中药活性成分的快速分离分析一直是中药研究的重点和热点问题.该文建立了在线亲和固相萃取-高效液相色谱-二极管阵列检测-四极杆飞行时间质谱技术快速筛选中药中与α-葡萄糖苷酶有结合作用的活性成分的方法.四通阀和六通进样阀组成接口界面,收集亲和固相萃取柱中与α-葡萄糖苷酶有结合作用的活性成分,接着进入液相色谱系统进行分析鉴定.阳性对照品((+)-儿茶素)和阴性对照品(水杨酸)混合物筛选确定了方法的特异性和准确性.接着,以玉竹为研究对象,筛选出9种主要的α-葡萄糖苷酶抑制活性化合物,包括5种苯乙基肉桂酰胺类化合物,4种双氢高异黄酮类化合物.结果表明建立的方法简便、快速、特异性强,可用于任何复杂体系中α-葡萄糖苷酶结合活性成分的筛选.
Screening and analysis of bioactive compounds from natural products is a challen-ging work due to their complexity.This paper reports the hyphenation of affinity solid-phase extraction column (ASEC) using immobilized α-glucosidase and high performance liquid chromatography-diode array detector-quadrupole time-of-flight mass spectrometry (HPLC-DAD-Q-TOF-MS) for screening and identification of α-glucosidase binding compounds from Chinese medicines.Affinity solid-phase extraction (ASE) system was hyphenated with HPLC system via a four-port switching valve and a six-port injection valve as an interface for transferring effluents from ASE column to HPLC.Positive control ((+)-catechin) and negative control (salicylic acid) were performed with the approach to verify its specificity and reproducibility.Subsequently, the developed system was applied to the screening and identification of α-glucosidase inhibitors from Polygonatum odoratum.Five phenethyl cinnamides (N-cis-feruloyloctopamine, N-trans-p-coumaroyloctopamine, N-trans-feruloyloctopamine, N-trans-p-coumaroyltyramine and N-trans-feruloyltyramine) and four homoisoflavanones ((3R)-5,7-dihydroxyl-3-(2′,4′-dihydroxylbenzyl)-chroman-4-one, (3R)-5,7-dihydroxyl-6-methyl-3-(4′-hydroxylbenzyl)-chroman-4-one, (3R)-5,7-dihydroxyl-6-methyl-8-methoxyl-3-(4′-hydroxylbenzyl)-chroman-4-one and (3R)-5,7-dihydroxyl-6,8-dimethyl-3-(4′-hydroxylbenzyl)-chroman-4-one) with α-glucosidase inhibitory activities were identified.The results were in accordance with those of ultrafiltration screening.With the online system developed, here we present a feasible, selective and effective strategy for the rapid screening and identification of enzyme inhibitors from complex mixtures.
参考文献
| [1] | Wang, Y;Kong, L;Lei, XY;Hu, LH;Zou, HF;Welbeck, E;Bligh, SWA;Wang, ZT.Comprehensive two-dimensional high-performance liquid chromatography system with immobilized liposome chromatography column and reversed-phase column for separation of complex traditional Chinese medicine Longdan Xiegan Decoction[J].Journal of Chromatography. A,200911(11):2185-2191. |
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