Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 3, No. 1, 4-7 2019 DOI: 10.33805/2576.8484.160 © 2019 by the authors © 2019 by the authors History: Received: 15 November 2018; Accepted: 19 December 2018; Published: 1 January 2019 * Correspondence: shivajij@indoco.com Development and Validation of New Analytical Method for the Determination of Particle Size Distribution in Glimepiride Using Laser Based Particle Size Analyzer Gosar A1, Jadhav S1*, Patil V1, Folane S1, Jadkar A1, Vispute T1 1Indoco Remedies LTD R&D Centre, Navi Mumbai, India; shivajij@indoco.com (J.S.). Abstract: Particle size is a key factor for modern drug quality since it affects the bioavailability and dissolution profile of the drug product. Study for Particle size is helpful to optimize drug product development process and improve the quality of drugs. In Order to determine the particle size of Glimepiride a novel and accurate Particle size determination method has been developed for the determination of particle size distribution of Glimepiride was described in this paper. This method has shown good reproducible results. By using water as dispersant wet method is developed and validated as per International conference on Harmonization guidelines (Q2 (R1)) and found out robust and reproducible with % RSD of d (10), d (50) and d (90) values found within accepta nce limit ranges from 6.05% to 21.84% for d(0.1), 3.04% to 9.87% for d(0.5) and 4.69% to 14.32% for d(0.9) in validation. The described method is accurate and validated and successfully applied for the determination of particle size distribution of Glimepiride. Particle size method is discussed in detail to ensure in-depth understanding of particle size distribution and particle size method performance during lifetime of the product. Keywords: Glimepiride, Method development, Method validation, Particle size analyzer. 1. Introduction Glimepiride is a white to yellowish-white, odorless powder and is practically insoluble in water. Glimepiride acts as an insulin secretagogue and it lowers blood sugar by stimulating the release of insulin by pancreatic beta cells and by inducing increased activity of intracellular insulin receptors [1]. Glimepiride chemically, is 3-ethyl-4-methyl-N-[2-[4-[(4- methylcyclohexyl)carbamoyllsulfamoyl] phenyl] ethyl]-2-oxo-5H-pyrrole-1-carboxamide is a third generation sulfonylurea derivative which is commonly used in the treatment of non-insulin dependent Type 2 diabetes mellitus [2-5]. Glimepiride, marked under the trade name Amaryl, is the first line medication for the treatment of type-2 diabetes mellitus [6-8]. Relative bioavailability, extended-released along with bioequivalence and immediate released is related to particle size of Glimepiride [9-12]. Literature survey done for Particle size distribution method for Glimepiride and found that particle size determination method is not available. Therefore, study was carried out to develop a method to determine particle size distribution of Glimepiride by Particle size analyzer and further validation of the method, was carried out. 2. Materials and Reagent 2.1. Reagents Triton x-100, water and Glimepiride was obtained from Indoco Remedies Ltd, Navi Mumbai, India. 2.2. Instrumentation Particle size analyzer system of make Malvern and model is 2000 consists of Dry Scirocco 2000 and wet Hydro2000S accessories. 3. Results and Discussion 3.1. Analytical Method Development The main goal was to develop method to obtain the most stable, reproducible, reliable method. Various dispersant were tried for the Glimepiride on the basis of solubility. Water is found well suitable dispersant to develop method for determination of particle size distribution of Glimepiride. Various trial were taken during method development as mention below, Glimepiride solubility within a suitable solvent system a critical role in particle size wet method development since sample preparation is play vital role. Make Malvern Mastersizer, Model Mastersizer 2000, Dispersion Unit Scirocco 2000, Method parameters are kept as Particle Refractive index is 1.520, Absorption is 0.1, Analysis Model General purpose, Obscuration range is 1% to 6%, Dispersive Air pressure is 2.0 bar, Vibration feed rate is 40%, Sensitivity is Normal, Sample measuring time 5 seconds, Background measuring time 5 seconds, sample preparation is Transferred about 1 to 2 g of sample into the sample tray with the help of a cleaned spatula and carried out the analysis The obscuration values are well within the limit and also weighted residual is less than 1%. Hence, proceeded for next trial by changing the feed rate 45% to check the effect on particle size. Various trials were taken by changing instrumental parameters. The obtained results of d(10), d(50) and d(90) values are not reproducible and % RSD is also not well within general criteria of USP, General Chapter <429>. Hence next development was carried out by wet analysis. Method parameters for wet analysis is Equipment Malvern Mastersizer, Model Mastersizer 2000, Sample handling unit is Wet Dispersion Unit, Sample model is Hydro 2000S, Dispersant name is Water, Dispersant refractive index is 1.330, Sample refractive index is 1.520, Sample absorption is 0.1, Sample measurement time is 10 second, Measurement Snaps is 10,000. Background Measurement time is 10 second, Background Snaps 10,000, Obscuration range is 10-30%, Stirrer speed is 2000 rpm, No. of measurement cycle 03,sample preparation is Weighed 200.26 mg of sample and transferred in beaker added 1-2 drops of Triton x-100 and few drops of dispersant and make a paste by using glass rod, then added 20 mL of dispersant. Sonicate externally for 40 seconds to form homogeneous solution. Sample solution was added in the dispersion unit when “Add sample under Obscuration” message was shown by the instrument software and performed the analysis. All method parameters were kept as per above only changed in sonication time 50 Sec. and performed the analysis. The obscuration is well within limit it is proved that there is no any effect of sonication time on particle size, Hence, preceded for next trial by changing stirrer speed check the effect on particle size. The obscuration values are well within the limit, from the obtained results there is negligible effect on the particle size by decreased in stirrer speed. Hence, Malvern Mastersizer, Model Mastersizer 2000, Sample handling unit is Wet Dispersion Unit, Sample model is Hydro 2000S, Dispersant name is Water, Dispersant refractive index is 1.330, Sample refractive index is 1.520, Sample absorption is 0.1, Sample measurement time is 10 second, and Measurement Snaps is 10,000. Background Measurement time is 10 second, Background Snaps 10,000, Obscuration range is 10-30%, Stirrer speed is 1800 rpm, No. of measurement cycle 03. Performed analysis repeatability with weight 200.68, 200.96 mg and 200.31 mg to check consistency. The obtained results of d(10), d(50), and d(90) values of analysis in twice are close to each other, weighted residue is less than 1% and obscuration is found satisfactory. To check the reproducibility of method three replicate of sample was analyzed and results were found reproducible & percentage Relative standard deviation of d(10), d(50), and d(90) is respectively 4.27%, 9.83% and 14.45 %. 3.2. Method Validation The validation work was conducted according to the ICH (International Conference on Harmonization) guidelines Q2R1. The method validation parameters include Precision, Intermediate Precision, Robustness and Batch Analysis. mailto:shivajij@indoco.com 5 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 3, No. 1: 4-7, 2019 DOI: 10.33805/2576.8484.160 © 2019 by the authors 4. Method Validation Parameters 4.1. Method Precision Procedure: Determined the particle size of six precision samples as per above method and recorded the particle size for d(0.1), d(0.5) and d(0.9) in Table 1. Table 1. Method precision. Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Precision-1 2.553 15.654 64.835 Precision-2 2.697 17.513 76.598 Precision-3 2.77 18.122 76.865 Precision-4 2.578 16.981 63.762 Precision-5 2.897 17.489 83.82 Precision-6 2.766 17.871 74.388 Average 2.71 17.272 73.378 % RSD 4.78 5.11 10.52 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 4.2. Intermediate Precision Procedure: Determined the particle size of six Intermediate precision samples as per above method and recorded the particle size for d(0.1), d(0.5) & d(0.9) particles in Table 2 and similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of twelve measurements i.e. six of precision and six of Intermediate precision. Table 2. Intermediate precision. Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Intermediate precision-1 2.725 17.744 79.323 Intermediate precision-2 2.624 17.172 75.362 Intermediate precision-3 2.486 17.48 70.604 Intermediate precision-4 2.75 18.339 80.652 Intermediate precision-5 2.784 18.686 82.782 Intermediate precision-6 2.795 18.057 74.775 Average 2.694 17.913 77.25 %RSD 4.41 3.12 5.8 Precision-1 2.553 15.654 64.835 Precision-2 2.697 17.513 76.598 Precision-3 2.77 18.122 76.865 Precision-4 2.578 16.981 63.762 Precision-5 2.897 17.489 83.82 Precision-6 2.766 17.871 74.388 Cumulative average 2.702 17.592 75.314 Cumulative % RSD 4.4 4.43 8.43 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 5. Robustness 5.1. Robustness-1 (Change in Stirrer Speed to 1800 rpm) Procedure: Determined the particle size of three replicates as per method of analysis, only change the stirrer speed to 1800 rpm and recorded particle Size for d (0.1), d(0.5) & d(0.9) in Table 3 and similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of nine measurements i.e. six of precision and three of Robustness-1. Table 3. Robustness-1 (Change in stirrer speed to 1800 rpm). Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Robustness-1(1) 2.783 18.529 80.054 Robustness-1(2) 2.925 19.554 78.597 Robustness-1(3) 2.901 19.423 80.885 Average 2.87 19.169 79.845 % RSD 2.65 2.91 1.45 Precision-1 2.553 15.654 64.835 Precision-2 2.697 17.513 76.598 Precision-3 2.77 18.122 76.865 Precision-4 2.578 16.981 63.762 Precision-5 2.897 17.489 83.82 Precision-6 2.766 17.871 74.388 Cumulative average 2.763 17.904 75.534 Cumulative % RSD 4.9 6.76 9.18 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 5.2. Robustness-2 (Change in Stirrer Speed to 2200 rpm) Procedure: Determined the particle size of three replicates as per method of analysis, only change the stirrer speed to 2200 rpm and recorded the particle Size for d(0.1), d(0.5) & d(0.9) in Table 4 and similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of nine measurements i.e. six of precision and three of Robustness-2. 6 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 3, No. 1: 4-7, 2019 DOI: 10.33805/2576.8484.160 © 2019 by the authors Table 4. Robustness-2 (Change in stirrer speed to 2200 rpm). Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Robustness-2(1) 2.81 18.692 77.187 Robustness-2(2) 3.23 17.95 89.245 Robustness-2(3) 3.122 17.852 89.001 Average 3.054 18.165 85.144 % RSD 7.14 2.53 8.09 Precision-1 2.553 15.654 64.835 Precision-2 2.697 17.513 76.598 Precision -3 2.77 18.122 76.865 Precision -4 2.578 16.981 63.762 Precision -5 2.897 17.489 83.82 Precision -6 2.766 17.871 74.388 Cumulative average 2.825 17.569 77.3 Cumulative % RSD 8.07 4.89 11.84 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 5.3. Robustness-3 (Change in Obscuration Range to 10-20%) Procedure: Determined the particle size of three replicates as per method of analysis, only change the obscuration range to 10-20% and recorded the particle size for d(0.1), d(0.5) & d(0.9) in Table 5 and Similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of nine measurements i.e. six of precision and three of Robustness-3. Table 5. Robustness-3 (Change in Obscuration range to 10-20%). Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Robustness-3(1) 2.838 19.476 85.459 Robustness-3(2) 3.179 18.953 95.103 Robustness-3(3) 2.767 18.475 79.326 Average 2.928 18.968 86.629 % RSD 7.52 2.64 9.18 Precision -1 2.553 15.654 64.835 Precision -2 2.697 17.513 76.598 Precision -3 2.77 18.122 76.865 Precision -4 2.578 16.981 63.762 Precision -5 2.897 17.489 83.82 Precision -6 2.766 17.871 74.388 Cumulative average 2.783 17.837 77.795 Cumulative % RSD 6.67 6.31 12.66 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 5.4. Robustness-4 (Change in Obscuration Range to 20-30%) Procedure: Determined the particle size of three replicates as per method of analysis, only changed the obscuration range to 20 – 30 % and recorded the particle size for d(0.1), d(0.5) & d(0.9) in Table 6 and similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of nine measurements i.e. six of precision and three of Robustness-4. Table 6. Robustness-4 (Change in Obscuration range to 20-30%). Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Robustness-4(1) 2.304 17.958 79.025 Robustness-4(2) 2.778 18.596 82.875 Robustness-4(3) 2.869 19.828 83.88 Average 2.65 18.794 81.927 % RSD 11.45 5.06 3.13 Precision -1 2.553 15.654 64.835 Precision -2 2.697 17.513 76.598 Precision -3 2.77 18.122 76.865 Precision -4 2.578 16.981 63.762 Precision -5 2.897 17.489 83.82 Precision -6 2.766 17.871 74.388 Cumulative average 2.69 17.779 76.228 Cumulative % RSD 6.89 6.39 9.92 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 5.5. Robustness - 5 (Change in Sample Measurement Time to 9 Seconds from 10 Seconds) Procedure: Determined the particle size of three replicates as per method of analysis, only changed the sample measurement time to 3 seconds and recorded the particle size for d(0.1), d(0.5) & d(0.9) in Table 7 and similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of nine measurements i.e. six of precision and three of Robustness-5. 7 Edelweiss Applied Science and Technology ISSN: 2576-8484 Vol. 3, No. 1: 4-7, 2019 DOI: 10.33805/2576.8484.160 © 2019 by the authors Table 7. Robustness - 5 (Change in sample measurement time to 9 seconds from 10 seconds). Sample ID Particle size (µm) d(0.1) d(0.5) d(0.9) Robustness-5(1) 2.801 19.021 78.032 Robustness-5(2) 2.837 18.746 76.702 Robustness-5(3) 2.738 18.462 77.897 Average 2.792 18.743 77.544 % RSD 1.79 1.49 0.94 Precision-1 2.553 15.654 64.835 Precision-2 2.697 17.513 76.598 Precision-3 2.77 18.122 76.865 Precision-4 2.578 16.981 63.762 Precision-5 2.897 17.489 83.82 Precision-6 2.766 17.871 74.388 Cumulative average 2.737 17.762 74.767 Cumulative % RSD 4.13 5.76 8.64 Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 5.6. Robustness - 6 (Change In Sample Measurement Time to 11 Seconds from 10 Seconds) Procedure: Determined the particle size of three replicates as per method of analysis, only changed the sample measurement time to 7 seconds and recorded the particle size for d(0.1), d(0.5) & d(0.9) in Table 8 and similarly, calculated cumulative average and cumulative percent relative standard deviation for particle size at d(10), d(50) & d(90) of nine measurements i.e. six of precision and three of Robustness-6. Acceptance criteria: The % RSD d(10), d(90) particle size values should not be more than 15, d(50) particle size values should not be more than 10. If the particle size is below 10 µm then the % RSD of d(10), d(50), and d(90) will be doubled. 6. Conclusion Method for determination of particle size distribution of Glimepiride was developed and validated using Laser diffraction technique. Dry dispersion technique was assesses and found that dry dispersion technique is not suitable for and wet dispersion was explored during development trials. Wet dispersant method frozen for determination of particle size distribution of Glimepiride. In method validation, method was found précised with % RSD of 10.59% for d(0.1), 8.57% for d(0.5) and 14.32% for d(0.9). In intermediate precision % RSD obtained were 6.05% for d(0.1), 3.04% for d(0.5) and 5.95% for d(0.9). Also, Cumulative % RSD obtained were 8.39% for d (0.1), 6.71% for d (0.5) and 5.12.40% for d (0.9).Hence, the method considered as rugged. certain parameters were modified within the allowed range and the results obtained were within the acceptance criteria and % RSD ranges from 6.22% to 21.84% for d(0.1), 3.47% to 9.87for d(0.5) and 4-.69% to 12.85% for d(0.9). It proved method is rugged. All the analytical data of development and validation has been compiled and found to be satisfactory. Hence, method developed for the particle size method can be suitably used for analysis of Glimepiride active pharmaceutical ingredient. Table 8. Robustness - 6 (Change in sample measurement time to 11 seconds from 10 seconds). Sample no. Particle size (µm) d(0.1) d(0.5) d(0.9) Robustness-6(1) 2.644 16.327 68.769 Robustness-6(2) 2.638 16.396 65.908 Robustness-6(3) 2.591 16.443 73.409 Average 2.624 16.389 69.362 % RSD 1.11 0.36 5.46 Precision-1 2.553 15.654 64.835 Precision-2 2.697 17.513 76.598 Precision-3 2.77 18.122 76.865 Precision-4 2.578 16.981 63.762 Precision-5 2.897 17.489 83.82 Precision-6 2.766 17.871 74.388 Cumulative average 2.682 16.977 72.039 Cumulative % RSD 4.18 4.86 9.3 References [1] S. N. Davis, "The role of glimepiride in the effective management of type 2 diabetes," Journal of Diabetes and its Complications, vol. 18, no. 6, pp. 367-376, 2004. [2] T. 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