UHPLC/MS Identifying Potent α-glucosidase Inhibitors of Grape Pomace via Enzyme Immobilized Method.

α-Glucosidases have been a major target in controlling and managing postprandial blood glucose and therefore diabetes treatment. This study aims to further identify and purify active compounds from the most active ethyl acetate fraction collected previously in Tinta Cão grape pomace extract (TCEE) using a newly developed and highly effective immobilization method, including obtaining compounds previously shown to inhibit the enzyme. Purification used crosslinked chitosan beads with α-glucosidases bound to polymer, which acted as immobilized enzyme vehicle to collect inhibitors. Compounds absorbed into the beads were eluded using methanol, where collected fraction was subjected to UHPLC-MS analysis to identify active compounds. Results presented 5 major compounds: viniferifuran (amurensin H), p-coumaroyl-6-O-D-glucopyranoside, p-coumaroyl-6-O-hexoside, (epi)catechin-hexoside, 10-carboxyl-pyranopeonidin 3-O-(6''-O-p-coumaroyl)-glucoside. These findings indicated the particular molecules can be utilized as potent α-glucosidases inhibitors, and may be further tested for postprandial glucose control. PRACTICAL APPLICATION: A potential approach enriched and identified α-glucosidase inhibitors of grape pomace. Set-up of UHPLC/MS detection and identification of active compounds provide qualify assessment in developing grape pomace extract into potent dietary supplement and new drug for diabetes.