亲和超滤耦联液相色谱-质谱快速检测山楂叶中的α-淀粉酶抑制剂

Characterization of α-Amylase Binding Agents from Hawthorn Leaf Using Ultrafiltration and Liquid Chromatography Tandem Mass Spectrometry

建立了用于快速检测山楂叶中α-淀粉酶抑制剂的生物亲和超滤方法。考察了磷酸盐缓冲液的pH值、离子强度、孵育时间、孵育温度、α-淀粉酶浓度等影响亲和超滤的因素,最终选择了10 mmol/L磷酸盐缓冲液(pH 7.4)作为孵育溶液,在37℃孵育45 min,淀粉酶浓度为10 g/L。在优化条件下,将本法应用于山楂叶总黄酮的α-淀粉酶抑制剂的筛选,得到3个活性成分,分别鉴定为槲皮素-3-o-鼠李糖基-(1→4)-葡萄糖基-(1→4)-鼠李糖苷、牡荆素-2″-葡萄糖苷、牡荆素-2″-鼠李糖苷。这3个化合物的α-淀粉酶抑制活性为首次报道。

An affinity based ultrafihration method was established to rapid screening amylase inhibitors from hawthorn leaf. Various factors were optimised including the pH of the incubation buffer, ion strength, incubation time, incubation temperature and the concentration of a-amylase. Kaemp-ferol was chosen as a model compound. The condition chosen for screening was as follows： pH 7.4, ion strength 10 mmol/L, incubation time 45 min, incubation temperature 37 ℃, amylase concentration 10 g/L. Then, the established method was applied to screening a-amylase inhibitors from extract of hawthorn leaf and three active compounds were obtained.