Exploration and Validation of an Advanced Reverse-Phase HPLC Approach for the Concurrent Analysis of Empagliflozin, Linagliptin, and Metformin in Pharmaceutical Dosage Forms

Abstract

A straightforward, precise, and reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed and validated for the simultaneous quantification of EMPA, LINA, and MET in pharmaceutical dosage forms. Chromatographic separation was conducted using an Eclipse XDB C₁₈ Column (250 mm × 4.6 mm i.d., particle size 5 \(\mu\)m, maintained at ambient temperature), with a mobile phase consisting of 0.01 N KH₂PO₄ and acetonitrile in a 50:50 v/v ratio. A pH 5.3 buffer, adjusted with o-phosphoric acid, was utilized. UV detection was carried out at 220 nm, and the retention times were 2.767, 2.362, and 3.276 minutes for EMPA, LINA, and MET, respectively. Calibration plots demonstrated linearity (r² > 0.999) over the concentration range of 2.5-15 \(\mu\)g/mL for EMPA, 1.25-7.5 \(\mu\)g/mL for LINA, and 250-1500 \(\mu\)g/mL for MET. The method underwent rigorous validation for accuracy, precision, specificity, linearity, and sensitivity. Successfully applied to the quantitative analysis of pharmaceutical dosage forms, the method exhibited no interference from any component of the pharmaceutical dosage form.

Validation studies affirmed the specificity, rapidity, reliability, and reproducibility of the method. High recovery and low relative standard deviation further affirmed the method's suitability for the routine determination of EMPA, LINA, and MET in pharmaceutical dosage forms.