Sarthak Biotech Pvt. Ltd.
Total Page: 01-06
Assay Method Validation
QC/VLD/01Name of the Product:
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
OBJECTIVE : The establish documented evidence which will provide a high degree of
assurance that the analytical method for assay of Ciprofloxacin in Sarcip-500 Tablet will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarcip-500 Tablet.
SCOPE : 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: 02-06
Assay Method Validation
QC/VLD/01Name of the Product:
SARCIP-500 TABLET
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009 Revision No. : 00
METHOD :
1. Weight accurately equivalent to about 100mg of Ciprofloxacin working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with 0.1N- Hydrochloric 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: 03-06
Assay Method Validation
QC/VLD/01Name of the Product:
SARCIP-500 TABLET
Prepared By 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
Sarthak Biotech Pvt. Ltd.
Total Page: 04-06
Assay Method Validation
QC/VLD/01Name of the Product:
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
Assay Method Validation
QC/VLD/01Name of the Product:
SARCIP-500 TABLET
Prepared By 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 = --- = 604.8 5 (604-604.8)2 +(608-604.8)2 +(603-604.8)2 +(605-604.8)2 +(604-604.8)2 Dev. S = [ ---] 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 1.9235 x 100 RSD = --- = 0.318% 604.8
Sarthak Biotech Pvt. Ltd.
Total Page: 06-06
Assay Method Validation
QC/VLD/01Name of the Product:
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. 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. Measured Absor-bance at of Ref. Std. Measured Absor-bance at 438nm of sample % of Cipro-floxacin Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absor-bance at of Ref. Std. Measured Absor-bance at 438nm of sample % of Cipro-floxacin 01 0.1168 0.1348 602 608 100.11 0.1162 0.1349 606 601 99.43 02 0.1156 0.1352 607 612 99.61 0.1165 0.1356 609 616 101.15 03 0.1158 0.1342 605 615 101.35 0.1164 0.1352 612 608 99.55
Average wt of tablet 670.2 mg Averagewt.of tablet 675.1 mg
Mean 100.35% Mean 100.04%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.2 %
Sarthak Biotech Pvt. Ltd.
Total Page: 01-06
Assay Method Validation
QC/VLD/02Name of the Product:
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: 02-06
Assay Method Validation
QC/VLD/02Name of the Product:
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
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 0.1N- Hydrochloric 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: 03-06
Assay Method Validation
QC/VLD/02Name of the Product:
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
Sarthak Biotech Pvt. Ltd.
Total Page: 04-06
Assay Method Validation
QC/VLD/02Name of the Product:
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
Assay Method Validation
QC/VLD/02Name of the Product:
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 = --- = 513.4 5 (515-513.4)2 +(512-513.4)2 +(515-513.4)2 +(509-513.4)2 +(519-513.4)2 1/2 Dev. S = [ ---] 5-1 2.56+1.96+1.96+19.36+31.36 = [---]1/2 4 57.2 = [ ---]1/2 4 = [ 14.3] 1/2 = 3.781 3.781 x 100 RSD = --- = 0.736% 513.4
Limit : ( Not more than 2.0% )
Assay Method Validation
QC/VLD/02Name of the Product:
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.
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. Measured Absorbanc e at of Ref. Std. Measured Absorbance at 437nm of sample % of Ofloxacin Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absorbanc e at of Ref. Std. Measured Absor-bance at 437nm of sample % of Ofloxacin 01 0.0498 0.1021 509 512 100.67 0.0499 0.1006 512 505 98.97 02 0.0501 0.1019 511 515 101.7 0.0512 0.1018 504 510 102.95 03 0.0512 0.1029 519 512 100.7 0.0489 0.1012 508 514 98.90
Average wt of tablet 410.4 mg Averagewt.of tablet 404.6 mg
Mean 101.02% Mean 100.27%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.65 %
Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd.
Total Page: 01-11
Assay Method Validation
QC/VLD/03Prepared 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.
Total Page: 02-11
Assay Method Validation
QC/VLD/03Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
PROCEDURE : (For Paracetamol)
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.
Total Page: 03-11
Assay Method Validation
QC/VLD/03Prepared 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: 04-11
Assay Method Validation
QC/VLD/03Prepared 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.
Total Page: 05-11
Assay Method Validation
QC/VLD/03Prepared 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 = --- = 526.2 5 (525-526.2)2 +(528-526.2)2 +(527-526.2)2 +(526-526.2)2 +(525-526-2)2 1/2 Dev. S = [ ---] 5-1 1.44+3.24 +0.64+0.04+1.44 = [---]1/2 4 6.8 = [ ---]1/2 4 = [ 1.7] 1/2 = 1.303 1.303 x 100 RSD = --- = 0.2477% 526.2
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 06-11
Assay Method Validation
QC/VLD/03Prepared 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.
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. Measured Absor-bance at of Ref. Std. Measured Absor-bance at 450nm of sample % of Paraceta mol-mol Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absor-bance at of Ref. Std. Measured Absor-bance at 450nm of sample % of Paraceta-mol 01 0.0598 0.0784 521 528 100.83 0.0602 0.0782 512 518 101.99 02 0.0601 0.0792 532 528 98.23 0.0592 0.0779 519 515 98.75 03 0.0592 0.0784 518 525 99.19 0.0612 0.0791 522 525 101.9
Average wt of tablet 652.2 mg Averagewt.of tablet 654.8 mg
Mean 99.41% Mean 100.88%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.15 %
Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd.
Total Page: 07-11
Assay Method Validation
QC/VLD/03Name of the Product:
SARNIM-P TABLET
Prepared By Checked By/ Approved By
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: 08-11
Assay Method Validation
QC/VLD/03Name of the Product:
SARNIM-P TABLET
Prepared By Checked By/ Approved By
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: 09-11
Assay Method Validation
QC/VLD/03Name of the Product:
SARNIM-P TABLET
Prepared By Checked By/ Approved By
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.
Total Page: 10-11
Assay Method Validation
QC/VLD/03Name of the Product:
SARNIM-P TABLET
Prepared By Checked By/ Approved By
RELATIVE STANDARD DEVIATION
Test concentration of Nimesulide used = 20mcg\ml.
Number of test solution used = 5 559 + 561 + 561 + 560 + 563 Average area = --- = 560.8 5 (559-560.8)2 +(561-560.8)2 +(561-560.8)2 +(560-560.8)2 +(563-560.8)2 1/2 Dev. S = [ ---] 5-1 3.24+0.04 +0.04+0.64+4.84 = [---]1/2 4 8.8 = [ ---]1/2 4 = [ 2.2] 1/2 = 1.4142 1.4142 x 100 RSD = --- = 0.2521% 560.8
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 11-11
Assay Method Validation
QC/VLD/03Name of the Product:
SARNIM-P TABLET
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.
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. Measured Absor-bance at of Ref. Std. Measured Absor-bance at 405nm of sample % of Nimesuli de Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absor-bance at of Ref. Std. Measured Absor-bance at 405nm of sample % of Nimesuli de 01 0.0499 0.3258 560 568 101.31 0.0498 0.3261 565 560 99.11 02 0.0489 0.3251 542 545 98.64 0.0496 0.3269 559 562 99.88 03 0.0501 0.3251 560 555 99.61 0.0502 0.3251 553 556 101.65
Average wt of tablet 652.2 mg Averagewt.of tablet 654.8 mg
Mean 99.85% Mean 100.21%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.03 %
Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd.
Total Page: 01-06
Assay Method Validation
QC/VLD/04Name of the Product:
SARFIX-100 DT TABLET
Prepared By Checked By/ Approved By
PURPOSE :- The establish documented evidence which will provide a high degree of
assurance that the analytical method for assay of Cefixime Trihydrate in Sarfix-100 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.
Sarthak Biotech Pvt. Ltd.
Total Page: 02-06
Assay Method Validation
QC/VLD/04Name of the Product:
SARFIX-100 DT TABLET
Prepared By Checked By/ Approved By
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: 03-06
Assay Method Validation
QC/VLD/04Name of the Product:
SARFIX-100 DT TABLET
Prepared By Checked By/ Approved By
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: 04-06
Assay Method Validation
QC/VLD/04Name of the Product:
SARFIX-100 DT TABLET
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 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.
Total Page: 05-06
Assay Method Validation
QC/VLD/04Name of the Product:
SARFIX-100 DT TABLET
Prepared By Checked By/ Approved By
RELATIVE STANDARD DEVIATION
Test concentration of Cefixime used = 10mcg\ml.
Number of test solution used = 5 495 + 492 + 497 + 495 + 496 Average area = --- = 495 5 (495-495)2 +(495-492)2 +(495-497)2 +(495-495)2 +(495-496)2 1/2 Dev. S = [ ---] 5-1 0+9+4+0+1 = [---]1/2 4 14 = [ ---]1/2 4 = [ 3.5] 1/2 = 1.8708 1.8708 x 100 RSD = --- = 0.377% 495
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 06-06
Assay Method Validation
QC/VLD/04Name of the Product:
SARFIX-100 DT TABLET
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 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. 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. Measured Absor-bance at of Ref. Std. Measured Absor-bance at 268nm of sample % of Cefixime Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absor-bance at of Ref. Std. Measured Absor-bance at 268nm of sample % of Cefixime 01 0.1128 0.4019 498 502 101.75 0.1118 0.4028 502 508 101.21 02 0.1119 0.4021 492 499 101.50 0.1129 0.4031 499 505 102.14 03 0.1131 0.4012 498 488 99.34 0.1125 0.4022 492 498 102.02
Average wt of tablet 402.8 mg Averagewt.of tablet 403.6 mg
Mean 100.86% Mean 101.79%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 101.32 %
Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd.
Total Page: 01-06
Assay Method Validation
QC/VLD/05Name of the Product:
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
PURPOSE: 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: 02-06
Assay Method Validation
QC/VLD/05Name of the Product:
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
METHOD :
1. 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: 03-06
Assay Method Validation
QC/VLD/05Name of the Product:
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: 04-06
Assay Method Validation
QC/VLD/05Name of the Product:
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
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. Concentration in mcg\ml Peak area
1 40mcg\ml 651 2 . 40mcg\ml 653 3. 40mcg\ml 652 4. 40mcg\ml 657 5. 40mcg\ml 651
Sarthak Biotech Pvt. Ltd.
Total Page: 05-06
Assay Method Validation
QC/VLD/05Name of the Product:
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
RELATIVE STANDARD DEVIATION
Test concentration of Amoxycillin used = 40mcg\ml.
Number of test solution used = 5 651+653+652+657+651 Average area = --- = 652.8 5 (651-652.8)2 +(653-652.8)2 +(652-652.8)2 +(657-652.8)2 +(651-652.8)2 Dev. S = [ ---] 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 2.489 x 100 RSD = --- = 0.381% 652.8
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 06-06
Assay Method Validation
QC/VLD/05Name of the Product:
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
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.
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. Measured Absor-bance at of Ref. Std. Measured Absor-bance at 605nm of sample % of Amoxyci -llin Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absor-bance at of Ref. Std. Measured Absor-bance at 605nm of sample % of Amoxycil -lin 01 0.1162 0.1148 655 652 100.79 0.1154 0.1168 659 655 98.57 02 0.1159 0.1162 654 658 100.38 0.1162 0.1169 652 662 101.30 03 0.1165 0.1155 653 657 101.51 0.1152 0.1162 656 650 98.60
Average fill weight 580.2 mg Average fill weight 582.2 mg
Mean 100.89% Mean 99.49%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.19 %
Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd.
Total Page: 01-06
Assay Method Validation
QC/VLD/06Name of the Product:
ACIGONE CAPSULE
Prepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
PURPOSE: 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: 02-06
Assay Method Validation
QC/VLD/06Name of the Product:
ACIGONE CAPSULE
Prepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
METHOD :
1. 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: 03-06
Assay Method Validation
QC/VLD/06Name of the Product:
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: 04-06
Assay Method Validation
QC/VLD/06Name of the Product:
ACIGONE CAPSULE
Prepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 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. Concentration in mcg\ml Peak area
1 8mcg\ml 398 2 . 8mcg\ml 401 3. 8mcg\ml 397 4. 8mcg\ml 399 5. 8mcg\ml 398
Sarthak Biotech Pvt. Ltd.
Total Page: 05-06
Assay Method Validation
QC/VLD/06Name of the Product:
ACIGONE CAPSULE
Prepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Amoxycillin used = 8mcg\ml.
Number of test solution used = 5 398+401+397+399+398 Average area = --- = 398.6 5 (398-398.6)2 +(401-398.6)2 +(397-398.6)2 +(399-398.6)2 +(398-398.6)2 Dev. S = [ ---] 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 1.5165 x 100 RSD = --- = 0.3804% 398.6
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 06-06
Assay Method Validation
QC/VLD/06Name of the Product:
ACIGONE CAPSULE
Prepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 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.
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. Measured Absor-bance at of Ref. Std. Measured Absor-bance at 268nm of sample % of Omepraz -ole Wt. of Ref. Standard gm. Wt. of Sample gm Measured Absor-bance at of Ref. Std. Measured Absor-bance at 268nm of sample % of Omepra-zole 01 0.2657 0.2940 395 399 100.40 0.2672 0.2926 403 399 99.48 02 0.2673 0.2927 401 396 99.20 0.2657 0.2961 398 401 99.47 03 0.2661 0.2939 393 396 100.38 0.2673 0.2926 396 393 99.75
Average fill weight 293.4 mg Average fill weight 293.4 mg
Mean 99.99% Mean 99.57%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.78
Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd.
Total Page: 01-06
Assay Method Validation
QC/VLD/07Name of the Product:
SARMOX DRY SYRUP
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
PURPOSE: 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: 02-06
Assay Method Validation
QC/VLD/07Name of the Product:
SARMOX DRY SYRUP
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
METHOD :
1. 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: 03-06
Assay Method Validation
QC/VLD/07Name of the Product:
SARMOX DRY SYRUP
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 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: 04-06
Assay Method Validation
QC/VLD/07Name of the Product:
SARMOX DRY SYRUP
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 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. Concentration in mcg\ml Peak area
1 40mcg\ml 651 2 . 40mcg\ml 652 3. 40mcg\ml 657 4. 40mcg\ml 660 5. 40mcg\ml 655
Sarthak Biotech Pvt. Ltd.
Total Page: 05-06
Assay Method Validation
QC/VLD/07Name of the Product:
SARMOX DRY SYRUP
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
Test concentration of Amoxycillin used = 40mcg\ml.
Number of test solution used = 5 651+652+657+660+655 Average area = --- = 655 5 (651-655)2 +(652-655)2 +(657-655)2 +(660-655)2 +(655-655)2 Dev. S = [ ---] 1/2 5-1 16+ 9+ 4 + 25 + 0 = [---]1/2 4 54 = [ ---]1/2 4 = [ 13.5 ] 1/2 = 3.6742 3.6742 x 100 RSD = --- = 0.560% 655
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 06-06
Assay Method Validation
QC/VLD/07Name of the Product:
SARMOX DRY SYRUP
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 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 Sample ml Measure d Absor-bance at of Ref. Std. Measured Absor-bance at 605nm of sample % of Amoxy-cillin Wt. of Ref. Standard gm. Wt. of Sample ml Measured Absor-bance at of Ref. Std. Measured Absor-bance at 605nm of sample % of Amoxy-cillin 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: 01-06
Assay Method Validation
QC/VLD/08Name of the Product:
CEFSAR-125 DRY SYRUP
Prepared By Checked By/ Approved By
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: 02-06
Assay Method Validation
QC/VLD/08Name of the Product:
CEFSAR-125 DRY SYRUP
Prepared By Checked By/ Approved By
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: 03-06
Assay Method Validation
QC/VLD/08Name of the Product:
CEFSAR-125 DRY SYRUP
Prepared By Checked By/ Approved By
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 250mg-to 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: 04-06
Assay Method Validation
QC/VLD/08Name of the Product:
CEFSAR-125 DRY SYRUP
Prepared By Checked By/ Approved By
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: 05-06
Assay Method Validation
QC/VLD/08Name of the Product:
CEFSAR-125 DRY SYRUP
Prepared By Checked By/ Approved By
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 = --- = 7.02 5 (7- 7.02)2 +(6.9-7.02)2 +(7.1-7.02)2 +(7.2-7.02)2 +(6.9-7.02)2 Dev. S = [ ---] 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 0.130 x 100 RSD = --- = 1.857% 7.02
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd.
Total Page: 06-06
Assay Method Validation
QC/VLD/08Name of the Product:
CEFSAR-125 DRY SYRUP
Prepared By Checked By/ Approved By
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.
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 01 10ml 0.102305 7.0ml 99.51 10ml 0.102305 7.1ml 100.94 02 10ml 0.102305 6.9ml 98.09 10ml 0.102305 7.0ml 99.51 03 10ml 0.102305 7.0ml 99.51 10ml 0.102305 7.0ml 99.52
Label Claim 125mg Cephalexin/5ml Label Claim 125mg Cephalexin/5ml
Mean 99.04% Mean 99.99%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.52%
Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd.
Total Page: 01-05
Assay Method Validation
QC/VLD/09Name of the Product:
CHLORTEX MOUTH WASH
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of
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: 02-05
Assay Method Validation
QC/VLD/09Name of the Product:
CHLORTEX MOUTH WASH
Prepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
