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HPLC-QTOF-MS/MS结合NMR分析甘草酸单铵盐原料及其高温强降解杂质*

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  • 1.江苏省中药资源产业化过程协同创新中心, 南京 210023;
    2.南京中医药大学药学院, 南京 210023;
    3.新疆医科大学药学院, 乌鲁木齐 830011
第一作者 Tel:(025)86798186; E-mail:zyleeyang@163.com
**Tel:(025)86798186; E-mail:zyunfeng@njucm.edu.cn

收稿日期: 2020-11-20

  网络出版日期: 2024-06-21

基金资助

*国家自然科学基金项目(81973482);新疆维吾尔自治区重点研发专项(2020B03007-2);国家大学生创新创业训练计划(202010315033)

Identification and characterization of the related substances of monoammonium glycyrrhizinate and its high temperature degradation by HPLC-QTOF MS/MS and NMR*

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  • 1. Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing 210023, China;
    2. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China;
    3. College of Pharmacy, Xinjiang Medical University, Urumqi 830011, China

Received date: 2020-11-20

  Online published: 2024-06-21

摘要

目的:分析鉴定甘草酸单铵盐原料及其高温强降解有关杂质成分。方法:采用Shim-pack GIST C18色谱柱(150 mm×4.6 mm,5 μm),以乙腈-1%乙酸水溶液(34.4∶65.6)为流动相,对甘草酸单铵盐及高温强降解原料中的有关物质进行分离,采用四级杆飞行时间质谱进行检测,结合其分子离子峰、碎片离子及裂解规律进行分析,鉴定其化学结构;进一步采用C18中压柱(800 g,25~50 μm,6.5 cm×50 cm),以甲醇-水-乙酸(40∶60∶1、50∶50∶1、60∶40∶1)洗脱,分离制备有关物质,并用核磁共振波谱(1H-NMR、13C-NMR、HSQC、HMBC)对其中2个未知杂质成分进行结构确证。结果:通过对照品对照及HPLC-QTOF-MS/MS分析,鉴定了甘草酸单铵盐及其高温强降解原料中13个三萜皂苷类杂质成分,其中2个未知杂质成分经鉴定分析,结构确证为6′-甲基甘草酸及6″-甲基甘草酸。结论:HPLC-QTOF-MS/MS法可以快速分析鉴定甘草酸单铵盐中的未知杂质,可为其原料及制剂杂质限度的制定奠定基础。

本文引用格式

李洋, 孙捷, 段伟萍, 高晓黎, 赵金兰, 谢湘云, 彭国平, 郑云枫 . HPLC-QTOF-MS/MS结合NMR分析甘草酸单铵盐原料及其高温强降解杂质*[J]. 药物分析杂志, 2021 , 41(10) : 1748 -1756 . DOI: 10.16155/j.0254-1793.2021.10.12

Abstract

Objective: To analyze and identify the related substances of monoammonium glycyrrhizinate and its high temperature degradation. Methods: The separation of monoammonium glycyrrhizinate and its high temperature degradation were carried out on Shim-pack GIST C18(150 mm×4.6 mm,5 μm) column, using 0.1% formic acid water and acetonitrile (34.4∶65.6) as the mobile phase. The quadrupole time-of-flight mass spectrometry was used for detection and identification, combing the molecular ion peaks, fragment ions and fragmentation pattern. And the impurities were then separated and purified by a C18 medium pressure colum (800 g, 25-50 μm, 6.5 cm×50 cm, washing with methanol, water and acetic acid (40∶60∶1, 50∶50∶1, 60∶40∶1). The structures of two unknown impurities were confirmed by nuclear magnetic resonance(1H-NM R, 13C-NMR, HSQC, HMBC). Results: Thirteen related substances of triterpenoid saponins were identified, and two unknown impurities of them were clarified as 6′-methyl-glycyrrhizin and 6″-methyl-glycyrrhizin. Conclusion: The established method using HPLC-QTOF MS/MS can quickly detect and analyze the impurities in monoammonium glycyrrhizinate, which can be used for the quality control of the bulk drug and preparations of monoammmonium glycyrrhizinate.

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