limit

0.130 x 100 = -----------------7.02

= 1.857%

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd. Assay Method Validation Name of the Product:

Total Page:

06-06

QC/VLD/08

CEFSAR-125 DRY SYRUP

Prepared By Date of Issue/Effective Date : 17/01/2008

Checked By/ Approved By Review Date : 16/01/2010

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Cephalexin inCefsar-125 Dry Syrup shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : Burette 50ml (Borosil) Location : Chemical Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : Burette 50ml (Borosil) Location : Chemical Lab.

Wt. of Sample

Molarity of 0.1M NaOH

Final Volume used

Assay

Wt. of Sample

Molarity of 0.1M NaOH

Final Volume used

Assay

10ml 10ml 10ml

0.102305 0.102305 0.102305

7.0ml 6.9ml 7.0ml

99.51 98.09 99.51

10ml 10ml 10ml

0.102305 0.102305 0.102305

7.1ml 7.0ml 7.0ml

100.94 99.51 99.52

Label Claim 125mg Cephalexin/5ml Label Claim Mean 99.04% Mean Standard Deviation 10% Standard Deviation Combining mean of I & II of both the analysts : Mean : 99.52% Standard Deviation : ±10%

125mg Cephalexin/5ml 99.99% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/09

01-05

CHLORTEX MOUTH WASH

Prepared By Date of Issue/Effective Date : 10/01/2008

Checked By/ Approved By Review Date : 09/01/2010

Revision No. : 00

PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Chlorhexidine Gluconate in

Chlortex Mouth Wash will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Cefasar-125 Dry Syrup. SCOPE :

This method is applies to the procedure which is used for calculation and assay of Chlorhexidine Gluconate.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity. 2. Repeatability. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/09

CHLORTEX MOUTH WASH

Prepared By Date of Issue/Effective Date : 10/01/2008 METHOD :

02-05

Checked By/ Approved By Review Date : 09/01/2010

Revision No. : 00

1.

Weigh accurately equivalent of 200mg. of standard and dissolve in 100ml of distill water.

Chlorhexidine Gluconate Reference

2.

Further pipette 5ml, 10ml, 15ml, 20ml & 25ml of standard and dilute to 100ml. with water.

3.

Weigh accurately equivalent to 5mg., 10mg., 15mg., 20mg. & 25mg. of sample, basified with 10M NaOH and extracted with Chloroform (2 x 20ml). Combine Chloroform layer and dilute to 50ml with water. Further prepare dilution of 50mcg / ml. with water.

4.

To 1 ml. of each above standard and sample dilutions add 3ml of dye solution and 5ml. of Chloroform. Shake well for 3 to 4 minutes.

5.

Measure the absorbance without delay at maximum at about 410nm, against respective blank and tabulate the observed data in the observation table.

6.

Plot linearity curve from the different concentration corresponding area of Chlorhexidine Gluconate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

7.

Plot repeatability curve from by taking Chlorhexidine Gluconate solution serial no. on X-axis and observed peak area on Y-axis.

8.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

9.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/09

03-05

CHLORTEX MOUTH WASH

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/01/2008

Review Date : 09/01/2010

LINEARITY STUDY

Revision No. : 00

The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Chlorhexidine Gluconate in Chlortex Mouthwash shall be studied by measuring the absorbance of the standard Chlorhexidine Gluconate in solution of 100 – 500mcg/ml concentration & range 410nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Chlorhexidine Gluconate in mcg\ml

Peak area of

1.

100mcg\ml

546

2.

200mcg\ml

642

3.

300mcg\ml

726

4.

400mcg\ml

791

5.

500mcg\ml

805

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/09

04-06

CHLORTEX MOUTH WASH

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/01/2008

Review Date : 09/01/2010

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Chlorhexidine Gluconate in Chlortex Mouthwash shall be studied by measuring the absorbance of the standard Chlorhexidine Gluconate in solution of 100 mcg/ml concentration & range 410nm. Repeatibility curve shall be plotted for absorbance against concentration OBSERVATION TABLE FOR REPEATIBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

100mcg\ml

544

2.

100mcg\ml

545

3.

100mcg\ml

548

4.

100mcg\ml

549

5.

100mcg\ml

546

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/09

05-06

CHLORTEX MOUTH WASH

Prepared By Date of Issue/Effective Date : 10/01/2008

Checked By/ Approved By Review Date : 09/01/2010

Revision No. : 00

RELATIVE STANDARD DEVIATION

Test concentration of Chlorhexidine Gluconate used = 100mcg\ml. Number of test solution used = 5 544+545+548+549+546 Average area = ----------------------------------5

Dev. S =

= 546.4

(544-546.4)2 +(545-546.4)2 +(548-546.4)2 +(549-546.4)2 +(546-546.4)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 5.76+2.005+3.2 +1.99 +0.8 = [---------------------------------------]1/2 4 13.755 = [ ----- ----]1/2 4 = [ 3.439 ] 1/2

= 1.854 RSD

Limit

1.854 x 100 = -----------------546.4

= 0.339 %

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/09

06-06

CHLORTEX MOUTH WASH

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 10/01/2008

Review Date : 09/01/2010

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Chlorhexidine Gluconate in Chlortex Mouthwash shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

% of Chlorhexi dine Gluconate

Wt. of Ref. Standard gm.

Wt. of Sample ml

Measured Absorbance at of Ref. Std.

Measur ed Absorbance at 410nm of sample

% of Chlorhexid ine Gluconate

100.49 99.04 99.69

0.5090 0.4992 0.4984

50ml 50ml 50ml

548 542 542

547 545 545

99.99 100.39 100.23

0.4998 0.4989 0.5012

Label Claim

Wt. of Measure Measured Sample d Absorml Absor- bance at bance 410nm of at sample of Ref. Std.

50ml 50ml 50ml

544 546 543

545 542 545

0.2% Chlorhexidine

Gluconate Mean 99.74% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 99.98 % Standard Deviation : ±10 %

Label Claim

0.2% Chlorhexidine

Mean Standard Deviation

Gluconate 100.02% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

01-16

BRONSYP SYRUP

Prepared By Date of Issue/Effective Date : 24/01/2008

Checked By/ Approved By Review Date : 23/01/2010

Revision No. : 00

PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Salbutamol Sulphate, Bromhexine Hcl & Guaiphensin in Bronsyp Syrup will give results with

acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Bronsyp Syrup. SCOPE :

This method is applies to the procedure which is used for calculation and assay of Salbutamol Sulphate, Bromhexine Hcl & Guaiphensin.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity. 2. Repeatability. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

BRONSYP SYRUP

Prepared By Date of Issue/Effective Date : 24/01/2008

METHOD : (Salbutamol Sulphate)

02-16

Checked By/ Approved By Review Date : 23/01/2010

Revision No. : 00

1.

Weigh accurately 100mg. of Salbutamol Sulphate as Reference standard and dissolve in 100 ml of water and prepare the dilution 100mcg/ml. in water.

2.

Accurately weigh quantity of sample is extracted with water to get 100mcg./ml in respect of Salbutamol Sulphate.

3.

Pipette 3ml, 4ml, 5ml, 6ml & 7ml of standard and sample solution and add 3ml of Sodium Carbonate solution, 0.2ml of P-aminophenol reagent and volume is adjusted to 10ml with water.

4.

Measure the absorbance without delay at maximum at about 635nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Salbutamol Sulphate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Salbutamol Sulphate solution serial no. on Xaxis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

03-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

LINEARITY STUDY

Revision No. : 00

The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Salbutamol Sulphate in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Salbutamol Sulphate in solution of 30 – 70mcg/ml concentration & range 635nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Salbutamol Sulphate in mcg\ml

Peak area of

1.

30mcg\ml

695

2.

40mcg\ml

739

3.

50mcg\ml

795

4.

60mcg\ml

829

5.

70mcg\ml

888

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

04-16

BRONSYP SYRUP

Prepared By Date of Issue/Effective Date : 24/01/2008

Checked By/ Approved By Review Date : 23/01/2010

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Salbutamol Sulphate in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Salbutamol Sulphate in solution of 30 mcg/ml concentration & range 635nm. Repeatibility curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR REPEATIBILITY S.No.

Concentration in mcg\ml

Peak area

1

30mcg\ml

698

2.

30mcg\ml

700

3.

30mcg\ml

697

4.

30mcg\ml

695

5.

30mcg\ml

703

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

05-16

BRONSYP SYRUP

Prepared By Date of Issue/Effective Date : 24/01/2008

Checked By/ Approved By Review Date : 23/01/2010

Revision No. : 00

RELATIVE STANDARD DEVIATION

Test concentration of Salbutamol Sulphate used = 30mcg\ml. Number of test solution used = 5 698+700+697+695+703 Average area = ----------------------------------5

Dev. S =

= 698.6

(698-698.6)2 +(700-698.6)2 +(697-698.6)2 +(695-698.6)2 +(703-698.6)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.36+ 1.96+ 2.56+ 12.96 + 19.36 = [-------------------------------------------]1/2 4 37.2 = [ ----- ----]1/2 4 = [ 9.3 ]

1/2

= 3.049 RSD

3.049 x 100 = -----------------698.6

Limit

: ( Not more than 2.0% )

= 0.436 %

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

06-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Salbutamol Sulphate in Bronsyp Syrup shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

0.1209 0.1216 0.1206

Wt. of Measure Sample d ml Absorbance at of Ref. Std.

5ml 5ml 5ml

698 694 695

Measured Absorbance at 635nm of sample

% of Salbutam ol

Wt. of Ref. Standard gm.

Wt. of Sample ml

Measured Absorbance at of Ref. Std.

Measured Absorbance at 635nm of sample

% of Salbutam ol

702 697 698

100.32 100.76 99.77

0.1218 0.1206 0.1218

5ml 5ml 5ml

695 694 698

694 694 701

100.35 99.22 100.93

Label Claim 2mg. / 5ml Mean 100.28% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.16% Standard Deviation : ±10 %

Label Claim Mean Standard Deviation

2 mg. / 5ml 100.93% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

07-16

BRONSYP SYRUP

Prepared By Date of Issue/Effective Date : 24/01/2008

Checked By/ Approved By Review Date : 23/01/2010

Revision No. : 00

METHOD : (Bromhexine Hcl) 1.

Weigh accurately 40mg. of Bromhexine Hcl. and transfer to a separating funnel, basify with 5ml of dilute Ammonia solution , add 25ml. of water and extract with for 25ml

portions of Chloroform layer through anhydrous Sodium Sulphate previously washed with Chloroform. Evaporate Chloroform layer and dissolve residue in 0.1M Methanolic Hydrochloric Acid to produce 100ml. (400mcg./ ml.). Further make the dilution with Methanolic Hydrochloric Acid to get final concentration of 40mcg/ml. 2.

Measure accurately sample equivalent to 4mg. of Bromhexine Hcl., add 25ml. water and continue with the procedure as described under preparation of standard solution.

3.

Pipette 2ml, 3ml, 4ml, 5ml & 6ml of standard and sample solution, add 8ml of P-dimethyl-aminobenzaldehyde (10%w/v in Methanol) & 5ml. of Hydrochloric Acid mix and allow the stand for 10 minutes and then make up the volume to 25ml. with water.

4.

Measure the absorbance at maximum at about 425nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Bromhexine Hcl peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Bromhexine Hcl. solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

08-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.

Linearity of assay method of Bromhexine Hcl in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Bromhexine Hcl in solution of 3.2 – 9.6mcg/ml concentration & range 425nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Bromhexine Hcl. in mcg\ml

Peak area of

1.

3.2mcg\ml

545

2.

4.8mcg\ml

598

3.

6.4mcg\ml

642

4.

8.0mcg\ml

691

5.

9.6mcg\ml

732

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

09-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Bromhexine Hcl in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Bromhexine Hcl in solution of 3.2mcg/ml concentration & range 425nm. Repeatibility curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR REPEATIBILITY Sr. No.

Concentration in mcg\ml

Peak area

1

3.2mcg\ml

545

2.

3.2mcg\ml

547

3.

3.2mcg\ml

548

4.

3.2mcg\ml

545

5.

3.2mcg\ml

542

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

10-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Bromhexine Hcl. used = 3.2mcg\ml. Number of test solution used = 5

545+547+548+545+542 Average area = ----------------------------------5 Dev. S =

= 545.4

(545-545.4)2 +(547-545.4 )2 +(548-545.4)2 +(545-545.4)2 +(542-545.4)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.16+2.56+6.76+0.16+11.56 = [--------------------------------------------]1/2 4 21.2 = [ ----- ----]1/2 4 = [ 5.3 ]

1/2

= 2.302 RSD

Limit

2.302 x 100 = -----------------545.4

= 0.422 %

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

11-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Bromhexine Hcl in Bronsyp Syrup shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Measure Sample d ml Absorbance at of Ref. Std.

0.0398 0.0396 0.0399

5ml 5ml 5ml

548 544 543

Measured Absorbance at 425nm of sample

% of Bromhex ine

Wt. of Ref. Standard gm.

Wt. of Sample ml

Measured Absorbance at of Ref. Std.

Measured Absorbance at 425nm of sample

% of Bromhexi ne

546 548 545

99.13 99.72 100.11

0.0396 0.0399 0.0401

5ml 5ml 5ml

545 541 542

542 543 543

98.45 100.11 100.43

Label Claim 4mg. Bromhexine/5ml Mean 99.65% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 99.66 Standard Deviation : ±10%

Label Claim Mean Standard Deviation

4mg. Bromhexine/5ml 99.66% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

12-16

BRONSYP SYRUP

Prepared By Date of Issue/Effective Date : 24/01/2008

Checked By/ Approved By Review Date : 23/01/2010

Revision No. : 00

METHOD : (Guaiphenesin) 1.

Weigh accurately 100mg. of Guaiphenesin, transfer to a separating funnel. Add 20ml of water and 5ml. of Sodium hydroxide solution (20%w/v). Extract with for 40ml portions of petroleum ether (60-80oC) . Discard ether layer, collect the aqueous layer and extract

for 20ml portions of chloroform. Pass combined chloroform layer throught anhydrous sodium Sulphate and make up to 100ml with chloroform. 2.

Measure accurately sample equivalent to 100mg. of substance, add 20ml of water and continue the procedure as described under preparation of standard solution.

3.

Pipette 5ml, 6ml, 7ml, 8ml & 10ml of standard and sample solution, and make up the volume 100ml. with methanol.

4.

Measure the absorbance at maximum at about 275nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Guaiphenesin peak. Taking concentration on X-axis and corresponding peak on Yaxis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Guaiphenesin solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

13-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

LINEARITY STUDY

Revision No. : 00

The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Guaiphenesin in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Guaiphenesin in solution of 50 – 100mcg/ml concentration & range 275nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Guaiphenesin in mcg\ml 1.

Peak area of

50mcg\ml

419

2.

60mcg\ml

460

3.

70mcg\ml

502

4.

80mcg\ml

542

5.

100mcg\ml

605

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

14-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Guaiphenesin in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Guaiphenesin in solution of 50 mcg/ml concentration & range 275nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY

Sr.No.

Concentration in mcg\ml

Peak area

1

50mcg\ml

421

2.

50mcg\ml

419

3.

50mcg\ml

424

4.

50mcg\ml

422

5.

50mcg\ml

420

Total Page:

Sarthak Biotech Pvt. Ltd. Assay Method Validation Name of the Product:

15-16

QC/VLD/10

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Guaiphensin used = 50mcg\ml. Number of test solution used = 5

421+419+424+422+420 Average area = ----------------------------------5

Dev. S =

= 421.2

(421-421.2)2 +(419-421.2)2 +(424-421.2)2 +(422-421.2)2 +(420-421.2)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.04+4.84+ 7.84 + 0.64 + 1.44 = [--------------------------------------------]1/2 4 14.8 = [ ----- ----]1/2 4 = [ 3.7 ]

1/2

= 1.924 RSD

Limit

1.924 x 100 = -----------------421.2 :

= 0.457 %

( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/10

16-16

BRONSYP SYRUP

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 24/01/2008

Review Date : 23/01/2010

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Guaiphensin in Bronsyp Syrup shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

Analyst- I (Satpal Saini) Analyst- II (Rajnish Kohli) Date of analysis : Date of analysis : Instrument & Model : Instrument & Model : UV Spectrophotometer, Systronics UV Spectrophotometer, Systronics 117 117 Location : Instrument Lab. Location : Instrument Lab. Wt. of Ref. Standard gm.

Wt. of Measure Sample d ml Absorbance at of Ref. Std.

Measured Absorbance at 275nm of sample

% of Guaiph enesin

Wt. of Ref. Standard gm.

Wt. of Sample ml

Measured Absorbance at of Ref. Std.

Measured Absorbance at 275nm of sample

% of Guaiphen esin

01 02 03

Label Claim 50mg Guaiphenesin/5ml Label Claim Mean Mean Standard Deviation 10% Standard Deviation Combining mean of I & II of both the analysts : Mean : Standard Deviation : ±10 %

50mg Guaiphenesin/5ml 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

01-16

BECLOSAR-NC CREAM

Prepared By Date of Issue/Effective Date : 08/02/2008

Checked By/ Approved By Review Date : 07/02/2010

Revision No. : 00

PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Beclomethasone Dipropionate, Neomycin Sulphate & Clotrimazole in Beclosar-NC Cream will give results

with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Beclosar-NC Cream. SCOPE :

This method is applies to the procedure which is used for calculation and assay of Beclomethasone Dipropionate, Neomycin Sulphate & Clotrimazole.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist. VALIDATION PARAMETERS: 1. Linearity. 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

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Assay Method Validation Name of the Product:

QC/VLD/11

BECLOSAR-NC CREAM

Prepared By Date of Issue/Effective Date : 08/02/2008 METHOD : (Beclomethasone Dipropionate)

02-16

Checked By/ Approved By Review Date : 07/02/2010

Revision No. : 00

1.

Weigh accurately 50mg. of Beclomethasone Dipropionate and dissolve in 100ml. of methanol. Further dilute 1ml. and make up the volume to 50ml. with methanol. This makes 10mcg./ml dilution of Beclomethasone Dipropionate.

2.

Weight accurately sample (cream) equivalent to 0.5mg of Beclomethasone Dipropionate and dissolve in 25ml. of methanol. Filter and then make up the volume to 50ml. with methanol.

3.

Pipette 1ml, 2ml, 3ml, 4ml & 5ml of standard and sample solution, add 6ml. of 0.1M tetrabutyl ammonium hydroxide (TBAH) reagent and 2ml. of 0.5%w/v of Tetrazolium blue (BTZ) solution. Heat the solution on water bath for 15minutes, cool the solution and add 10ml. Glacial Acetic acid.

4.

Measure the absorbance without delay at maximum at about 515nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Beclomethasone Dipropionate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Beclomethasone Dipropionate solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

03-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

LINEARITY STUDY

Revision No. : 00

The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Beclomethasone Dipropionate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Beclomethasone Dipropionate in solution of 1 – 5mcg/ml concentration & range 515nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Beclomethasone Dipropionate in mcg\ml

Peak area of

1.

1mcg\ml

680

2.

2mcg\ml

742

3.

3mcg\ml

791

4.

4mcg\ml

839

5.

5mcg\ml

872

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

04-16

BECLOSAR-NC CREAM

Prepared By Date of Issue/Effective Date : 08/02/2008

Checked By/ Approved By Review Date : 07/02/2010

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Beclomethasone Dipropionate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Beclomethasone Dipropionate in solution of 1 mcg/ml concentration & range 515nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

1mcg\ml

680

2.

1mcg\ml

682

3.

1mcg\ml

681

4.

1mcg\ml

680

5.

1mcg\ml

685

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

05-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Beclomethasone Dipropionate used = 1mcg\ml. Number of test solution used = 5 680+682+681+680+685 Average area = ----------------------------------5

Dev. S =

= 681.6

(680-681.6)2 +(682-681.6)2 +(681-681.6)2 +(680-681.6)2 +(685-681.6)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 2.56+0.16+0.36+2.56+11.56 = [---------------------------------------]1/2 4 17.2 = [ ----- ----]1/2 4 = [ 4.3 ] 1/2 = 2.0736

RSD

Limit

2.0736 x 100 = -----------------681.6

= 0.3042 %

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

06-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Beclomethasone Dipropionate in Beclosar-NC Cream shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Measure Sample d gm Absorbance at of Ref. Std.

0.0498 0.0492 0.0502

1.9969 2.0013 1.9975

683 685 684

Measured Absorbance at 515nm of sample

% of Beclome thasone

Wt. of Ref. Standard gm.

Wt. of Sample ml

Measured Absorbance at of Ref. Std.

Measured Absorbance at 515nm of sample

% of Beclomet hasone

681 686 686

99.52 98.53 100.88

0.0506 0.0494 0.0497

1.9978 1.9982 2.0018

682 682 684

684 684 686

100.62 99.19 99.61

Label Claim 0.025% w/w Mean 99.64% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 99.89 % Standard Deviation : ±10%

Label Claim Mean Standard Deviation

0.025% w/w 100.14% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

07-16

BECLOSAR-NC CREAM

Prepared By Date of Issue/Effective Date : 08/02/2008

Checked By/ Approved By Review Date : 07/02/2010

Revision No. : 00

METHOD : (Neomycin Sulphate) 1.

Weigh accurately 100mg. of Neomycin Sulphate and dissolve in 100ml. of phosphate buffer pH 8.0 and prepare the dilution 100mcg/ml. in phosphate buffer.

2.

Weight accurately sample (cream) equivalent to 10mg of Neomycin Sulphate, suspend in 25ml. of phosphate buffer and extract with Chloroform (3 x 30ml). Collect chloroform in a dry 100ml volumetric flask filtering each layer through same anhydrous sodium Sulphate bed. Rinse sodium Sulphate layer with chloroform, the combined chloroform layer is made up of volume. Aqueous buffered layer heated in water bath till no perceptible smell of chloroform is diluted to 100ml with phosphate buffer and used for estimation of Neomycin Sulphate.

3.

Pipette 5ml, 6ml, 7ml, 8ml & 9ml of standard and sample solution, add 5ml. of freshly prepared ninhydrin reagent and heat in water bath for 20 minutes, cool, add 2 ml. of methanol and dilute to 50ml with phosphate buffer.

4.

Measure the absorbance without delay at maximum at about 570nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Neomycin Sulphate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Neomycin Sulphate solution serial no. on Xaxis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

08-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.

Linearity of assay method of Neomycin Sulphate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Neomycin Sulphate in solution of 10 – 18mcg/ml concentration & range 570nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Neomycin Sulphate in mcg\ml

Peak area of

1.

10mcg\ml

652

2.

12mcg\ml

698

3.

14mcg\ml

749

4.

16mcg\ml

805

5.

18mcg\ml

862

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

09-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

REPEATIBILITY STUDY

Revision No. : 00

Repeatibility of assay method of Neomycin Sulphate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Neomycin Sulphate in solution of 10mcg/ml concentration & range 570nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

10mcg\ml

650

2.

10mcg\ml

652

3.

10mcg\ml

654

4.

10mcg\ml

648

5.

10mcg\ml

657

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

10-16

BECLOSAR-NC CREAM

Prepared By Date of Issue/Effective Date : 08/02/2008

Checked By/ Approved By Review Date : 07/02/2010

RELATIVE STANDARD DEVIATION

Revision No. : 00

Test concentration of Neomycin Sulphate used = 10mcg\ml. Number of test solution used = 5 650+652+654+648+657 Average area = ----------------------------------5

Dev. S =

= 652.2

(650-652.2)2 +(652-652.2)2 +(654-652.2)2 +(648-652.2)2 +(657-652.2)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 4.84+0.04+3.24+17.64+23.04 = [---------------------------------------]1/2 4 48.8 = [ ----- ----]1/2 4 = [ 12.2 ] 1/2 = 3.493

RSD

Limit

3.493 x 100 = -----------------652.2

= 0.536 %

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

11-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

RUGGEDNESS STUDY

Revision No. : 00

The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Neomycin Sulphate in Beclosar-NC Cream shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm.

Wt. of Measure Sample d gm. Absorbance at of Ref. Std.

0.0998 0.0995 0.1008

2.0256 2.0106 1.9994

656 650 651

Measured Absorbance at 570nm of sample

% of Neomyci n

Wt. of Ref. Standard gm.

Wt. of Sample gm.

Measured Absorbance at of Ref. Std.

Measured Absorbance at 570nm of sample

% of Neomyci n

652 652 649

98.03 99.37 100.62

0.0995 0.0998 0.1029

2.0160 2.0059 1.9989

652 653 656

649 654 658

98.37 99.71 103.33

Label Claim 0.5% w/w Mean 99.34% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 99.89% Standard Deviation : ±10

Label Claim Mean Standard Deviation

0.5% w/w 100.45 % 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

BECLOSAR-NC CREAM

Prepared By Date of Issue/Effective Date : 08/02/2008

METHOD :

12-16

Checked By/ Approved By Review Date : 07/02/2010

Revision No. : 00

(Clotrimazole) 1.

Weigh accurately reference standard 100mg. of Clotrimazole and dissolve in 100ml. of 0.1M hydrochloric acid and prepare the dilution 100mcg./ml of Clotrimazole.

2.

Weigh accurately sample equivalent to 100mg. of Clotrimazole and dissolve in 100ml. of 0.1M hydrochloric acid and prepare the dilution 100mcg./ml of Clotrimazole.

3.

Pipette 1ml, 2ml, 3ml, 4ml & 5ml of standard and sample dilution, add 3ml. of citric acid solution and 2ml of dye solution. Extract the colour with three 8 ml portions of chloroform. Make up the volume to 25ml and pass through anhydrous sodium Sulphate.

4.

Measure the absorbance without delay at maximum at about 422nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Clotrimazole peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Clotrimazole solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

13-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

LINEARITY STUDY

Revision No. : 00

The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Clotrimazole in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Clotrimazole in solution of 4 – 20mcg/ml concentration & range 422nm. Linearity curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Clotrimazole in mcg\ml

Peak area of

1.

4mcg\ml

595

2.

8mcg\ml

632

3.

12mcg\ml

682

4.

16mcg\ml

735

5.

20mcg\ml

795

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

14-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Clotrimazole in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Clotrimazole in solution of 4mcg/ml concentration & range 422nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

4mcg\ml

595

2.

4mcg\ml

592

3.

4mcg\ml

593

4.

4mcg\ml

595

5.

4mcg\ml

596

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

15-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

Revision No. : 00

RELATIVE STANDARD DEVIATION

Test concentration of Neomycin Sulphate used = 4mcg\ml. Number of test solution used = 5 595+592+593+595+596 Average area = ----------------------------------5

Dev. S =

= 594.2

(595-594.2)2 +(592-594.2)2 +(593-594.2)2 +(595-594.2)2 +(596-594.2)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.64 + 4.84 + 1.44 + 0.64 + 3.24 = [------------------------------------------]1/2 4 10.8 = [ ----- ----]1/2 4 = [ 2.7 ] 1/2 = 1.6431

RSD

Limit

1.6431 x 100 = -----------------594.2

= 0.276 %

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/11

16-16

BECLOSAR-NC CREAM

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 08/02/2008

Review Date : 07/02/2010

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Clotrimazole in Beclosar-NC Cream shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Wt. of Measured Ref. Sample gm AbsorStandard bance at gm of Ref. Std.

0.0997 0.1005 0.9989

10.0056 9.9969 10.0125

Measur ed Absorbance at 422nm of sample

% of Clotrima zole

Wt. of Ref. Standard gm

Wt. of Sample gm

Measured Absorbance at of Ref. Std.

Measured Absorbance at 422nm of sample

% of Clotrimaz ole

598 593 590

100.03 100.76 99.32

0.1002 0.0998 0.1012

9.9969 9.9982 10.0015

593 598 597

595 595 599

100.57 99.32 101.53

596 592 593

Label Claim 1% w/w Mean 100.03% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.25% Standard Deviation : ±10%

Label Claim Mean Standard Deviation

1% w/w 100.47% 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation

QC/VLD/12

01-11

Name of the Product:

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Diclofenac Diethylamine & Methylsalicylate in Movsar Gel will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Movsar Gel. SCOPE :

This method is applies to the procedure which is used for calculation and assay of Diclofenac diethylamine & Methylsalicylate.

RESPONSIBILITY: 1. Quality Control Manager. 2. Asstt. Quality Control Chemist.

VALIDATION PARAMETERS: 1. Linearity. 2. Repeatibility. 3. Relative Standard Deviation. 4. Ruggedness.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation

QC/VLD/12

02-11

Name of the Product:

MOVSAR GEL

Prepared By Date of Issue/Effective Date : 04/11/2007

Checked By/ Approved By Review Date : 03/11/2009

Revision No. : 00

METHOD : (Diclofenac diethylamine ) 1.

Weigh accurately 50mg. of Diclofenac sodium working standard in a 100ml volumetric flask. Dissolve with phosphate buffer and make up to 100ml with same solution and prepared the dilution 50mcg./ml. of Diclofenac sodium.

2.

Weight accurately sample equivalent to 5mg of Diclofenac sodium in a 100ml volumetric flask and dissolve in 100ml phosphate buffer.

3.

Pipette 10ml, 12ml, 15ml, 18ml & 20ml of standard and sample solution separately to each funnel add 5ml of methylene blue solution and extract with (10+10+5ml) of chloroform passing each extract through anhydrous sodium Sulphate, dilute to 25ml with chloroform mix to make homogeneous.

4.

Measure the absorbance without delay at maximum at about 650nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Diclofenac diethylamine peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Diclofenac diethylamine solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation

QC/VLD/12

03-11

Name of the Product:

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Diclofenac diethylamine in Movsar Gel shall be studied by measuring the absorbance of the standard Diclofenac diethylamine in solution of 20 – 40mcg/ml concentration & range 650nm. Linearity curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Diclofenac diethylamine in mcg\ml

Peak area of

1.

20mcg\ml

716

2.

24mcg\ml

742

3.

30mcg\ml

795

4.

36mcg\ml

842

5.

40mcg\ml

878

Sarthak Biotech Pvt. Ltd.

Total Page:

04-11

Assay Method Validation Name of the Product:

QC/VLD/12

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Diclofenac diethylamine in Movsar Gel shall be studied by measuring the absorbance of the standard Diclofenac diethylamine in solution of 20 mcg/ml concentration & range 650nm. Repeatibility curve shall be plotted for absorbance against concentration. OBSERVATION TABLE FOR REPEATIBILITY Sr.No.

Concentration in mcg\ml

Peak area

1

20mcg\ml

719

2.

20mcg\ml

714

3.

20mcg\ml

718

4.

20mcg\ml

722

5.

20mcg\ml

720

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/12

05-11

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION

Test concentration of Diclofenac diethylamine used = 20mcg\ml. Number of test solution used = 5 719+714+718+722+720 Average area = ----------------------------------5

Dev. S =

= 718.6

(719-718.6)2 +(714-718.6)2 +(718-718.6)2 +(722-718.6)2 +(720-718.6)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 0.16+21.16+ 0.36+ 11.56+ 1.96 = [------------------------------------------]1/2 4 35.2 = [ ----- ----]1/2 4 = [ 8.8 ] 1/2 = 2.9664

RSD

Limit

2.9664 x 100 = -----------------718.6 :

= 0.412 %

( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/12

06-11

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Diclofenac diethylamine in Movsar Gel shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm

Wt. of Measured Measured Sample Absorba Absorbanc gm. nce at e at of Ref. 650nm of Std. sample

Wt. of Ref. Standard gm.

Wt. of Sample gm.

Measured Absorbance at of Ref. Std.

Measured Absorbance at 650nm of sample

0.0493 0.0501 0.0499

0.5012 0.5006 0.4993

0.0498 0.0494 0.0502

0.4992 0.4984 0.5012

718 716 719

720 718 721

715 718 719

718 716 716

% of

Diclof enac diethyl amine 99.78 99.80 99.52

Label Claim 1.16% w/w Mean 99.36% Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 99.88 % Standard Deviation : ±10%

Label Claim Mean Standard Deviation

100.03 99.39 100.44

1.16% w/w 100.44 10%

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/12

MOVSAR GEL

% of

Diclofe nac diethyl amine

07-11

Prepared By Date of Issue/Effective Date : 04/11/2007

Checked By/ Approved By Review Date : 03/11/2009

Revision No. : 00

METHOD : (Methylsalicylate) 1.

Weigh accurately 100mg. of Methylsalicylate as working standard in a 100ml volumetric flask and dissolve in 0.02M hydrochloric acid and make up to produced 100ml, again dilute 15ml of the resulting solution to 100ml with 0.02M hydrochloric acid.

2.

Weight accurately equivalent to 100mg of Methyl salicylate in a 500ml conical flask add 30ml of 0.5M sodium hydroxide and 20ml of methyl alcohol, mix and reflux for one hour on steam bath to sponify and hydrolyse, cool to room temperature and add 20ml of 1.0M hydrochloric acid and dilute to 100ml with water and filter. Further dilute 15ml to 100ml with 0.02M hydrochloric acid.

3.

Pipette 10ml, 15ml, 20ml, 25ml & 30ml of standard and sample solution in separately funnel and extract with 25ml of chloroform, separate chloroform layer in a dry test tube. To 5 ml of chloroform layer add 10ml of ferric ammonium sulfate reagent and separate the aqueous layer.

4.

Measure the absorbance without delay at maximum at about 530nm, against respective blank and tabulate the observed data in the observation table.

5.

Plot linearity curve from the different concentration corresponding area of Methylsalicylate peak. Taking concentration on X-axis and corresponding peak on Yaxis. Plot graph should be connected at least three points.

6.

Plot repeatability curve from by taking Methylsalicylate solution serial no. on X-axis and observed peak area on Y-axis.

7.

Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .

8.

Calculated the result by comparison with peak area obtained in standard and sample solution respectively.

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation

QC/VLD/12

08-11

Name of the Product:

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

LINEARITY STUDY The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range. Linearity of assay method of Methylsalicylate in Movsar Gel shall be studied by measuring the absorbance of the standard Methylsalicylate in solution of 30 – 90mcg/ml concentration & range 530nm. Linearity curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR LINEARITY Sr.No. Concentration of Methylsalicylate in mcg\ml

Peak area

1.

30mcg\ml

685

2.

45mcg\ml

752

3.

60mcg\ml

796

4.

75mcg\ml

855

5.

90mcg\ml

888

Sarthak Biotech Pvt. Ltd.

Total Page:

Assay Method Validation Name of the Product:

QC/VLD/12

09-11

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

REPEATIBILITY STUDY Repeatibility of assay method of Methylsalicylate in Movsar Gel shall be studied by measuring the absorbance of the standard Methylsalicylate in solution of 30mcg/ml concentration & range 530nm. Repeatibility curve shall be plotted for absorbance against concentration.

OBSERVATION TABLE FOR REPEATIBILITY

Sr.No.

Concentration in mcg\ml

Peak area

1

30mcg\ml

687

2.

30mcg\ml

685

3.

30mcg\ml

684

4.

30mcg\ml

686

5.

30mcg\ml

685

Sarthak Biotech Pvt. Ltd.

Total Page:

10-11

QC/VLD/12

Assay Method Validation Name of the Product:

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

RELATIVE STANDARD DEVIATION

Test concentration of Methylsalicylate used = 30mcg\ml. Number of test solution used = 5 687+685+684+686+685 Average area = ----------------------------------5

Dev. S =

= 685.4

(687-685.4)2 +(685-685.4)2 +(684-685.4)2 +(686-685.4)2 +(685-685.4)2 [ ---------------------------------------------------------------------------------------] 1/2 5-1 2.56+0.16+1.96+0.36+0.16 = [----------------------------------------]1/2 4

RSD

Limit

=

5.2 [ ----- ----]1/2 4

=

[ 1.3 ]

=

1.1401

1.1401 x 100 = -----------------685.4

1/2

= 0.166 %

: ( Not more than 2.0% )

Sarthak Biotech Pvt. Ltd.

Total Page:

11-11

Assay Method Validation Name of the Product:

QC/VLD/12

MOVSAR GEL

Prepared By

Checked By/ Approved By

Date of Issue/Effective Date : 04/11/2007

Review Date : 03/11/2009

Revision No. : 00

RUGGEDNESS STUDY The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test conditions (i.e. different laboratories, analysts, days & Instruments). The ruggedness of assay method of Methylsalicylate in Movsar Gel shall be studied by analyzing the samples from same lot by two different analysts from same laboratory or at two different laboratories by two different analysts on two different days. RUGGEDNESS STUDY DATA Ruggedness data for assay method by two different analysts on different days and different instrument. Sr. No.

01 02 03

Analyst- I (Satpal Saini) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Analyst- II (Rajnish Kohli) Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117 Location : Instrument Lab.

Wt. of Ref. Standard gm

Wt. of Measure Measured Sample d Absorbance gm Absorb at 530nm ance at of sample of Ref. Std.

% of Methylsa licylate

Wt. of Ref. Standard gm

Wt. of Sample gm.

Measured Absorbanc e at of Ref. Std.

Measured Absorban ce at 530nm of sample

% of Methylsal icylate

0.0998 0.1009 0.0998

0.9994 0.9989 1.0012

100.15 100.87 99.39

0.1018 0.0998 0.1009

0.9996 1.0015 0.9994

687 684 684

686 686 682

101.68 99.93 100.66

684 684 687

686 685 685

Label Claim 10% w/w Mean 100.31 % Standard Deviation 10% Combining mean of I & II of both the analysts : Mean : 100.53% Standard Deviation : ±10%

Label Claim Mean Standard Deviation

10% w/w 100.75% 10%