Novel Stability Indicating Lc-MS/MS Method for the Simultaneous Estimation of Remogliflozin Etabonate and Vildagliptin Human Plasma

Abstract

The present study intended to develop an easy and novel Liquid Chromatography–Mass Spectrometry/Mass spectrometry (LC–MS/MS) method for simultaneous assay of remogliflozin etabonate and vildagliptin, a combined formulation used in treatment of type II diabetes in human plasma. Alogliptin drug was selected as internal standard and the analytes were isolated from the spiked plasma matrix using liquid-liquid extraction procedure and the extracts were chromatographed on Inertsil ODS (4.6 mm×100 mm, 5 μm) C18 column. The mobile phase comprises of methanol, acetonitrile and 0.1 % formic acid in 40:50:10 (v/v) at 0.5 mL/min flow rate and analysis was completed within 6 min run time. The method produces peaks with acceptable symmetry and resolution with acceptable system suitability at 2.6 min for remogliflozin etabonate, 2.7 min for vildagliptin, 1.2 min for alogliptin (internal standard). Multiple Reaction Monitoring (MRM) mode was used for the mass spectral characterization of column eluents using mass detector. In the mass spectral studies confirms the characteristic fragment ion transitions at m/z of 523 to m/z of 247 as MH+ ion for remogliflozin, m/z of 304 to m/z of 180 as MH+ ion for vildagliptin. The method can detect both the analyte up to 1.5 ng/mL and having lower limit of quantification (LLOQ) of 5 ng/mL. The method having broad calibration range of LLOQ to 300 ng/mL and was validated for precision, accuracy, stability studies such as freeze-thaw, short term and long-term stability in LLOQ, MQC (medium quality control concentration) and HQC (high quality control concentration) levels and produce acceptable results. Hence it can be confirmed that the method can be adopted for assay of remogliflozin etabonate and vildagliptin simultaneously in plasma samples.