磁珠富集与液相色谱-质谱联用结合筛选枳壳中脂肪酶抑制剂

Rapid Screening of Lipase Inhibitors from Fructus aurantii Using Lipase-immobilized Magnetic Beads Coupled with High-performance Liquid Chromatography-Mass Spectrometry

提出了一种磁珠收集与液相色谱-质谱联用(LC-MS)结合用于快速筛选中药提取物中的脂肪酶抑制剂的方法.通过制备键合脂肪酶的磁珠,将其与枳壳总黄酮孵育后,筛选出枳壳中脂肪酶的配体.通过LC-MS分析发现,4个化合物均是潜在的脂肪酶抑制剂,鉴定其分别为柚皮苷、新橙皮苷、橙皮苷和枸橘苷.对4个化合物进行了体外脂肪酶抑制活性验证.研究结果表明,新橙皮苷、橙皮苷和枸橘苷具有显著的脂肪酶抑制活性,IC50分别为48.04,52.45和46.18μg/mL,其中枸橘苷具有脂肪酶抑制活性为首次报道.结果表明,磁珠富集与LC-MS集成技术能够用于快速发现中药活性成分.

A new method for the rapidly screening of lipase inhibitors from extracts of traditional Chinese medicine using lipase-immobilized magnetic beads coupled with high-performance liquid chromatography-mass spectrometry（LC-MS） was proposed. After the lipase-immobilized magnetic beads were prepared, they were incubated with total flavonoid of Fructus aurantii to fish the ligands of lipase. Then, the eluent of magnetic beads＇ screening was analyzed by LC-MS. Four compounds, as naringin, hesperidin, neohesperidin and poncirin, were identified as the potential lipase inhibitors. Furthermore, the in vitro inhibitory activities against lipase of four compounds were tested. The results showed that hesperidin, neohesperidin and poncirin exhibit lipase inhibitory activities, while poncirin was firstly reported to be a lipase inhibitor. Our research showed that active compounds could be rapidly screened from extracts of traditional Chinese medicine via lipase-immobilized magnetic beads coupled with LC-MS.