Development and bioanalytical validation of a high-sensitivity LC-MS/MS assay for quantitative determination of sotagliflozin in rabbit plasma and its stability across controlled storage conditions

Sotagliflozin is a dual SGLT-1 and SGLT-2 inhibitor approved by the FDA in 2023 which has emerged as a novel therapeutic agent for the management of type 2 diabetes mellitus. In this study robust and sensitive LC-MS/MS method was developed and validated for quantification of sotagliflozin in rabbit plasma as rabbit is commonly used non-rodent model for preclinical research. Sample preparation involved protein precipitation for efficient analyte extraction from rabbit plasma. Chromatographic separation was performed utilizing on BDS Hypersil C18 column (100 mm x 4.6 mm, 5 μm) using mobile phase composed of methanol (85%) and 5 mM ammonium acetate in milli-Q water (15%) and rolipram as internal standard. The method employed flow rate of 0.7 mL/min with a total runtime of 4 min and an injection volume of 10 µL. Method validation was carried out in accordance with ICH M10 guidelines, covering precision, accuracy, selectivity, recovery, and stability at different storage conditions. The method was found to be linear over the concentration range of 10-1280 ng/mL with sensitivity of 10.20 ng/mL (LLOQ) in rabbit plasma. Recovery of analyte from the rabbit plasma was found to be > 92% with stability > 99% at different storage conditions (viz., room temperature, autosampler, freeze-thaw and frozen). Overall, the developed LC-MS/MS method offers a simple, precise and reproducible approach for the quantification of sotagliflozin in rabbit plasma and is well-suited for application in pharmacokinetic and other preclinical studies.