Diabetes mellitus, which has high rates of disability and mortality, is one of the main causes of public health concerns worldwide. It currently poses serious medical and societal issues. Different bio-analytical techniques have been developed to identify the pharmaceutical formulation of the anti-diabetic drug. Due to the continual requirement to achieve improved sensitivity, accuracy, and speed of analysis in complex biological samples, the development of bio-analytical sample preparation techniques has grown more difficult over time (e.g., blood, serum, plasma, saliva, feces, and urine). This is a mini review, and its goal is to analyze how LC-MS/MS has been used to for the pharmaceutical formulation of the anti-diabetic drugs. The method was validated for linearity, accuracy, precision, specificity, selectivity, and stability.