RationaleThe present study aims to establish structures of the degradation products of an anti‐diabetic drug, Imeglimin (IMG) approved for the treatment of type 2 diabetes mellitus in the year 2021. Degradation pathways are proposed along with in silico toxicity assessments of the observed degradation products (DPs) of the drug.MethodsA reversed‐phase high‐performance liquid chromatography (RP‐HPLC), equipped with a photodiode array detector, was used to separate the observed DPs with a Phenomenex Luna PFP (250 × 4.6 mm, 5 μm) column, using 10 mM ammonium formate (pH 4.5) and methanol as mobile phase. Liquid chromatography quadrupole time of flight mass spectrometry (LC‐Q‐ToF‐MS/MS) and nuclear magnetic resonance (NMR) spectroscopy were employed for structural elucidation. Zeneth and Derek suites were used for in silico assessments.ResultsA total of four degradation products were observed, which were successfully separated on an RP‐HPLC. The structural characterization of three of the four DPs was achieved using LC‐Q‐TOF‐MS/MS by employing electro spray ionization as well as atmospheric pressure chemical ionization. Additionally, DP‐3 was isolated using a preparative HPLC and was characterized by NMR. Computationally predicted structures were compared with the experimental observations.ConclusionAn HPLC method, capable of separating the Imeglimin and its four DPs, was developed and validated as per the ICH Q2(R1) guideline. Structure elucidation reveals a variety of products with metformin as one of the identified DPs along with a metabolite. The toxicity potential of DPs was assessed through docking studies.