Bisma Jan^{1, 2}, Sultan Zahiruddin², Parakh Basist², Mohammad Umar Khan^{1, 2}, Sageer Abass³, Rikeshwer Prasad Dewangan⁴, Sayeed Ahmad^2,

Morus alba (M. alba) popularly known as white mulberry is a well-known and globally utilized functional food and nutraceutical. The effect of different brewing time (BT) on antioxidant and antihyperglycemic compounds released from M. alba leaves infusion was studied. Major compounds responsible for antioxidant activity were identified and quantified by high-performance thin-layer chromatography (HPTLC). Additionally, thin layer chromatography-mass spectrometry-bioautography (TLC-MS-bioautography) analysis was performed to identify the compounds responsible for antioxidant and antihyperglycemic activity. In vitro α-amylase and α-glucosidase inhibitory activity grew in a linear manner with each BTs. Based on the findings, 7 and 10 minutes of BTs at 98°C were chosen as optimal for infusion preparation. Major phenolic acid i.e. chlorogenic acid at 7 and10 minutes of BT in M. alba infusion was quantified as 89.93 µg/mL and 95.0 µg/mL respectively. However, major flavonoid rutin was calculated as 5.56 µg/mL and 8.79 µg/mL respectively. Deoxynojirimycin (DNJ), which is a potent α-glucosidase inhibitor was identified in M. alba infusion by ultra performance liquid chromatography-mass spectrometry (UPLC-MS) at Rt of 0.88 minutes. TLC-MS-bioautography revealed that chlorogenic acid, rutin, and DNJ are predominant antioxidant and antihyperglycemic compounds in M. alba infusion. UPLC-MS fingerprinting analysis concluded that M. alba infusion at 98°C for 7 and 10 minutes may be a good source of important alkaloids (fagomine), flavonoids (quercetin, morin), and carboxylic acid (fumaric acid), and other metabolites responsible for its antioxidant and antihyperglycemic activity favoring its utilization as a functional drink with therapeutic potential.

Morus alba, DNJ, functional drink, antihyperglycemic, antioxidant, TLC-MS bioautography, UPLC-MS