Development and Validation of an RP-HPLC method for Simultaneous Estimation of Empagliflozin, Linagliptin and Metformin in bulk and Combined Pharmaceutical Dosage Forms

Abstract

Antidiabetic agents used for the treatment of type 2 diabetes mellitus, such as Empagliflozin, Linagliptin and Metformin are often used in combination therapy. The objective of this study was to develop and validate a simple, reliable and reproducible RP-HPLC method for simultaneous determination of Empagliflozin, Linagliptin and Metformin in tablet formulation. The separation by chromatography took place by using 0.1% glacial acetic acid and acetonitrile at the ratio of 55:45 (v/v) and detecting by wavelength 222 nm. Total analytical time was 12 min and the retention times were 1.967 min for Metformin, 4.477 min for Linagliptin and 7.40 min for Empagliflozin. The method exhibited good linearity over Metformin, Linagliptin and Empagliflozin concentration ranges from 300-800µg/mL, 1.5-4µg/mL and 3-8µg/mL respectively. R2 values were 0.998 for Metformin, 0.997 for Linagliptin and 0.994 for Empagliflozin. Percent recovery ranged from 99-102% and the %RSD was in the range of 0.01-0.06% for precision assessments. Empagliflozin, Linagliptin and Metformin had limits of detection and quantification of 0.375 µg/mL and 1.235 µg/mL, and 3.75 µg/mL and 12.3 µg/mL, respectively. Validation was conducted following the ICH guideline and the validation parameters investigated were satisfactory. The proposed method can provide reasonably good separation and reliable quantitation of all three drugs in a 12-minute run time and can be used for systematic QC testing of the drugs together in their pharmaceutical dosage forms.