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Diabetes mellitus is the most common metabolic disorder affecting more than 422 million people worldwide. The available pharmacological approaches for the prevention and treatment of diabetes are not adequate due to associated side effects and higher costs. For centuries, ancient cultures across the globe emphasized the critical role of natural remedies in managing the diabetes by using them. Asparagus gonoclados is a traditional folk remedy used for diabetes mellitus in India. The present investigation was taken up to evaluate the anti‐diabetic activity of the A. gonoclados (Liliaceae) root tubers. Different solvent extracts (aqueous, ethanol, ethyl acetate and hexane) were prepared from A. gonoclados root tubers and were tested for anti‐hyperglycemic activity in streptozotocin induced diabetic rats and, anti‐α‐glucosidase and anti‐α‐amylase activity in vitro . Among the different solvent extracts, oral administration of ethanol extract (AGEE) of A. gonoclados, at a dose 500 mg/kg b.w, has produced maximum (67%) reduction in fasting blood glucose levels in diabetic treated rats after 6 hrs when compared to the glibenclamide (20 mg/kg b.w.) treated group. AGEE also has revealed the dose‐dependent inhibitory effect on α‐glucosidase and α‐amylase activities that is comparable to α‐glucosidase inhibitor drug, Acarbose. Phytochemical analysis of AGEE revealed the presence of steroidal saponins, sapogenins, phenols, flavonoids, phytosterols, tannins, triterpenes, carbohydrates, alkaloids and proteins. AGEE has the richest quantity of total phenolics (138.4±0.39 μg/mg gallic acid equivalents), flavonoids (64.8±0.54 μg/mg quercetin equivalents) and steroidal saponins (12.9±0.11 μg/mg sarasapogenin eqivalents). Further characterization of ethanol extract was carried out by LC‐ESI‐MS/MS analysis and fifteen compounds were identified. Our studies are the first to report anti‐diabetic activity of A. gonoclados root tubers. We conclude that A. gonoclados has potential therapeutic importance in the management of diabetes mellitus. Support or Funding Information This study is funded by Indian Council of Medical Research (ICMR, India) through major research project (No.45/18/2012). This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .