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ISSN 2063-5346
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DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF EMPAGLIFLOZIN, LINAGLIPTIN AND METFORMIN

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Kalpesh C Raka, Dr. Kavita R Loksh
» doi: 10.31838/ecb/2023.12.4.082

Abstract

Analytical method development and validation are the continuous and inter-dependent task associated with the research and development, quality control and quality assurance departments. The analytical method development is the need of pharmaceutical industries. The development of methods usually requires the collection of method specifications and the decision on the type of instrumentation. Diabetes is a metabolic disorder with major complication associated with hyperglycemia. India is deemed as the world's capital of diabetes. The diabetic population in the country is close to hitting the alarming mark of 69.9 million by 2025 and 80 million by 2030. Combinations of various anti-diabetics are used for better sugar control. Nowadays various new combinations are in evaluated & are approved for better sugar control such as Empagliflozin, Linagliptin & Metformin hydrochloride. This study describes the development of a rapid, precise, selective and sensitive reverse phase high-performance liquid chromatography method (RP–HPLC) for the quantitative simultaneous determination of Empagliflozin, Linagliptin and Metformin and validation as per International Council of Harmonization (ICH) guidelines. Initial identification of API was performed and is obaestic point was determined using UV spectrophotometry. In the present work, good chromatographic separation was achieved by isocratic method using aC18 (COSMOSIL)250 mm length x 4.6 mm internal diameter, 5 m and a mobile phase consisting of Methanol: 0.1 % OPA Water (30:70)at a flow rate of 0.7 ml/min. G-13148 (DAD) detector at 224 nm was used for detection. The calibration curves obtained were linear (r2=0.999) over the concentration range of 2-10 μg/ml, 0.4 – 2 μg/ml and 80-400 μg/ml for Empagliflozin, Linagliptin and Metformin respectively. The retention time of Empagliflozin, Linagliptin and Metformin was found to be about 6.1038min, 9.4754and 3.4846min respectively. The percentage (%) recovery was found within range for Empagliflozin, Linagliptin and Metformin hydrochloride. As per the international Conference on Harmonisation (ICH, Q2) guideline, proposed RP–HPLC method validation has been carried out. The proposed RP–HPLC method was repeatable and selective as per statistical analysis and it can be used for simultaneous estimation of Empagliflozin, Linagliptin and Metformin hydrochloride

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