目的:建立大黄黄连泻心汤中主要化学成分的HPLC-DAD分析方法,结合LC/MS/MS进行该复方不同药味配伍前后浸渍剂中化学成分的归属研究.方法:在HPLC分析的色谱条件中,采用Luna苯基柱(250mm×4.6mm,5μm),DAD检测器在紫外区200～400nm范围内扫描,流动相为乙腈-0.5%三乙胺(磷酸调节pH至3.5),梯度洗脱,流速为1mL·min-1;进样量为20μL.通过对大黄黄连泻心汤中各药味单独和不同药味配伍前后浸渍剂中各色谱峰的保留时间和吸收光谱与对照品进行比较,并结合LC/MS/MS进行相关色谱峰的鉴定和归属研究.结果:本研究建立的HPLC-DAD方法,可较好的进行大黄黄连泻心汤中主要色谱峰的分离和对应色谱峰光谱扫描;在HPLC图谱共标出了20个主要色谱峰,结合对照品对其中的黄连小檗碱等11个化学成分进行了鉴定和来源归属;此外,对某些化学成分进行HPLC分离和LC/MS/MS定性,推断出了黄连碱等4个成分,并判断了它们的归属.结论:本研究建立的HPLC-DAD方法准确、重复性好,可进行大黄黄连泻心汤浸渍剂中主要化学成分的定性分析;对15个色谱峰进行的定性和归属研究结果可对揭示该复方的物质基础、配伍规律和质量控制提供实验数据支持.

The aim of the study is to determine the major chemical components in different herbs and combinations of the Chinese medicine formula Dahuang Huanglian Xiexin Decoction (DHXD) which is composed of Rheum officinalis, Coptis sinensis, and Scutellaria baicalensis, with HPLC-DAD and LC/MS/MS methods. A HPLC-DAD method was established with the Luna ODS column (250mm×4.6mm，5μm). The mobile phase was acetonitrile - 0.5% triethylamine (adjusted by phosphoric acid, pH 3.5) gradient elution with the flow rate of 1.0 mL·min-1. The sample size was 20μL. The chromatographic peaks of individual herbs and the DHXD formula were identified by comparing the relative retention time and UV absorption spectrum with the reference compounds, together with LC/MS/MS. The HPLC-DAD method was shown to be accurate and reproducible. 20 chromatographic peaks were observed totally. 11 peaks including berberine from Coptis were identified using reference compounds, and 4 peaks including coptisine were determined by LC/MS/MS. The results showed that the HPLC-DAD method is suitable for qualitative analysis of major chemical components in DHXD. The identification of the 15 chemical components and determination of their source herbs in the formula provide new analytical evidence on the chemical basis and combinational principles, as well as quality control of DHXD.

国家自然科学基金面上项目(30873428):大黄黄连泻心汤主要化学成分分析与生理药动学研究，负责人：姚美村；广东省中医药管理局面上项目(2009004):大黄黄连泻心汤配伍争议的体内外化学成分研究，负责人：周晓虹。