A NEW AND RAPID ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF METFORMIN, PIOGLITAZONE AND GLIMEPRIDE IN TABLET DOSAGE FORM BY USING UPLC

Abstract

The present work was undertaken with the aim to develop and validate a rapid and consistent UPLC method in which the peaks will be appear with short period of time as per ICH Guidelines. The proposed method was simple, fast, accurate and precise method for the Quantification of drug in the dosage form, bulk drug as well as for routine analysis in Quality control. UPLC method was developed and validated for simultaneous estimation of Metformin, Pioglitazone and Glimepiride in bulk drug and in combined dosage forms. The UPLC separation was achieved on a Symmetry C18 (2.1 x 100 mm, 1.7 mm, Make: BEH) or equivalent in an Isocratic Mode. The mobile phase was composed of Phosphate Buffer (25 %) whose pH was adjusted to 4.3 by using Ortho Phosphoric Acid and Methanol (75 %) [UPLC Grade] The flow rate was monitored at 0.25 ml per min. The wavelength was selected for the detection was 258 nm. The run time was 5 minutes. The retention time found for the drugs Metformin, Pioglitazone and Glimepiride were 0.002 minutes, 1.773 minutes and

2.409 minutes respectively. The % recovery was found to be 99.7 % - 100.9 % for the drug Metformin. The % recovery was found to be 98.4 % - 100.9 % for the drug Pioglitazone. The % recovery was found to be 99.9 % - 101.2 % for the drug Glimepiride. The linearity was established in the range of 400 to 600 ppm for the drug Metformin and 12 to18 ppm for the drug Pioglitazone and 0.8 to 1.2 ppm for the drug Glimepiride. The LOD for the drugs Metformin, Pioglitazone and Glimepiride were found to be 0.12 µg/ml, 0.002 µg/ml and 0.002 µg/ml respectively. The LOQ for the drugs Metformin, Pioglitazone and Glimepiride were found to be 0.45 µg/ml, 0.15 µg/ml and 0.08 µg/ml respectively. The proposed method was adequate sensitive, reproducible, and specific for the determination of Metformin, Pioglitazone and Glimepiride in bulk as well as in Tablet dosage form. The validation of method was carried out utilizing ICH-guidelines. The described UPLC method was successfully employed for the analysis of pharmaceutical formulations containing combined dosage form. Overall the proposed method was found to be suitable and accurate for the Quantitative determination of the drug in Tablet dosage form. The method was simple, precise, accurate and sensitive and applicable for the simultaneous determination of Metformin, Pioglitazone and Glimepiride in bulk drug and in combined dosage forms.