色谱

色谱 ›› 2017, Vol. 35 ›› Issue (1): 32-46.DOI: 10.3724/SP.J.1123.2016.08025

• 研究论文 • 上一篇    下一篇

超声细胞粉碎萃取结合超高效液相色谱用于六味地黄丸的整体分析(英文)

赵唐娟1,2, 陈娟1, 师彦平1   

  1. 1. 中国科学院兰州化学物理研究所, 中国科学院西北特色植物资源化学重点实验室/甘肃省天然药物重点实验室, 兰州 730000;
    2. 中国科学院大学, 北京 100049
  • 收稿日期:2016-08-23 出版日期:2017-01-08 发布日期:2013-05-06
  • 通讯作者: 赵唐娟,Tel:+86-931-4968121;Fax:+86-931-4968094;E-mail:chenjuan@licp.cas.cn;师彦平,E-mail:shiyp@licp.cas.cn

Holistic analysis of Liuwei Dihuang pills using ultrasonic cell grinder extraction and ultra-performance liquid chromatography

ZHAO Tangjuan1,2, CHEN Juan1, SHI Yanping1   

  1. 1. Key Laboratory of Chemistry of Northwestern Plant Resources of Chinese Academy of Sciences and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China;
    2. University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2016-08-23 Online:2017-01-08 Published:2013-05-06
  • Supported by:

    National Natural Science Foundation of China (Nos. 21375136, 21575150); Scholar Program of West Light Project, Chinese Academy of Sciences.

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摘要：

针对中药复方制剂六味地黄丸的质量控制提出了一种多指标全药材整体分析新策略。从组成六味地黄丸的每一味药材中选取1~2个药效成分，即选取熟地黄中的毛蕊花糖苷、山茱萸中的莫诺苷和马钱苷、牡丹皮中的丹皮酚和芍药苷、山药中的尿囊素、茯苓中的茯苓酸、泽泻中的23-乙酰泽泻醇B作为六味地黄丸的质量控制指标成分，采用快速、简便、高效的超声细胞粉碎提取（UCGE）法对这8个指标成分进行同步提取。用响应曲面分析中的Box-Behnken模式对UCGE提取过程中对提取效率影响较大的料液比、提取时间和提取温度等参数进行了优化。得到最优的提取条件：料液比1：45（g/mL）、提取时间40 min、提取温度20℃。进一步利用超高效液相色谱-二极管阵列检测（UPLC-PAD）技术实现多指标成分在同一根色谱柱上的同步分离和检测，从而建立六味地黄丸的多指标全药材整体色谱。采用HSS T3色谱柱（50 mm×2.1 mm，1.8 μm），以乙腈-水为流动相进行梯度洗脱，基于指标成分的紫外光谱特征选择5个检测波长同时检测，并进行了线性关系、检出限、定量限、精密度、稳定性、重复性、准确性等方法学验证。该研究为六味地黄丸质量标准的建立提供了依据，同时也为中药复方制剂提供了一种质量控制新模式和新理念。

关键词: 超声细胞粉碎提取, 六味地黄丸, 响应曲面法, 质量控制

Abstract:

An efficient holistic analysis strategy was developed and validated for quality control of the traditional Chinese medicine (TCM) preparation, Liuwei Dihuang pills (LWDHPs). In this strategy, eight bioactive components, i. e. allantoin, morroniside, loganin, peoniflorin, acteoside, paeonol, alisol B 23-acetate and pachymic acid from the six medicinal herbs composing LWDHPs were selected as the evaluation markers, and then they were simultaneously extracted by an ultrasonic cell grinder extraction (UCGE) method and separated in a single run by an ultra-performance liquid chromatographic method coupled with photodiode array detection (UPLC-PDA). Response surface methodology (RSM), a multivariate experimental design technique, was used to optimize the UCGE parameters based on Box-Behnken design (BBD) model. The optimum extraction conditions were obtained at a ratio of solvent to solid 45 mL/g, extraction time of 40 min and extraction temperature of 20℃. Under such conditions, the higher yields of the eight analytes were acquired compared with the traditional extraction methods, indicating that UCGE was a fast, easy and efficient method for extracting bioactive constituents from LWDHPs. Chromatographic separation was achieved on an HSS T3 column (50 mm×2.1 mm, 1.8 μm) using a linear gradient elution of acetonitrile and water. Because of the different UV characteristics of these components, five detection wavelengths were used for quantitative analysis. The proposed method was validated through linearity, limits of detection, limits of quantification, precision, stability, repeatability, and accuracy. The validated method was applied to analyze LWDHPs, which provided a reference for the quality evaluation of LWDHPs.

Key words: Liuwei Dihuang pills (LWDHPs), quality control, response surface methodology (RSM), ultrasonic cell grinder extraction (UCGE)

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