A SIMPLE, SENSITIVE AND FAST SINGLE STEP EXTRACTION METHOD FOR DETERMINATION OF EMPAGLIFLOZIN IN HUMAN PLASMA USING LC-MS/MS

摘要

A simple, sensitive, and fast single-step extraction method for estimating empagliflozin in human plasma using LC-MS/MS was developed and validated for pharmacokinetics evaluations. Plasma samples were basified before solid-phase extraction on SOLA (30 mg / 1 mL cartridges). Separations were carried out on a normal reverse phase C18 column (Hypersil BDS 100 × 4.6, 5μ mm column) for 3.5 minutes at a flow rate of 0.6 mL/min. Ten μL of the SPE eluent is directly injected onto LC-MS/MS to quantify the analyte from 1.563-800.000 ng/mL using a single SRM transition (m/z: 449.140à371.100) in negative ion mode. During method validation, selectivity, matrix effect, recovery, carry-over effect, stability studies, inter-day, and intra-day precision and accuracy experiments were conducted per USFDA guidelines. Method validation data has successfully met the acceptance criteria making it suitable for use in routine bio-analytical laboratories. The scope of this assay can be extended to cover the requirement of preclinical, toxicology, and PK/PD studies.