Comprehensive Profiling And Quantification Of Novel Antidiabetic Pharmaceuticals (Dapagliflozin And Saxagliptin): An Integrated Approach Utilizing Liquid Chromatography-Mass Spectrometry (LC-MS) For Structural Elucidation, Metabolic Pathway Identification, And Bioavailability Assessment

Abstract

Background: Diabetes mellitus is a global health challenge, necessitating the development of novel antidiabetic therapies. Dapagliflozin, a sodium-glucose cotransporter-2 (SGLT-2) inhibitor, and Saxagliptin, a dipeptidyl peptidase-4 (DPP-4) inhibitor, represent critical advancements in diabetes management. Comprehensive profiling of their structures, metabolic pathways, and bioavailability is essential for optimizing therapeutic efficacy and safety. Objectives:  This study aimed to:  Elucidate the structures of Dapagliflozin and Saxagliptin through MS/MS analysis.  Identify and characterize their metabolites.  Assess their pharmacokinetics and bioavailability using LC-MS techniques. Methods  Instrumentation: LC-MS was utilized with electrospray ionization (ESI) and multiple reaction monitoring (MRM) for precise detection.  Sample Preparation: Plasma and urine samples were processed through protein precipitation and solid-phase extraction.  Metabolic Profiling: In vitro assays with liver microsomes and in vivo pharmacokinetic studies were conducted.  Bioavailability: Plasma concentration-time data were analyzed, and pharmacokinetic parameters such as C_max, T_max, and AUC were calculated. Results Structural Elucidation:  Dapagliflozin exhibited key fragments at m/z 408 and 345, confirming its glycosidic and aromatic features.  Saxagliptin displayed fragmentation at m/z 409, 215, and 150, identifying its pyrrolidine and amide groups. Metabolic Pathways:  Dapagliflozin underwent hydroxylation and glucuronidation, yielding active metabolites.  Saxagliptin primarily underwent hydroxylation and lactam cleavage. Bioavailability:  Dapagliflozin: C_max = 6.2 µg/mL at T_max = 4 hours; AUC (0–24) indicated moderate bioavailability.  Saxagliptin: C_max = 7.0 µg/mL at T_max = 4 hours; higher bioavailability compared to Dapagliflozin. Conclusion LC-MS proved instrumental in characterizing the structures, metabolites, and pharmacokinetic profiles of Dapagliflozin and Saxagliptin. These findings provide critical insights into their ADME properties, supporting their clinical use in diabetes management. Future studies should explore drug-drug interactions and individual metabolic variations to enhance therapeutic outcomes.