Direct Screening of α-Glucosidase Inhibitor from Natural Plant Extracts by Immobilized Enzymes Affinity Chromatography Fishing Combined with UPLC-Q-TOF-MS Analysis
A novel enzyme inhibitor screening method was established by a combination of immobilization of α-glucosidase(AGH) on agarose beads and ultra performance liquid chromatography-quadrapole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) for fishing and identification of active components directly from natural plant extracts.The AGH was first immobilized by reaction with the activated agarose and then incubated with the natural plant extracts for ligand fishing.Three components,(-)-epigallocatechin-3-gallate(EGCG),(-)-gallocatechin -3-gallate(GCG) and (-)-epicatechin-3-gallate(ECG),in the extract of leaves of Camellia sinensis (green tea) were fished out by their specific affinity binding between the ligands and the enzyme protein and then identified by UPLC-Q-TOF-MS.Furthermore,the above three components isolated from the green tea extracts were tested by traditional enzyme inhibition assay,the results showed each of these compounds was a potent α-glucosidase inhibitors (IC50<5 μM).This work demonstrates that this new immobilized enzyme-UPLC-MS method may provide an alternative for rapid,effective and direct screening enzyme inhibitors from natural plant extracts.
Shiren Deng Honghin Xiao
National Chromatographic R&A Center, Dalian Institute of Chemical Physics, the Chinese Academy of Sc National Chromatographic R&A Center, Dalian Institute of Chemical Physics, the Chinese Academy of Sc
国内会议
海口
英文
304-304
2012-11-01（万方平台首次上网日期，不代表论文的发表时间）