Bioaffinity ultrafiltration coupled with HPLC-ESI-MS/MS for screening potential α-glucosidase inhibitors from pomegranate peel

摘要

Pomegranate peel (PoP) contains plenty of bioactive compounds and exhibits strong activity to prevent postprandial hyperglycaemia and improve diabetes mellitus. Presently, bioaffinity ultrafiltration coupled with high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS/MS) is employed to screen and identify the efficient alpha-glucosidase inhibitors in PoP and the detailed inhibitory mechanisms are further investigated. The results show that many substances, including ellagic acid, kaempferol, gallic acid, and resveratrol in PoP reveal strong activity to inhibit alpha-glucosidase and ellagic acid (EA) is screened as the most effective compound. Further research indicates that EA plays a competitive and reversible inhibition role against alpha-glucosidase with the value of Ki was 6.24 x 10(5) mol/L. EA also directly interacts with the amino acids of alpha-glucosidase mainly via van der Waals forces and hydrogen bonds, thereby, influencing the secondary structure and stability of alpha-glucosidase. Finally, the alpha-glucosidase inhibitory activity of EA is further confirmed to significantly reduce postprandial blood glucose in vivo.

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