Comparative Dissolution Studies “of Cefxime 400mg capsules. with Protocol No ee | Facility Quality Contro! Department, 1 Cefixime Cefixime (Trihydrate) | 033723 400mg capsules 2 Cefiget 400mg | _ Cefixime (Trihydrate) 441 Getz Pharma (Pvt) Ltd. Capsules.ABI TENT Section ‘Description 1 introduction 2 Objective 3 ‘Scope 4 ‘Analytical procedure and parameter 5 Conclusion 4. INTRODUCTION: Solid, oral-dosage Capsules and Capsules are the most effective and efficient means of treatment available in the pharmaceutical industry. The drug, taken orally, dissolves in the gastrointestinal fluids and becomes available as it is absorbed into systemic circulation, Routine in vivo measurement of the active pharmaceutical ingredient (API) in blood and urine is Not possible in practice. ‘Therefore, in vitro methods to measure the dissolution rate of the API from the solid oral form are officially recognized by regulatory agencies as an important consideration when formulating solid-oral-dosage forms. The biopharmaceutical classification system (BCS) is an important classification used for waiver of in-vivo bioavailability and bioequivalence decisions by regulatory agencies. This classification system is based on the aqueous solubility and intestinal permeability of the API. The system is classified into the following four classes: Class 1: High Solubil ~ High Permeability Class 2: Low Solubility — High Permeability Class 3: High Solubility — Low Permeability Class 4: Low Solubilty ~ Low Permeability Based on low and low permeability, Cefixime is classified as a BCS Class IV drug.The objective of this protocol is to describe the methodology for establishing the similarity of Cefixime 400mg Capsules (Cefixime (Trihydrate) developed in the New Product Development laboratory at Crystolite Pharmaceuticals by comparing it with the dissolution profile of Brand leader of same strength. 3. SCOP! The scope of this protocol is comparison and assessment of dissolution profiles of Cefixime 400mg Capsules with the brand leader. 4, ANALYTICAL PROCEDURE AND PARAMETER: Preparation of buffer for Dissolution: pH4.2 HCI Buffer Dissolve 7.1 ml of Hydrochloric acid (Conc. 12M) and 3.725 g potassium chloride in 1000 mi Purified water and adjust to pH 1.2. Acetate Buffer pH 4.5 Dissolve 4.96 g of sodium acetate anhydrous and 1.6 ml (about) of glacial acetic acid in 1000 ml of purified water and adjust to pH 4.5. Phosphate Buffer pH 6.8 Dissolve 6.8 g of potassium dihydrogen phosphate and 0.9 g of sodium hydroxide in 1000 ml purified water and adjust to pH 6.8. Test Conditions: Apparatus USP type I (Basket) Dissolution Medium 900 mi ‘Sampling Points* 10, 15, 20, 30, 45 & 60 minutes “| Stirring Rate 100 RPM ‘Temperature 37.0% £0.5°Cic System: UV 254nm, Column: 4.6 mm x 12.5 om, 4 um packing L1 Column temp: 40°C Flow rate: Adjusted so that the retention time of Cefixime is 10 minute. Injection volume: 10 pL Mobile Phase: Acetonitrile and solution A (1:3). Solution A: 25m! of 0.4 M tetrabutylammonium hydroxide solution diluted with water to 1000mI, and adjusted with 1.5 M phosphoric acid to a PH of 6.5. Solution B: 13.6g/L of monobasic potassium phosphate in water. Solution C: 14.2g/L of anhydrous dibasic sodium phosphate in water. Diluent: Dissolution medium. Prepare 0.05 Molar Phosphate Buffer having PH 7.2, by taking dibasic sodium phosphate. (DRAP) Procedure for Dissolution: Introduce one Capsule into each dissolution vessel containing 900m! dissolution medium (previously equilibrated at 37°C40.5°C). Run the dissolution apparatus for 60 minutes at 100 RPM. Draw 10 ml of the sample solution at the interval of 10, 15, 20, 30 & 45 ,60 minutes from each bowls separately from the middle between the surface of the test liquid and the upper edge of basket stirrer and at least 10mm away from the vessel wall. Filter @ portion of this solution through 0.22 micron membrane filter and use the filtrate as sample solution. Standard preparation: ‘Transfer an accurately weight quantity of 28mg of Cefixime as working standard into 100ml of volumetric flask & make volume up to the mark with dissolution medium. Sonicate for 15 minutes to Dissolve . Take SmI of the filtrate into 25ml volumetric flask and make volume up to the mark with Dissolution medium. Filter the solution through 0.45p. ‘Sample preparation: Insert 1 Capsule into each of six bowls. Start the procedure. Take 10m! sample from each bow! after 60 minutes. Filter the solution through 0.45.Procedure for Analysis Flush the column with Acetonitrile and water 50:50 ratio at a flow rate of 1.0 ml for 15 minutes and then with water for 30 minutes. Set the conditions outlined under chromatographic system ‘and equilibrate the column with mobile phase until a steady baseline is obtained. After the column is equilibrated, separately inject equal volume of about 10p! of the standard preparation and the sample preparation & record the chromatogram. At the end of the analysis, flush the column with mobile phase for 15 minutes followed by water for 30 minutes and then with Acetonitrile and water 50:50 ratio for 15 minutes. Calculate the quantity of Cefixime by measuring the response for major peak in the sample in percentage by the following formula; Calculatior Result(%) = (ful ta) x (Ce / Cu) x 900 /Cap weight x Average Cap wt / Label Claim x potency Ty = _ peak response of Cefixime from the Sample solution Te = peak response of Cefixime from the Standard solution C. = concentration of Cefixime in the Standard solution (mg/mL) Cy = concentration of Cefixime in the Sample solution (mg/mL) Calculation of Dissimilarity factor f, Use following formula to calculate the value of f, Fh {15 et" | Re Te [ [5ee1” Rd} x 100 Where, n = number of time points, R = Dissolution value of the reference batch at time t, qh = Dissolution value of the test batch at time t. Acceptance Criteria ‘The value of f, should be < 15. Calculation of similarity factor fy Use following formula to calculate the value of fz100 Slewm—-reor = 50-log| I+ n f2 = Similarity Factor N = number of time points Rw = mean % API dissolved of reference product at time point x Tt = mean % AP! dissolved of test product at time point x Acceptance Criteria ‘The value of f, between 50 and 100 suggests that the two dissolution profiles are similar, . Conclusion Conclusion will made after reviewing of results.Benen Comparative Dissolution Studies of Cefixime 400mg capsules with Ceficet | | 400mg Capsules. Report No.: CDP/003/23 Facility: Quality Control Department The comparative dissolution study was performed to establish the comparative dissolution profile of Cefixime 400mg capsules against the Brand leacer product Cefiget 400mg Capsules. Manufactured by Getz Parma. Tre Dissolution Profiling was performed in Test medium 1 (Hydrochloric Acid pH 1.2), Test medium 2 (Acetate buffer pH 4.5) and Test medium 3 (Phosphiite buffer pH 6.8). Dissimilarity factor f1 and similarity factor {2 were calculated ‘or LExécutive Summary | each test medium. In case of Test Medium 1 Cefixime f1= 8.927, 12= 81 105 & V for test medium 2 Cefixime f1= 1.222, f2= 90,649 & for test mediun 3 Cefixime f1= 1.708, f2= 94.628 & have been obtained respectively. Dissimilarity factor f1 and similarity factor £2 have been obtained well within the acceptance criteria for all the 3 test mediums, therefore it is concluded that release profile of test product Cefixime 400mg capsules is in Comparison and Similar with the Reference product of Cefiget 400mg Capsules. TABLE OF CONTENT Section Description Page 7 Objective = 2 Material used and traceability 2 Ail 3 Statistical results of Cefixime 400mg Capsules Vs. Cefiget 400mg Caps 3 Graphical Representation of Cefixime 400mg capsules with Cefiget 4 21 pn 400mg Capsules results. Conclusion 241. OBJECTIVE: To compare the dissolution a) pH 1.2 Hydrochloric Acid Buffer b) pH4.5 Acetate Buffer ©) pH 6.8 Phosphate Buffer 2. MATERIAL USED AND TRACEABILITY: Name Cefixime Potency of Working Standard Cefixime (Trihydrate) 88.6% (ODB) and (99.1% on anhydrous basis Lac No. 320/RMI23 aed Batch Name Number Date Manufactured By Cefixime 400mg capsules 033723 09-2025 Cefiget 400mg Capsules 441 3.2025 | Getz Pharmaceuticals Name Lot NumberiB# Manufacturer Hydrochlorie Acid 52385917 Sigma Aldrich ieee aihydrogen | 90,4146, 1000 Chem-lab Phosphate Sodium hydroxide $0181UF2 DAEJUNG ‘Sodium Acetate Anhydrous ‘S0466TK1 DAEJUNG [Phosphoric Acid POOOSEQNI DAEJUNG ethanol (Analytical Grade). | CLO0.1363.2500 Chem-lab /Name_ Hea tr HPLC ‘Shimadzu SPD-20A CTIEQIQC/007 Dissolution Apparatus QUARTZ -74TS CTIEQP/QCIO29 pH Meter INOLAB-7110 CTIEQPIQCING Analytical Balance ATX 224 CTIEQP/QCIO10 Profile of Cefixime 400mg capsules with Cefiget 400mg Capsules for - API_Cefixime (Trihydrate) as per protocol CDP/003/2023 in the following dissolution media:STATISTICAL RESULTS: 3.4 pH 41.2 HCI Buffer Time Duration: 10 minutes Time Duration: 15 minutes No. of : Cefiget No. of samples | Cefixime 400mg samples | Cefixime Cefiget 400mg capsules 400mg 400mg Capsule Capsule aa Cofixime 1° Cofixime Cefixime 1 13.90% 11.30% 1 15.90% 7.40% 2 12.92% 12.43% 2 20.95% 18.23% 3 12.40% 11.90% 3 18.40% 18% 4 12.33% 11.73% 4 18.35% 16.62% 5 12.50% 11.60% 5 20.40% 7.10% 6 12.34% 12.79% 6 18.66% | 17% 7 13.30% 12.70% 7 18.20% 16.90% 8 12.88% 12.60% 8 18.05% 26.90% 9 13.30% 12.30% 9 15.00% 17.40% 10 13.10% 12.87% 40 15.57% 18.14% 44 13.10% 11.10% 41 21.30% 118% 12 12.54% 12.09% 42 15.37% 8.25% Average 12.88% 12.12% ‘Average 18.01% 37.50% ‘STDEV 0.004839132 | _0.00594767 ‘STDEV 0.021813887 | 0.005¢76545 % RSD 3.755874917 | 4.908330531 % RSD 12.1104163 |_3.41614465 Min 12.33 11.10% ‘Min 15.00% 46.62% Max. 13.9 12.87% Max 21.30% 13.25%Time Duration: 20 minutes Time Duration: 30 minutes Cefixime Cefixime ; No. of | Capeuie | Safget No. of Capeute | Setaet samples Capsule ‘samples. Capsule Cefixime Cefixime Cefixime Cefixime - 25.70% 15.90% 7 18.30% 48.00% 2 22.61% 14.68% 2 18.47% 78.69% 3 21.60% 15.70% 3 18.00% 21.30% 4 21.74% 23.02% 4 18.00% 49.23% 5 21.20% 16.40% 5 17.90% 48.90% 6 19.07% 14.21% 6 18.41% 48.50% fe 17.50% 13.70% 7 16.80% 18.00% 8 17.97% 13.78% 8 16.85% 47.76% 9 18.40% 13.70% 9 17.70% 48.60% 10 18.12% 13.57% 10 16.66% 20.55% Lu 18.30% 13.40% 44 17.80% 8.30% 2 1e.10%| 14.60% 12 19.89% 18.46% Average 20.03% 15.22% Average 17.90% £8.86% STDEV 0025253603 | 0,026517673 STDEV 0,008854976 | 0.010590315 %RSD 12.61051297 | 17.42100478 %RSD__ | 4.947374808 | 5.615°69797 Min 17.50% 13.40% Min 16.66% 37.76%. Max 25.70% 23.02% Max’ 19.89% 2.30%Time Duration: 45 minutes Time Duration: 60 minutes Cefixime Cefixime No.of Gapoute | Saree No. of| Gapoure | Saget samples Capsule samples Capsitle Cefixime | Cefixime Cefixime | Cefixime = 23.40% 21.20% 1 22.70% 41.60% 2 23.88% 21.63% 2 22.93% 45.57% 3 23.30% 20.90% 3 22.20% 4 23.26% 19.96% 4 22.26% 5 20.80% 20.80% 5 22.50% 6 21.34% 20.31% 6 22.49% 7 20.60% 19.50% Li 21.90% 8 23.01% 20.04% 8 22.06% 9 20.50% 21.00% 9 22.00% 10 22.71% 23.05% 10 20.27% 21.00% a1 22.80% 19.80% 1 21.50% 49.40% 12 21.01% 20.88% 12 18.48% 22.28% Average 22.22% 20.76% Average 21.77% 12.85% STDEV 0.012589398 | 0.009609508 STDEV 0.012427057 | 0.026#10921 % RSD 5.666433441 | 4.629786835 %RSD__ | 5.707247978 Min 20.50% 19.50% Min 18.48% Max 23.88% 23.05% Max 22.93% }Time “aoomg. | Saget n0mg SNo eee (Cettxime) | (Cefxime) Ty | cRw-rO? (minutes) Be After 10 12.880 12.120 0.760 0.578 2 After 15. 18.010 17.500 0.510 0.260 3 | After 20 20.030. 15.220 4.810. 23.136. = After 30 17.900 18.860 0.960 0.922 5 After 45, 22.220 20.760. 1.460 2.132 6 After 60 21.770 22.850. 1,080 1,166 SUM 107.310 9.580. 28.193 14+9.121/5 5.699 SORT 2.520 2.387 S0ieg 097587 sits 8.927 81.1053.2 pH 4. Acetate Buffer Time Duration:10minutes Time Duration:15minutes No. of | Cefixime Cefiget Cefixime | Cefiget, samples | 2209 400mg No. of samples 400mg 400mg , ES Capsule Capsule Capsule Capsul> Cefixime Cefixime Cefixime —_| Cefixine 1 38.00% 36.80% 1 62.30% 66.70% 2 36.84% 36.65% 2 66.62% 61.98% 3 36.70% 37.30% 3 66.50% 61.40% 4 36.40% 36.29% 7 65.94% 66.11% 5 36.70% 37.30% 5 66.40% 66.20% 6 36.66% 37.35% 6 66.57% 63.52% 7 36.40% 37.20% 7 66.20% 61.00% 8 36.59% 37.34% 8 66.40% 6.42% 9 37.30% 37.30% 9 66.20% 66.20% 10 36.28% 38.58% 10 65.75% 69.71% 44 37.30% 37.60% 4 66.20% 61.20% 12 36.34% 37.57% 12 65.90% 6!.86% Average 36.79% 37.27% ‘Average 65.92% 61.69% STDEV 10.00506928 | 0.005640331 STDEV 0.011708078 | 0.0104 8649 % RSD 1377775497 | _1.51323492 % RSD 1.776238795 |_1.5682(9232 Min 36.28% 36.29% Min 62.30% 61.86% Max 38.00% 38.58% Max. 66.62% 62.71%Time Duration: minutes 30 No. of | Cefixime Cefiget Cefixime Cefiget, samples | 420mg 400mg No. of samples 400mg 400mg® Capsule Capsule Capsule Capsul> Cofixime Cofixime Cefixime | Cefi 1 92.80% 80.10% 1 97.10% 97.10% 2 91.71% 91.06% 2 96.30% 93.75% 3 91.70% 90.50% 3 95.80% 98.80% 4 91.39% 89.87% 4 92.50% 5 90.30% 90.40% 5 96.30% 93.10% 6 90.65% 90.73%, 6 95.90% 91.15% 7 90.20% 85.40% 7 95.80% 95.40% 8 90.68% 87.84% 8 96.10% 9.01% 9 90.30% 89.90% 9 95.90% 9.70% 10 89.79% 89.39% 10 95.40% 6% 4 90.40% 89.90% 4 96.10% 91..30% 12 91.26% 89.40% 12 95.50% 98.38% Average 90.93% 88.71% Average 95.73% 98.48% STDEV| _0.008563541 | 0.031133791 STDEV 0.011062837 | 0.0101¢4987 % RSD 0.941755692 | 3.509713519 % RSD 1.155689428 | 1.064660964 Min 89.79% 80.10% Min 92.50% 93.75% Max 92.80% 91.06% Max 97.10% 10% i 3 5Time Duration: 45 Time Duration: 60 ninutes minutes No. of | Cefixime Cefiget Cefixime samples | 420mg 400mg No. of samples 400mg Capsule Capsule Capsule Cefixime Cefixime Cefixime Cefixinie co 1 90.90% 94.50% 4 101.90% 93.10% 2 94.50% 92.68% 2 101,30% 92.27% 3 94.20% 92.10% 3 93.30% 98.60% 4 93.80% 95.23% 4 98.00% 95.48% 5 94.30% 94.10% 5 92.10% 98.20% 6 94.60% 95.83% 6 93.90% 99.33% a 94.20% 95.60% 7 97.50% 95:90% 8 94.70% 96.04% 8 98.00% 97.88% 9 94.50% 93.90% 9 94.30% 94.00% 10 94.00% 95.29% 10 93.80% 97.17% 14 92.90% 95.70% 14 94.20% 98.80% 12 93.90% 95.15% 12 97.30% 97.99% ‘Average 93.88% 94.68% Average 96.30% 9.98% STDEV| _0.010532417 | 0.012106426 STDEV 0.031857638 | _0.022%8093 ie % RSD 1,121961916 | 1.278712764 % RSD 3,308165927 | 2.409170686. Min 90.90% 92.10% Min 92.10% 92.27% Max. 94.70% 96.04% Max. 101.90%ind Dissimilarity Factor for Cefixime al pHa : ‘oa | ValrtS0mg | Savesto. 50mg e - to ere Seu | eSteunnay [8° | emvoren? (minutes) | %Release T(t) | %Release Rit) 1 [After 10 36.790 37.270 0.480 | 0.230 2 | After 15” 65.920 68.690 0.770 0.593 3 | After 20 90.930 88.710 2220 | 4.926 4 | After 30 95.730 96.480 0.250 | 0.063 5 [After 45 93.880 94.680 0.800 | 0.640 8 [After 60 96.300 94.980 1320 | 1.742 ‘suM 477.ei0__[ 5.840 | 2.107 1#230,42018 2.366 SQRT 39.403, 1.538 S0log 10016.277 90.649 | f1 Value 4.222 90.649, L3.3 pH 6.8 Phosphate Buffer Time Duration: 10 minutes Time Duration: 15 minutes Cefixime | Cefiget Cefixime | cefiget No. of | 400mg 400mg No. of | 400mg 400mg samples | Capsule | Capsule samples | Capsule | Capsule Cefixime _ | Cefixime Cefixime | Cefixime = 76.90% 75.40% 2 92.20% 89.80% 2 76.54% 74.93% 2 88.38% 89.36% 3 76.80% 79.20% | [3 88.80% 87.10% 4 72.82% 74.84% | [4 88.09% 36.44% 5 72.70% 7a40%| [5 88.30% 86.30% 6 73.32% 74.90% | [6 88.7% 86.78% ee 71.60% 74.90% - 88.40% 85.00% 8 72.63% 74.68% 8 86.56% 87.22% 9 73.40% 74.10% 9 87.60% 87.00% 10 2.72% 7asi%| [10 86.88% 87.65% 1 73.30% 74.80% | [44 91.20% 86.80% 12 72.56% 7ao1%| [42 87.16% 85.80% Average 3.77% 75.11%| | Average 88.53% 87.10% STDEV | 0.018545593 | 0.015600857| | STDEV 0.016613074 | 0.013515073 %RSD_| 2513832919 | 207718308] [% RSD 1.876582764 | 1551598904 Min 71.60% 74.01% Min 86.56% 85.00% Max 76.90% 79.80% Max 92.20% 89.80%‘Caleulation|of Simi arity and Bissim Cef = {time “me Ceti 00mg ; | Pero Ganstiee | Geeaues Tawa | cxo-n0) (minutes) | %Release T(t) %Release Rit) 4 After 10 73.770. 75.110 1.340 1,796 2 After 15. 88.530 87.100. 1.430 2.045 SUM. 162.210 2.770 3.841 1+68.330/5, 1.640 SQRT 12.388 4.281 50log 100/3.520 94.628 1.708 94.628 4. Graphical Representation 4.1 pH 1.2 HCI Buffer Acceptance criteria: Calculate f1 value and 2 value. If f1 value s 15 and {2 > 50, the profiles are regarded as. similar and no further in vivo studies are necessary, Result Cefixime: (F1 = 8.927, f2 = 81.105%) Remarks: Cefixime 400mg Capsule complies with respect to the acceptance criteria of f1 and f2, when compared against the brand leader product using dissolution medium pH 1.2.28.000 7 20.000 15.000 4 —eCefixime ‘ 90.000, J} $$ ae cofiget 500 eee 0.000 $$$, 0 1 2 30 4 so 6 7 ee aes 4.2pH4.5 Acetate Buffer ——— 820, a Cp ee ——————— 60 + o—Cefixime : Be cefiget 40 + —E 20 pe 0} ° 20 40 60 80 Acceptance criteria: Calculate f1 value and f2 value. If f1 value s 15 and {2 > 50, the profiles are regarded as similar and no further in vivo studies are necessary. Result Cefixime: (F1 = 1.222, f2 = 9.649%) Remarks: A Cefixime 400mg capsule complies with respect to the acceptance criteria of f1 and £2, when compared against the brand leader product using dissolution medium pH 4.5.Result Cefixime: (f1 = 1.222, f2 = 90.649%) Remarks: A Cefixime 400mg capsule complies with respect to the acceptance criteria of f1 and £2, when compared against the brand leader product using dissolution medium pH 4.5. 4,3 pH 6.8 Phosphate Buffer —eCefixime ~acefiget Acceptance criter Calculate f1 value and 12 value. If f1 value < 15 and f2 > 60, the profiles are regarded as similar and no further in vivo studies are necessary. Result Cefixime: (ft .708, £2 = 94.628%) Remarks:Cefixime 400 Capsules complies with respect to the acceptance criteria of f1 and f2, when compared against the brand leader product using dissolution medium pH 6.8, 5. Conclusion Dissimilarity factor f1 and similarity factor £2 have been obtained well within the acceptance criteria for all the 3 test mediums, therefore it is concluded that release Profile of test product Cefixime 400 Capsules is Comparatively Similar with the reference product Cefiget 400mg Capsules,