analytical method validation report of saxagliptin tablet

Analytical Method Validation ReportSaglip TabletDocument No. CCL-AMVR-170-A

2015

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ANALYTICAL METHOD VALIDATION REPORT Saglip Tablets

DOCUMENT NO.: CCL-AMVR-170-A


Table of Contents

1 INTRODUCTION........................................................................................................................................................ 3

2 ANALYTICAL PROCEDURE......................................................................................................................................... 3

2.1 ASSAY PROCEDURE.........................................................................................................................................................3

3 VALIDATION OF METHOD......................................................................................................................................... 3

3.1 SPECIFICITY...................................................................................................................................................................43.2 LINEARITY....................................................................................................................................................................53.3 RANGE........................................................................................................................................................................73.4 ACCURACY...................................................................................................................................................................83.5 PRECISION..................................................................................................................................................................10

3.5.1 Repeatability......................................................................................................................................................113.5.2 Intermediate precision........................................................................................................................................14

3.6 LIMIT OF DETECTION (LOD) AND LIMIT OF QUANTITATION (LOQ).......................................................................................173.7 ROBUSTNESS..............................................................................................................................................................173.8 SYSTEM SUITABILITY.....................................................................................................................................................233.9 CONCLUSION..............................................................................................................................................................233.10 DEVIATION.................................................................................................................................................................23

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1 IntroductionThe analytical method for Saglip Tablet was validated and found to be valid for assay and stability studies due to its accuracy and specificity. Forced degradation studies of finished pharmaceutical product (Saglip tablet) were performed, the resolution between degraded products and principle peak was found to be >2. Peak purity and relative retention times confirmed the specificity of method. Method was investigated for its range, linearity, accuracy, precision, robustness, and LOQ, LOD and solution stability. All the parameters were performed on Agilent 1260 series HPLC equipped with Auto sampler, column oven, quaternary low pressure gradient pump and PDA detector for peak purity purposes

2 Analytical Procedure2.1 Assay procedureBuffer: Dissolve 2.72 grams of Potassium Dihydrogen Phosphate in 1000 mL of distilled water and adjust the pH 4.6 with dilute Sodium Hydroxide solution.Diluent: 0.1N HClMobile phase: Buffer: Acetonitrile (80:20)Standard solution: Weigh on analytical balance, working standard of Saxagliptin HCl equivalent to 25mg of Saxagliptin and transfer it into a 50ml volumetric flask, make up the volume with diluent and sonicate for 10 minutes. Take 10ml from this solution into a 100ml volumetric flask and make up the volume with diluent.Sample solution: Weigh on analytical balance, ground powder of tablets equivalent to 5mg of Saxagliptin and transfer it into a 100ml volumetric flask, make up the volume with diluent and sonicate for 10 minutes (50ppm). Mode: LCDetector: UV 213 nmColumn: C18Column temperature: AmbientFlow rate: 1.5 mL/min Injection size: 10 µL System suitability requirementsTailing factor: NMT 2.0 Relative standard deviation: NMT 2.0%

Calculate the percentage of the labeled amount of Saxagliptin in the portion of Tablets taken:Result = (rU/rS) x (CS/CU) x 100/1.116

rU = peak response from the Sample solution rS = peak response from the Standard solutionCS = concentration of Saxagliptin HCl RS in the Standard solution (mg/mL)CU = nominal concentration of Saxagliptin in the Sample solution (mg/mL)

Acceptance criteria: 90.0%–110.0%

3 Validation of Method

Following Validation characteristics were considered for analytical method validation:

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1) Specificity (Identification)2) Linearity 3) Range4) Accuracy (Recovery)5) Precision

i. Repeatability ii. Intermediate Precision

6) LOD and LOQ7) Robustness8) Solution stability 9) System suitability

3.1 SpecificitySamples of drug product were exposed to heat, light, acid, base, and oxidizing agent to produce 10%–30% degradation of the active. The degraded samples were then analyzed using the method to determine if there were interferences with the active or related compound peaks.

The following degradation conditions were selected:Obtain or prepare solutions of ~0.1 N HCL, 1 N HCL, 2 N HCL ~0.15 N NaOH, 1 N NaOH, 2 N NaOH ~1% H2O2, 5% H2O2, 10% H2O2 and purified water.

Table 1FORCED DEGRADATION FOR SAXAGLIPTIN

Stress Condition

weight taken (mg) Chrom. No.

Peak Area

Mean Area

No. of degradation product

(Area; RT)

Theoretical Content

(µg mL−1)

Amount recovered (µg mL−1)

Recovery (%)

Degradation

Unstressed sample

231.011 606.7

607.2 0 2.49 2.76 99.52% 0.48%2 608.53 606.5

0.1 N HCl 226.054 666.7

668.4 0 2.43 3.04 111.95% -11.95%5 667.46 671.1

1 N HCl 229.947 669.5

669.1 0 2.48 3.05 110.17% -10.17%8 668.39 669.4

2 N HCl 229.8210 675.8

675.8 0 2.48 3.08 111.33% -11.33%11 673.812 677.7

0.1N NaOH 228.7113 443.6

443.9(4)

2.2,3.6,3.8,7.12.46 2.02 73.49% 26.51%14 443.6

15 444.6

1 N NaOH 233.5516 40.8

40.8(6)

2.2,3.6,3.8,7.1,1.7,4.52.52 0.19 6.61% 93.39%17 41.1

18 40.4

2 N NaOH 227.219 0.0

0.0(6)

2.2,3.6,3.8,7.1,1.7,4.52.45 0.00 0.00% 100.00%20 0.0

21 0.01 %

H2O2229.99 22 664.5 665.0 (1)

3.82.48 3.03 109.47% -9.47%

23 665.4

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Table 1FORCED DEGRADATION FOR SAXAGLIPTIN

24 665.0

5 %H2O2

231.2825 667.3

667.1(2)

3.8,3.62.49 3.04 109.20% -9.20%26 667.1

27 666.8

10 %H2O2

232.3728 601.0

601.3 0 2.50 2.74 97.97% 2.03%29 602.830 600.0

Stan

dard

25.12

34 555.4

551.9

-----

35 551.036 550.237 553.038 550.739 551.2

Acceptance Criteria: Peak purity should be NLT 990 Resolution should be more than 2.0

Conclusion:Resolution was found to be more than 2.0 (See chromatograms)Peak purity was found to be more than 990 (See chromatograms)Relative retention times are mentioned in table and are different for degraded products from active.3.2 LinearityA stock solution of Saxagliptan (0.5mg mL−1 saxagliptan) was prepared by dissolving 1000 mg drug in 100 mL Diluent. Solutions of different concentration (30--70 µg mL−1) for construction of calibration plots were prepared from this stock solution. The mobile phase was filtered through a 0.22-µm membrane filter and delivered at 1.50 mL min−1 for column equilibration; the baseline was monitored continuously during this process. The detection wavelength was 213 nm. The prepared dilutions were injected at the rate of 10µl in series, peak area was calculated for each dilution, and concentration was plotted against peak area.

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Table 2

Linearity study of Saxagliptin Sr. # Chrom. # Conc. (ppm) Peak Area MEAN RSD (%)

1 4 30 222.3223.2 0.339%2 5 30 223.5

3 6 30 223.74 7 40 299.4

298.4 0.355%5 8 40 298.66 9 40 297.37 10 50 372.1

372.8 0.217%8 11 50 372.79 12 50 373.7

10 13 60 447.2447.1 0.169%11 14 60 446.3

12 15 60 447.813 16 70 518.7

519.1 0.077%14 17 70 519.515 18 70 519.1

25 30 35 40 45 50 55 60 65 70 750

100

200

300

400

500

600

f(x) = 7.40533333333334 x + 1.85999999999996R² = 0.999898962830931

Linear plot for Saglip Tablet

Concentration

Peak

Are

a

Acceptance Criteria: Coefficient of determination (r2) should be greater than 0.990. The y intercept should not significantly depart from zero Area response of y intercept should be less than 5% of the response of the midrange concentration

value.Conclusion: of 24


The calibration plot of peak area against concentration was linear in the range investigated (30–70 µg mL−1). Calibration data, with their relative standard deviations, % RSD and standard error are listed in Table I. The low values of RSD and standard error show the method is precise. The linear regression data for the calibration plot are indicative of a good linear relationship between peak area and concentration over a wide range. The linear regression equation was y =1468.5x + 40069 and the re-gression coefficient was 0.9999.3.3 Range A stock solution of Saxagliptan (0.5mg mL−1 saxagliptan) was prepared by dissolving 1000 mg drug in 100 mL Diluent. Solutions of different concentration (20--100 µg mL−1) for construction of calibration plots were prepared from this stock solution. The mobile phase was filtered through a 0.22-µm membrane filter and delivered at 1.50 mL min−1 for column equilibration; the baseline was monitored continuously during this process. The detection wavelength was 213 nm. The prepared dilutions were injected at the rate of 10µl in series, peak area was calculated for each dilution, and concentration was plotted against peak area.

Table 3Range study of Sitagliptin

Sr. # Conc. (ppm) Peak Area MEAN RSD (%)

1 20 147.4148 0.434%2 20 148.4

3 20 148.64 30 222.3

223 0.339%5 30 223.56 30 223.77 40 299.4

298 0.355%8 40 298.69 40 297.3

10 50 372.1373 0.217%11 50 372.7

12 50 373.713 60 447.2

447 0.169%14 60 446.3

15 60 447.8

16 70 518.7519 0.077%17 70 519.5

18 70 519.119 80 600.3

600 0.025%20 80 600.421 80 600.1

22 100 741.6742 0.075%23 100 742.7

24 100 742.0

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10 20 30 40 50 60 70 80 90 100 1100

100

200

300

400

500

600

700

800

f(x) = 7.44456140350877 x + 0.135087719298383R² = 0.999870806227799

Concentration (ppm) Line Fit Plot for Range study of Saglip Tablet

Concentration (ppm)

Peak

Are

a

ConclusionThe calibration plot of peak area against concentration was linear in the range investigated (20-100 µg mL−1). Calibration data, with their relative standard deviations, % RSD and standard error are listed in Table I. The low values of RSD and standard error show the method is precise. The linear regression data for the calibration plot are indicative of a good linear relationship between peak area and concentration over a wide range. The linear regression equation was y = 1412.1x + 13700 and the regression coefficient was 0.9999.Acceptance CriteriaRegression Coefficient > 0.990

3.4 Accuracy Standard preparation:Weight accurately about 50 mg of Saxagliptin in a 100 ml volumetric flask and dissolve it in 0.1N HCl and make up volume 0.1N HCl. Take 10ml from this dilution in 100ml volumetric flask and makeup volume with 0.1N HCl.Sample preparation:Weight accurately about 195.0 mg of placebo in a 100 ml volumetric flask (thre different flasks) and dissolve it in water. (A, B, C) 2-A stock solution of Saxagliptan HCl (0.5mg mL−1) was prepared by dissolving 50 mg of Saxagliptan HCl in 100 mL 0.1N HCl. Transfer 8, 10, and 12 ml of this solution in three different 100 ml volumetric flasks A, B, C respectively containing placebo. Make up volume with diluent to get three concentration levels of 80%, 100% and 120% respectively.

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Accuracy Set 1Potency of WS: 99.10% Prop Wt: 230mgWeight of placebo: 195.00 Weight of active: 50.04

Table 4Accuracy SET 1

Weight of placebo

mg

Volume ofstock

solution(ml)

Theoretical Content (µg/ml) Peak Area Mean Area RSD

Actual Recovery

Content (µg/ml)%age

140.30 8.039.67

431.9434 0.49 39.34 99.18434.3

436.1

140.28 10.049.59

541.9539 0.40 48.89 98.59537.8

538.6

140.26 12.059.51

656.5656 0.07 59.46 99.93655.6

656.1

Working Standard 50.04 -----

551.1

552.10 0.26

Mean recovery (%) 99.23552.6

550.9

554.4

551.5

----

Accuracy Set 2Potency of WS: 99.10% Prop Wt: 150mgWeight of placebo: 140.00 Weight of active: 50.23


Weight of placebo

mg

Volume ofstock solution

(ml)

Theoretical Content (µg/ml) Peak Area Mean Area RSD

Actual Recovery

Content (µg/ml)%age

140.30 8.039.74

438438 0.15 39.70 99.91438.7

437.4

140.28 10.049.59

545.2544 0.14 49.34 99.49543.9

543.9

140.26 12.059.73

652.0652 0.02 59.11 98.95652.3

652.1Working Standard 50.23 551.1 552.10 0.26


550.9

554.4

551.5

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-----

Accuracy Set 3Potency of WS: 99.10% Prop Wt: 150 mgWeight of placebo: 140.00 Weight of active: 49.79


Weight of placebo

mg

Volume of Active

mlTheoretical Content (%) Peak Area Mean Area RSD

Actual Recovery

Content (w/w %)%age

140.01 8.0

39.39

437.1

437 0.08 39.74 100.88436.8

436.4

140.54 10.0

49.59

531.4

532 0.17 48.41 97.62533.1

531.8

140.07 12.0

59.51

644.6

645 0.38 58.71 98.66643.3

648

Working Standard 49.79

542.8

543.14 0.09


543.2

543.2

543.9

The recovery of the method is determined by spiking a placebo with active pharmaceutical ingredient of known potency. The values of recovery (%), RSD (%), and SEM listed in Table 4, 5 and 6 listed above indicate that the method is accurate.Acceptance Criteria:

The percent recovery of the spiked placebos should be within 100±2.0% for the average of each set of three weights.

Each individual sample recovery should lie within the range of 98% to 102%.

Conclusion:Hence above acceptance criteria for accuracy is met, therefore method is accurate3.5 Precision Precision was determined as both repeatability and intermediate precision. For repeatability six replicates of a sample were analyzed on three different days and their RSD was calculated. For intermediate precision same

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sample was analyzed on two different equipments, columns, days and by two different analyst and their standard deviation and relative standard deviations were recorded

3.5.1 Repeatability

Tablet 7Set 1

Potency of standard 99.10% % Weight of standard taken 51.4 mgWeight of sample taken 205.46 mg Proposed weight / Tablet 210 mgFactor 1.116

Standard Peak Areas Sample Peak AreasSr # Area

RSD(%)

Sr # Area1 365.6 1 274.12 361.6 2 274.43 362.6 3 274.44 363.3

0.39

Average274.30

Result5 362.66 362.1

RSD (%)0.06 70.51%Average 363.0

Tablet 8Set 2



RSD(%)

Sr # Area1 349.6 1 301.52 350.2 2 302.33 348.5 3 302.44 348.9

0.33

Average302.07

Result5 346.86 348.5

RSD (%)0.16 74.40%Average 348.8

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Tablet 9Set 3



RSD(%)

Sr # Area1 376.2 1 319.32 377.0 2 320.23 376.7 3 316.94 376.7

0.27

Average318.80

Result5 375.06 374.5

RSD (%)0.54 73.66%Average 376.0

Tablet 10Set 4

Potency of standard 99.10% % Weight of standard taken 50.1 mg

Weight of sample taken 210.18 mg Proposed weight / Tablet 210 mg

Factor 1.116

Standard Peak Areas Sample Peak Areas

Sr # Area

RSD(%)

Sr # Area

1 360.5 1 299.1

2 360.5 2 302.2

3 361.1 3 297.7

4 361.6

0.26

Average299.67

Result5 361.1

6 363.0RSD (%)

0.77 73.74%Average 361.3

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Tablet 11Set 5



Factor 1.116


Sr # Area

RSD(%)

Sr # Area

1 363.3 1 302.5

2 365.1 2 302.6

3 365.4 3 302.5

4 366.3

0.30

Average302.53

Result5 363.8

6 364.7RSD (%)

0.02 73.91%Average 364.8

Tablet 12Set 6



Factor 1.116


Sr # Area

RSD(%)

Sr # Area

1 361.6 1 301.5

2 364.4 2 296.9

3 363.7 3 297.0

4 360.4

0.42

Average298.47

Result5 362.0

6 361.2RSD (%)

0.88 72.92%Average 362.2

Average 73.19%RSD 1.40%

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Acceptance Criteria:The % RSD of the assay values should not be greater than 2.0%.

Conclusion:% RSD for assay repeatability is 1.40% (i.e. less than 2.0%) hence repeatability is accepted

3.5.2 Intermediate precisionDIFFERENT DAYS

Tablet 13Day 1



RSD(%)

Sr # Area1 360.5 1 299.12 360.5 2 302.23 361.1 3 297.74 361.6

0.26

Average299.67

Result5 361.16 363.0

RSD (%)0.77 73.74%Average 361.3

Tablet 14DAY 2



RSD(%)

Sr # Area1 365.6 1 304.52 365.7 2 304.33 367.6 3 306.24 368.1

0.28

Average305.00

Result5 366.46 366.3

RSD (%)0.34 74.16%Average 366.6

Absolute difference 0.58 Acceptance Value NMT 4.0

DIFFERENT EQUIPMENTS

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Average 73.95% RSD 0.41%


Tablet 15Agilent (QA-0129)

Potency of standard 99.10% % Weight of standard taken 25.12 mgWeight of sample taken 231.01 mg Proposed weight / Tablet 230 mg

Factor 1.116 Standard Peak Areas Sample Peak Areas

Sr # AreaRSD(%)

Sr # Area1 555.4 1 606.72 551.0 2 608.53 550.2 3 606.54 553.0

0.35

Average607.23

Result5 550.76 551.2

RSD (%)0.18 97.74%Average 551.9

Tablet 16

AGILENT(QA-095)Potency of standard 99.10% % Weight of standard taken 57.94 mgWeight of sample taken 478.07 mg Proposed weight / Tablet 230 mgFactor 1.116


RSD(%)

Sr # Area1 621.1 1 640.52 623.8 2 639.63 626.3 3 642.34 631.3

0.69

Average640.80

Result5 630.3

Average 626.6RSD (%)

0.21 101.26%

Absolute difference 3.54 Average 99.50%

Acceptance Value NMT 4.0 RSD 2.50%

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DIFFERENT ANALYSTTablet 17

Kashif MumtazPotency of standard 99.10% % Weight of standard taken 49.84 mgWeight of sample taken 210.06 mg Proposed weight / Tablet 210 mgFactor 1.116


RSD(%)

Sr # Area1 361.6 1 301.52 364.4 2 296.93 363.7 3 297.04 360.4

0.42

Average298.47

Result5 362.06 361.2

RSD (%)0.88 72.92%Average 362.2

Tablet 18Arslan Khalid



RSD(%)

Sr # Area1 365.6 1 274.12 361.6 2 274.43 362.6 3 274.44 363.3

0.39

Average274.30

Result5 362.66 362.1

RSD (%)0.06 70.51%Average 363.0

Absolute difference -3.35 Average 71.71%

Acceptance Value NMT 4.0 RSD 2.37%

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Acceptance Criteria: The % RSD of the assay/recovery values generated by a single analyst should not be greater than 2.0%. The % RSD of the combined assay/recovery values generated by both analysts, over both days should

not be greater than 3.0%.Absolute difference should not be more than 4.0.

Conclusion:Repeatability has been performed as per protocol and values for % RSDs and Absolute difference is with acceptance limit hence repeatability is accepted.3.6 Limit of Detection (LOD) and Limit of Quantitation (LOQ)LOD was determined by the residual standard deviation and slop of the line in range study.The relationship for Detection Limit is given belowDetection Limit= 3.3* RSD/ mWhere m is the slop of line in linear relation RSD is the residual standard deviation of the y intercept in the linear regression.LOQ was determined by the residual standard deviation and slop of the line in range study.The relationship for Quantitation Limit is given belowQuantitation Limit= 10* RSD/ mWhere m is the slop of line in linear relation RSD is the residual standard deviation of the y intercept in the linear regression

Regression StatisticsR Square 0.999871

Standard Error 2.20688Observations 24

Sum of residual squares 2.20687992

Slop 7.444561404Limit of detection (ppm) 1.0Limit of quantitation (ppm) 3.0

3.7 RobustnessThe robustness of the method was determined to assess the effect of small but deliberate variation of the chromatographic conditions on the determination of Saxagliptin. Robustness was determined by changing the mobile phase pH + 0.2, flow rate to 0.85and 1.15 mL min−1 and the concentration of acetonitrile in the mobile phase to 57 and 63%

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Results from testing of the robustness of the method by changing the pH of the mobile phaseTable 20

Mobile phase pH



Factor 1.116

pH 4.4


Sr # Area

RSD(%)

Sr # Area

1 367.9 1 302.9

2 369.2 2 302.3

3 370.1 3 302.8

4 371.2

0.59

Average302.67

Result5 366.6

6 365.5RSD (%)

0.11 73.24%Average 368.4

pH 4.6


Sr # Area

RSD(%)

Sr # Area

1 365.6 1 304.5

2 365.7 2 304.3

3 367.6 3 306.2

4 368.1

0.28

Average305.00

Result5 366.4

6 366.3RSD (%)

0.34 74.17%Average 366.6

pH 4.8


Sr # Area

RSD(%)

Sr # Area

1 367.6 1 307.1

2 366.8 2 299.7

3 366.6 3 305.1

4 366.5

0.27

Average303.97

Result5 365.9

6 368.7RSD (%)

1.26 73.84%Average 367.0

Average 73.75%

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RSD 0.47%

Acceptable NMT 3%

Results from testing of the robustness of the method by changing the composition of the mobile phase

Table 21Mobile phase Composition

Potency of standard 99.10% % Weight of standard taken 50.23 mgWeight of sample taken 210.13 mg Proposed weight / Tablet 210 mg

Factor 1.116 Acetonitrile 17 %


RSD(%)

Sr # Area1 373.5 1 296.12 373.5 2 296.73 374.5 3 292.84 373.7

0.23Average

295.20Result

5 375.76 374.1

RSD (%)0.71 70.39%Average 373.9

Acetonitrile 20 %Standard Peak Areas Sample Peak Areas

Sr # AreaRSD(%)

Sr # Area1 372.9 1 294.52 372.9 2 293.93 375.8 3 293.04 372.1

0.35Average

293.80Result

5 372.76 373.9

RSD (%)0.26 70.15%Average 373.4

Acetonitrile 23 %Standard Peak Areas Sample Peak Areas

Sr # AreaRSD(%)

Sr # Area1 375.2 2 288.72 377.0 3 289.23 375.7

0.32

Average 288.60 Result4 378.2

RSD (%)

0.23 66.28%

5 376.96 378.8

Average 376.8

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Average 69.61%

RSD 1.16%

Acceptable NMT 3%

Results from testing of the robustness of the method by changing flow rate of the mobile phase

Table 22Mobile phase Flow rate


1.35 mlStandard Peak Areas Sample Peak Areas

Sr # AreaRSD(%)

Sr # Area1 415.8 1 325.02 415.0 2 325.03 415.9 3 324.54 415.7

0.09Average

324.83Result

5 415.56 416.0

RSD (%)0.09 69.67%Average 415.7


Sr # AreaRSD(%)

Sr # Area1 372.9 1 294.52 372.9 2 293.93 375.8 3 293.04 372.1

0.35Average

293.80Result

5 372.76 373.9

RSD (%)0.26 70.15%Average 373.4


Sr # AreaRSD(%)

Sr # Area1 338.9 1 265.82 339.0 2 265.43 339.1 3 265.04 339.3

0.04Average

265.40Result

5 338.96 339.0

RSD (%)0.15 69.79%Average 339.0

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Average 69.87%RSD 0.25%Acceptable NMT 3%

Results from testing of the robustness of the method by changing temperature

Table 23Temperature Change



Factor 1.116

20 ° C


RSD(%)

Sr # Area1 565.5 1 543.12 562.0 2 540.03 563.2 3 543.04 566.9

0.33

Average542.03

Result5 566.36 564.8

RSD (%)0.33 85.56%Average 564.8

25 ° CStandard Peak Areas Sample Peak Areas

Sr # Area

RSD(%)

Sr # Area1 558.9 1 540.72 559.8 2 538.7

3 559.3 3 537.4

4 561.4

1.27

Average538.93

Result5 563.3

6 543.6RSD (%)

0.31 86.15%Average 557.730 ° C


RSD(%)

Sr # Area1 565.0 1 538.52 563.2 2 538.13 563.8 3 539.94 564.6

0.24

Average538.83

Result5 565.66 561.8

RSD (%)0.18 85.17%Average 564.0

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Table 24

Solution stabilityPotency of standard 99.10% % Weight of standard taken 50.33 mg


Factor 1.116 0 Hour


RSD(%)

Sr # Area1 363.3 1 302.52 365.1 2 302.63 365.4 3 302.54 366.3

0.30

Average302.53

Result5 363.86 364.7

RSD (%)0.02 73.94%Average 364.8

24 HourStandard Peak Areas Sample Peak Areas

Sr # AreaRSD(%)

Sr # Area1 365.6 1 304.52 365.7 2 304.33 367.6 3 306.2

4 368.1

0.28

Average305.00

Result5 366.4

6 366.3RSD (%)

0.34 74.46%Average 366.6


3.8 System Suitability

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SYSTEM SUITABILITY

Table 25

Sr. # Peak Area Theoretical plates Symmetry ResolutionCapacity Factor k S/N

1 361.6 5518.0 0.5900 ---- 17.584 126.52 364.4 5480.0 0.5900 ---- 17.397 97.03 363.7 5535.0 0.5900 ---- 17.777 95.34 360.4 5250.0 0.5900 ---- 18.918 89.45 362 5390.0 0.5900 ---- 18.055 89.4

6 361.2 5404.0 0.5900 ---- 18.626 87.1

Results

Peak Area T. P T. F Resolution k S/N

n=6 6 6 6 6 6Mean=362 5430 0.59 ----- 18 97

SD=1.532 105.777 0.000 ----- 0.600 14.717

RSD %=0.4228% 1.9482% 0.0000% ----- 3.3249% 15.1027%

Acceptance CriteriaRelative Standard deviation < 1.0

Theoretical plates > 2000Tailing Factor 0.8 > 2.2

3.9 Conclusion

It is obvious from all the above mentioned results that this method is valid for the analysis of Saglip tablet for intermediate, finished and stability stages. It complies with all the required parameters and hence declared as validated method for Saglip tablet.

3.10 Deviation

The Analytical Method mentioned in protocol was found fit for Saglip Tablet. Diluent was changed (0.1N HCl)and method was VALIDATED thereof (see section 2.1).

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Author, Quality Assurance (Validation), CCL Pharmaceuticals (Pvt.) Ltd Date

Reviewed by, Deputy Manager Quality Assurance (Validation), CCL Pharmaceuticals Pvt. Ltd Date

Reviewed by, QCM, CCL Pharmaceuticals Pvt. Ltd Date

Approved by, Head of QA&R, CCL Pharmaceuticals Pvt. Ltd Date

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analytical method validation report of saxagliptin tablet

Documents

specificity of method

standard solution mgmlcu

sample solution rs

analytical balance

peak response

standard solutioncs

principle peak

validation characteristics