The scientific evaluation of the antidiabetic efficacy of aqueous extract of Trichosanthes dioica fruits on streptozotocin-induced diabetic rats is being presented. The graded doses of the extract, viz., 500, 750, 1,000, and 1,250 mg/kg body weight (bw), were administered orally, and it was observed that the blood glucose concentration decreased in a dose-dependent manner. The dose of 1,000 mg/kg bw showed the maximum fall of 23.8% and 19.1% in blood glucose level (BGL) during fasting BGL and glucose tolerance test (GTT) studies, respectively, of nondiabetic rats. Whereas in the case of subdiabetic and mild diabetic models, the same dose showed reduction in BGL of 22.0% and 31.4% during GTT. The study also involves the first use of laser-induced breakdown spectroscopy as a sensitive analytical tool to detect the elemental profile responsible for the antidiabetic activity of aqueous extract of T. dioica fruits that exhibits the antidiabetic activity. High intensities of Ca, Mg, and Fe indicate large concentrations of these elements in the extract, since according to Boltzmann’s distribution law, intensities are directly proportional to concentrations. The higher concentrations of these glycemic elements, viz. Ca, Mg, and Fe, are responsible for the antidiabetic potential of T. dioica as well as other plant already reported by our research group.