黄连原小檗碱类生物碱快速制备及清除超氧阴离子活性研究

Study on rapid preparation of protoberberine alkaloids from Coptidis rhizoma and their superoxide anion scavenging activities

为建立两步法快速制备黄连原小檗碱类生物碱,并研究其体外清除超氧阴离子的活性,通过制备薄层色谱法分离黄连95%乙醇提取物,半制备型高效液相色谱纯化,快速得到6个主要的原小檗碱类生物碱。再以黄嘌呤-黄嘌呤氧化酶反应体系产生超氧阴离子,WST-1作为超氧阴离子的探针,采用微孔板法评价黄连原小檗碱类生物碱对超氧阴离子的体外清除活性。结果表明:(1)两步法操作简单,分离快速,所得单体纯度高。(2)非洲防己碱、药根碱和表小檗碱对体外超氧阴离子有一定程度的清除活性且呈浓度依赖关系,具有一定的抗氧化作用。这一研究为黄连的深入研究提供了一定的理论依据。

The present study aimed to establish a two-step method for the rapid preparation of protoberberine alkaloids from Coptidis rhizoma and evaluate their in vitro superoxide anion scavenging activities. The 95% ethanol extract of C.rhizoma was separated by preparative thin layer chromatography and then purified by semi-preparative HPLC to obtain six major protoberberine alkaloids. Superoxide anion was produced by the reaction system of xanthine and xanthine oxidase. WST-1 was used as a probe to detect superoxide anion. The in vitro scavenging activities of the protoberberine alkaloids from C.rhizoma on superoxide anion were evaluated by a platereader-based microassay. The results indicated that:(1) The two-step method had the advantages of simple operation, rapid separation and high purity of isolated monomer compounds.(2) Columbamine, jatrorrhizine and epiberberine showed in vitro superoxide anion scavenging activities to some extent and concentration dependent relationships were observed, which indicated that columbamine, jatrorrhizine and epiberberine had certain anti-oxidative effects. This study provides a theoretical basis for the further study of C.rhizoma.