State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
Nanjing Forestry University, Nanjing 210037, China.
J Pharm Biomed Anal. 2018 May 10;153:152-157. doi: 10.1016/j.jpba.2018.02.019. Epub 2018 Feb 21.
Coptidis Rhizoma (CR), the dried rhizome of three perennial Coptis specices, was widely used as a famous herbal medicine in China. Although the quantification of main alkaloids in CR has been extensively conducted, the existing analytical methods suffer from some flaws that restrict the general applicability in the routine quality assessment. In this work, we constructed an optimized method for quality evaluation and species discrimination of CR by ion-pairing high performance liquid chromatography (IP-HPLC) combined with response surface methodology (RSM). By employing Box-Behnken designs (BBD), 30 sets of experimental runs were performed to build the response surface models, and Derringer's desirability was used to optimize the IP-HPLC separation conditions by simultaneously taking resolutions between two pairs of hardly - separated alkaloids and the retention time of the last eluted analyte as optimization criteria. Meanwhile, a single standard to determine multi-components (SSDMC) method based on the optimized IP-HPLC was set up and fully validated, to simultaneously determine six alkaloids including jatrorrhizine (JAT), columbamine (COL), epiberberine (EPI), coptisine (COP), palmatine (PAL) and berberine (BER), using BER as internal standard. Finally, the quantitative data from 33 batches of CR samples were comparatively analyzed, and the ratios of JAT/COL and EPI/JAT were discovered for species classification. Collectively, the established IP-HPLC method can be adopted for comprehensive quality evaluation and species discrimination of CR due to its general applicability.
黄连（CR）是三种多年生黄连属植物的干燥根茎，在中国被广泛用作著名的草药。虽然已经广泛进行了黄连中主要生物碱的定量分析，但现有的分析方法存在一些缺陷，限制了其在常规质量评估中的普遍适用性。在这项工作中，我们通过离子对高效液相色谱法（IP-HPLC）结合响应面法（RSM）构建了一种优化的黄连质量评价和物种鉴别方法。通过采用 Box-Behnken 设计（BBD），进行了 30 组实验运行，以建立响应面模型，并使用 Derringer 的理想性同时将两对难以分离的生物碱之间的分辨率和最后洗脱分析物的保留时间作为优化标准，对 IP-HPLC 分离条件进行优化。同时，建立了一种基于优化 IP-HPLC 的单标准测定多成分（SSDMC）方法，并对其进行了全面验证，以同时测定 6 种生物碱，包括小檗碱（JAT）、巴马汀（PAL）、黄连碱（COL）、表小檗碱（EPI）、药根碱（JAT）和巴马丁（BER），以 BER 为内标。最后，对 33 批黄连样品的定量数据进行了比较分析，并发现 JAT/COL 和 EPI/JAT 的比值可用于物种分类。总之，由于其普遍适用性，所建立的 IP-HPLC 方法可用于黄连的综合质量评价和物种鉴别。
