Since the beginning of the medicinal plants have been used to treat a variety of illness. A systemic disorders called Diabetes mellitus is characterized by hyperinsulinemia, significantly increased blood lipid and fat concentrations and hyperglycaemia. The anti-diabetic activity of ethanolic crude extract of Pracecitrullus fistulous (P. fistulous) and its active fraction were evaluated by in-vitro assays. Extraction of P. fistulous leaves was carried out using ethanol and further, Ethyl acetate and n-butanol fractions were obtained. To investigate the hypothesis of hypoglycemic effects of ethanolic extract of P. fistulous (EEPF), Ethyl acetate fraction of P. fistulous (EAPF), n-butanol fraction of P. fistulous (NBPF) and Acarbose (Standard drug), (100- 100µg/ml) were used for the α-amylase and α-glucosidase enzyme inhibition study. EEPF, EAPF, NBPF were also studied for DPPH, Reducing power assay, Hydrogen peroxide scavenging assay, HPTLC. From the above the studies carried out the active fraction were subjected for the MIN-6β cell line study. The insulin secretagogue actions of EEPF, EAPF and NBPF were investigated utilizing a mouse insulinoma beta cell line (MIN6-β). The study has revealed that the EAPF showed the highest anti-diabetic potential in comparison to the EEPF and NBPF.