α-Glucosidase is a key enzyme target for diabetes and obesity treatment.Inhibiting its activity can produce therapeutic effects.To efficiently discover natural product-derived α-glucosidase inhibitors, this study employed a strategy combining magnetic nanoparticles with α-glucosidase to selectively capture potential bioactive compounds, followed by rapid identification of captured components using ultra-high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS).Single-factor experiments determined optimal immobilization conditions:15% glutaraldehyde concentration, 7 U/mL α-glucosidase concentration, and 4 h immobilization time, achieving maximum relative enzyme activity.This optimized immobilized enzyme system was used to screen and identify 14 compounds from Cyclocarya paliurus extracts, the IC50 values of 10 active compounds and acarbose are as follows:isochlorogenic acid A(870.3 μg/mL), myricetin-3-O-β-D-glucoside(123.9 μg/mL), quercetin-3-O-glucuronide(587.2 μg/mL), quercitrin(320.9 μg/mL), myricetin(25.7 μg/mL), hyperoside(394.1 μg/mL), afzelin(272.7 μg/mL), kaempferol-3-O-β-D-glucuronopyranoside(524.9 μg/mL), kaempferol(21.6 μg/mL), quercetin(31.0 μg/mL), acarbose(0.62 μg/mL).The results demonstrate that immobilized enzyme technology holds strong potential for efficiently discovering therapeutic compounds in medicinal plants.
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