Eco-Friendly LC-MS/MS Method for Quantification of Dapagliflozin and Linagliptin in Combined Dosage Form: Development, Validation and AGREE Assessment

Abstract

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder that requires effective therapeutic monitoring. Dapagliflozin and linagliptin, two commonly prescribed antidiabetic agents, act via distinct mechanisms—dapagliflozin inhibits sodium-glucose co-transporter- 2 (SGLT-2), while linagliptin is a dipeptidyl peptidase-4 (DPP-4) inhibitor. This study aimed to develop and validate a sensitive, precise, and green LC-MS/MS method for the simultaneous quantification of dapagliflozin and linagliptin in pharmaceutical dosage forms. Chromatographic separation was achieved on a Phenomenex Gemini C18 column (50 mm × 4.6 mm, 5 μm) using an isocratic mobile phase of methanol and 0.1% formic acid (80:20, v/v) at a flow rate of 0.5 mL/min. Detection was carried out using a mass spectrometer equipped with electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode, monitoring transitions at m/z 426.3→135.1 for dapagliflozin and 473.3→420.3 for linagliptin. The method exhibited excellent linearity over the range of 5–500 ng/mL for dapagliflozin (r² ≥ 0.9996) and 10–1000 ng/mL for linagliptin (r² > 0.9993). It demonstrated acceptable precision, accuracy, and stability in accordance with regulatory guidelines. Additionally, the AGREE metric yielded a score of 0.76, indicating good compliance with green analytical chemistry principles. The developed LC-MS/MS method is reliable, eco-friendly, and suitable for routine quality control of dapagliflozin and linagliptin in combined formulations. Its robustness also supports potential future applications in pharmacokinetic and bioequivalence studies.