Interest in the antioxidant and antidiabetic activity of natural products are growing vastly in the modern world. Thin layer chromatography-bioautography-mass spectroscopy (TLC-bioautography-MS) plays an important role in chemico-biological screening of natural sources. TLC combined with 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) free radical, -amylase and -glucosidase bioassay were used to evaluate antioxidant and antidiabetic activities, respectively, in different extracts of (Hanzal), a well-known traditional Indian Unani medicinal plant. To develop a TLC-bioautographic-MS method for DPPH, -amylase, and glucosidase inhibitors in different extract of fruits. Fruits of were successively extracted with toluene, dichloromethane, ethyl acetate, methanol, and water. TLC solvents were developed, and bioautographic-MS analysis was carried out to identify the antioxidant and antidiabetic compounds. H maximum in extracts of at 254 and 366 nm, respectively, whereas in toluene extract at 254 and 366 nm. Based on TLC-bioautography-MS, maximum number of extracts of , all the extracts have shown antidiabetic activity. On the other hand, there were no antioxidant compounds in methanolic extract of . The results of this study reveal that TLC-bioautography-MS-guided strategy used to identify antioxidant and antidiabetic compounds of is very useful technique for high-throughput screening of bioactive compounds. TLC-MS bioautography is a simple and fast to enables bioactive compounds present in extracts.