Objective: Herbal medicines have long been used effectively in the treatment of Diabetes Mellitus. Cardiospermum halicacabum(C. halicacabum) has been used for several centuries for the treatment of various ailments. The present work was undertaken to study the effect of isolated bioactive compounds from leaf of C. halicacabum on glucose uptake in L-6 cell lines.Methods: The hydro alcoholic leaf extract of C. halicacabum was subjected to preliminary phytochemical screening and isolated eleven compounds were tested for cytotoxicity by MTT assay. Compound 1 and 8 were selected for further glucose uptake assay based on cytotoxicity concentration CTC<inf>50</inf> value. Structure of compound 1 and 8 was identified by NMR technique.Results: Phytochemical analysis showed the presence of alkaloids, flavonoids, tannins and terpenoids. Compound 1 showed moderate cytotoxic activity to L6 cell line when compared to compound 8. Compounds 1 and 8 were identified as Kaempferol-3-o-α-l-rhamnoside and Apigenin-7-o-β-d-glucuronide. In in vitro glucose uptake assay Kaempferol-3-o-α-l-rhamnoside showed increase in glucose uptake by 11.38 ± 2.04% and Apigenin-7-o-β-d-glucuronide by 15.97 ± 2.16% over control.Conclusion: Kaempferol-3-o-α-l-rhamnoside and Apigenin-7-o-β-d-glucuronide isolated from C. halicacabum leaf extract enhance the glucose uptake significantly. They exhibit hypoglycemic activity and stimulate glucose uptake in L-6 skeletal muscle cells. This study can bring a promising role for this plant in the management of Diabetes mellitus. © 2015 Elsevier B.V., All rights reserved.