Comparative Studies on Simultaneous Estimation of Metformin, Empagliflozin, Remogliflozin, Glimepride and Gliclazide by using RP-HPLC and Diol Technique

Abstract

Retaining extremely polar metformin and its combination with other non-polar antidiabetic drugs can be difficult and challenging by using the most common reverse-phase high-performance liquid chromatography (RP-HPLC). The retention time of metformin is comparatively short when separated, utilizing C18-based RPC. Peak fronting and tailing effects are commonly observed with metformin (MET), its combination with highly polar antidiabetic medicines, and early elution with void volume. However, normal phase chromatography may occasionally be rendered ineffective due to the low solubility of polar analytes in organic solvents. Therefore, a different and complementary strategy is required for the separation of such a highly polar drug. A more advantageous platform is mixed-mode chromatography. The use of this newer technique of separations has been increased because of its advantages over C18-based RPC for its efficiency, selectivity, sensitivity, specificity and reproducibility towards simultaneous quantification of selected drugs. In this study, we compared two established chromatographic methods for simultaneous quantification of metformin and other moderately polar antidiabetic medications, including empagliflozin, remogliflozin, glimepride, and gliclazide, both in their pure form and in combination tablet dosage. An isocratic elution based on solvent a 15 mM ammonium acetate-Methanol: acetonitrile (10:90 v/v) was used as the mobile phase in first method, which is reversed-phase HPLC with a Zodiac C18 column and chromatographic separation on a 150 × 4.6 mm acclaimed mix mode HILIC-1 column with a particle size of 5 μm. In accordance with ICH standards, both approaches underwent exhaustive validation for linearity, accuracy, precision, selectivity, and resilience. Metformin, empagliflozin, ramogliflozin, glimepride, and gliclazide have not yet been reported to be separated using any method.