Abstract
Objective: The proposed method aims to develop a simple, rapid, sensitive and validated isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of linagliptin and empagliflozin in human plasma.Methods: Chromatography was performed on waters 2695 HPLC equipped with a quaternary pump. The separation was carried using discovery C18 (250×4.6×5) column, buffer: acetonitrile (68:32) as mobile phase with 1 ml/min flow rate. The analyte detection was monitored at 218 nm.Results: Retention time of linagliptin, empagliflozin and internal standard was found at 6.421, 4.696, and 4.074 min respectively. The peaks were found to be free of interference. The method is validated over a dynamic linear range of 0.01-10.0 µg/ml for both drugs with a correlation coefficient of 0.998. The precision and accuracy of samples of six replicate measurements at lower limit of quantification (LLOQ) level were within the limit. The analytes were found to be stable in human plasma at-28 °C for 37 d.Conclusion: The stability, sensitivity, specificity and reproducibility of this method make it suitable for the determination of linagliptin and empagliflozin in human plasma.
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