射干中乳酸脱氢酶抑制剂的筛选分离与质谱分析

摘要

本研究采用高效液相色谱-电喷雾质谱联用技术(HPLC-ESI-MS)对射干提取物中异黄酮类化学成分进行分析鉴定,以乳酸脱氢酶作为生物靶分子,运用超滤质谱技术筛选出6种酶抑制剂,分别为鸢尾苷(tectoridin)、鸢尾新甘A(IristectorinA)、野鸢尾苷(iridin)、鸢尾苷元(tectorigenin)、野鸢尾黄素(irigenin)和次野鸢尾黄素(irisflorentin),并采用半制备型高效液相色谱技术(Semi-prep·HPLC)分离纯化射干提取物中的活性成分.所得单体分离收集液经分析型高效液相色谱法检测,纯度均大于90％.结果证明利用液相色谱-超滤-质谱-半制备型高效液相色谱联用技术可以快速筛选、鉴定、分离射干中的活性物质.此方法对于筛选酶抑制剂有快速和灵敏等优势,具有广泛的应用前景.%In this study,high performance liquid chromatography coupled with photo-diode array detector and electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to screen and identify isoflavonoids with lactate dehydrogenase inhibitory activity from Belamcanda chinensis extracts.Lactate dehydrogenase was used as biotargeting molecule,six major compounds in B.chinensis was identified as lactate dehydrogenase inhibitors using ultrafiltration methods,namely tectoridin,iristectorin A,iridin,tectorigenin,irigenin,and irisflorentin.Following this,semi-preparative high performance liquid chromatography(semi-prep · HPLC) was applied to separate and isolate the active constituents.These compounds were then isolated by semi-prep HPLC with purity higher than 90％.Our results demonstrated that systematic isolation of bioactive components from B.ehinensis guided by ultrafiltration HPLC-ESI-MS was a feasible and efficient technique.It can be extended for the identification and isolation of other enzyme inhibitors.