色谱 ›› 2021, Vol. 39 ›› Issue (9): 989-997.DOI: 10.3724/SP.J.1123.2021.06019

• 研究论文 • 上一篇    下一篇

基于高效液相色谱定量指纹图谱和液相色谱-质谱联用定量的酸枣仁提取物质量考察

郭秀洁1,2, 李昊虬3,4, 冯昊天3,4, 戚华文2, 张露2, 徐伟2, 伍言娟2, 王超然1,2,*(), 梁鑫淼1,2   

  1. 1.中国科学院大连化学物理研究所, 辽宁 大连 116023
    2.中科院大化所中国医药城生物医药创新研究院, 江苏 泰州 225300
    3.内蒙古乳业技术研究院有限责任公司, 内蒙古 呼和浩特 010110
    4.内蒙古伊利实业集团股份有限公司, 内蒙古 呼和浩特 010110
  • 收稿日期:2021-06-17 出版日期:2021-09-08 发布日期:2021-09-06
  • 通讯作者: 王超然
  • 作者简介:* Tel:(0523)86205601,E-mail: wcrpj_505@dicp.ac.cn.
  • 基金资助:
    江苏省双创团队项目

Quality analysis of Ziziphi Spinosae Semen extracts based on high performance liquid chromatography quantitative fingerprint and ultra-high performance liquid chromatography-tandem mass spectrometry quantification

GUO Xiujie1,2, LI Haoqiu3,4, FENG Haotian3,4, QI Huawen2, ZHANG Lu2, XU Wei2, WU Yanjuan2, WANG Chaoran1,2,*(), LIANG Xinmiao1,2   

  1. 1. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
    2. DICP-CMC Innovation Institute of Medicine, Taizhou 225300, China
    3. Inner Mongolia Dairy Technology Research Institute Co. Ltd., Huhehaote 010110, China
    4. Inner Mongolia Yili Industrial Group Co. Ltd., Huhehaote 010110, China
  • Received:2021-06-17 Online:2021-09-08 Published:2021-09-06
  • Contact: WANG Chaoran
  • Supported by:
    High-Level Innovative Talent Team of Jiangsu Province

摘要:

酸枣仁具有显著的改善睡眠和抗焦虑等作用,其提取物在助眠类功能食品开发中应用前景广阔。但目前市场上酸枣仁提取物质量参差不齐,缺乏统一标准,企业在使用时面临较大的质量风险,因此亟须建立一种准确、全面的内控质量评价方法。针对酸枣仁提取物中黄酮和皂苷两类主要活性成分紫外响应差异巨大且水提物中皂苷成分含量低的问题,该研究建立了酸枣仁水提物HPLC定量指纹图谱方法,共标定了8个共有峰。通过对照品指认、文献比对以及高效液相色谱-四极杆飞行时间质谱数据解析,8个共有峰均为黄酮类化合物,该方法可同时实现7种黄酮成分的半定量对比分析和斯皮诺素的含量测定;采用超高效液相色谱-三重四极杆质谱,在正离子模式下,以多反应监测扫描方式可实现酸枣仁皂苷A和B的含量测定;最终以雷达图展现上述10种成分的半定量和定量数据。应用上述方法,该研究对比分析了实验室自制的3批酸枣仁水提物和15家供应商的15批提取物样品。结果显示,实验室自制的3批酸枣仁水提物虽然原料来自不同饮片企业,但总体差异性不大,而不同厂家提供的酸枣仁提取物样品成分含量差异巨大,提示不同厂家存在辅料稀释、理枣仁掺假和醇提或纯化富集等情况。该法为企业制定内控质量标准和筛选合格供应商提供了依据。

关键词: 高效液相色谱, 质谱, 定量指纹图谱, 酸枣仁提取物, 雷达图

Abstract:

Ziziphi Spinosae Semen is the dried seeds of Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou, and its extract has broad application prospects in the development of sleep-aid functional foods. However, the quality parameters of Ziziphi Spinosae Semen extracts currently available in the market are not uniform and there is a lack of unified standards. Therefore, it is important to establish an accurate and comprehensive method for quality evaluation. In view of the problems that the UV responses of flavonoids and saponins in the Ziziphi Spinosae Semen extracts vary dramatically and the saponin content in Ziziphi Spinosae Semen water extract is very low, high performance liquid chromatography (HPLC) was used to establish the fingerprint and quantify spinosin. The separation was carried out on a Waters XSelect HSS C18 column (250 mm×4.6 mm, 5 μm), and the mobile phase was acetonitrile-0.1% (v/v) phosphoric acid aqueous solution for gradient elution. The eight common peaks in the fingerprint of the Ziziphi Spinosae Semen extracts, identified by HPLC-quadrupole time-of-flight mass spectrometry, were attributed to flavonoids by reference substance identification, literature comparison, and high-resolution mass spectrometry data analysis. Semi-quantitative analysis of seven flavonoids and quantitative analysis of spinosin were conducted using the established HPLC quantitative fingerprint. The contents of jujuboside A and jujuboside B were determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) by gradient elution using a mobile phase of acetonitrile-0.1%(v/v) formic acid aqueous solution. The target compounds were analyzed in multiple reaction monitoring mode with positive electrospray ionization.
The semi-quantitative and quantitative data of the above-mentioned 10 components are displayed in the form of radar. Using the above methods, three batches of Ziziphi Spinosae Semen water extracts prepared in the laboratory and 15 batches of extract samples obtained from 15 suppliers were analyzed and compared. The results showed that although the raw materials of three batches of Ziziphi Spinosae Semen water extracts prepared in the laboratory were from different enterprises, the overall difference was not significant. However, the component contents of the samples provided by different manufacturers were greatly different, suggesting that there are some problems associated with the different manufacturers, such as dilution of excipients, adulteration of Ziziphi Mauritianae Semen, alcohol extraction, purification, and enrichment. For example, the representative composition contents in the Ziziphi Spinosae Semen extracts obtained from manufacturers B, C, E, F, G, H, I, and O were low, which were approximately 1/10 of corresponding contents in the normal water extracts prepared in the laboratory. It is speculated that to reduce the unit price of the product, the manufacturer used fewer raw materials or a large number of auxiliary materials to dilute the Ziziphi Spinosae Semen extracts. The contents of some flavonoids in the Ziziphi Spinosae Semen extract from manufacturer N were slightly higher than that in the self-preparation Ziziphi Spinosae Semen water extract, but it did not contain jujuboside A; thus, it was speculated that the Ziziphi Mauritianae Semen might be used for extraction. The contents of 10 components in the Ziziphi Spinosae Semen extract obtained from manufacturer D were all higher than the corresponding ones in the self-preparation Ziziphi Spinosae Semen water extract. Combined with the quality label of total saponin content > 20% and poor water solubility, it was speculated that the product might be prepared by alcohol extraction or purified and enriched by using resin. These results provided the basis for the enterprise to establish internal control quality standards for Ziziphi Spinosae Semen extracts and to select qualified suppliers.

Key words: high performance liquid chromatography (HPLC), mass spectrometry (MS), quantitative fingerprint, Ziziphi Spinosae Semen extracts, radar-gram

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