Development and validation of rp-hplc method for the simultaneous estimation of linagliptin, empagliflozin and metformin in solid dosage forms

Abstract

The purpose of the investigation was to develop a simple, rapid and accurate RP-HPLC method to determine assay of Linagliptin, Empagliflozin and Metformin in solid dosage forms. The chromatographic separation was performed on Kromosil 250 x 4.6 mm, 5μm. Eluents were monitored on PDA detector at a wavelength of 233 nm using a Buffer: Acetonitrile (45:55v/v). The column temperature was maintained at 30°C. Validation parameters such as system suitability, linearity, precision, accuracy, specificity, limit of detection (LOD), limit of quantification (LOQ), Stability of sample and standard stock solutions and robustness were studied as reported in the ICH guidelines. The retention time for Linagliptin, Empagliflozin and Metformin was 2.370min, 2.787min and 3.419 min respectively. Assay method further evaluated for Linagliptin, Empagliflozin and Metformin analysis at low concentration of analyte and found limit of detection is 0.02 μg/ml, 0.08 μg/ml and 9.22μg/mlrespectively and limit of Quantitation is 0.05μg/ml, 0.25μg/mland 27.94μg/mlrespectively. The percentage recovery of Linagliptin, Empagliflozin and Metformin was 99.99%, 99.47% and 100.01% respectively. The %RSD for Linagliptin, Empagliflozin and Metformin was 0.5%, 0.7% and 0.2%respectively. Linearity of Linagliptin, Empagliflozin and Metformin performed from 25% to150% and the R2 is 0.999, intercept and slope found to be y = 25605x + 1357, y = 40970x + 3932 and y = 1497.x + 12010 respectively. The method was fast, accurate, precise and sensitive hence it can be employed for routine quality control of Linagliptin, Empagliflozin and Metformin containing drug in quality control laboratories and pharmaceutical industries.