利用UHPLC/Q-TOF MS技术进行知母中7个甾体皂苷的含量测定

利用UHPLC/Q-TOF MS技术进行知母中7个甾体皂苷的含量测定

赵阳, 康利平, 余河水, 庞旭, 王镇方, 熊呈琦, 刘超, 韩立峰, 高秀梅, 马百平^*

1. 北京放射医学研究所, 北京 100850
2. 天津中医药大学, 天津 300193

收稿日期:2013-03-04 修回日期:2013-04-15 出版日期:2013-05-08 发布日期:2013-05-08
通讯作者: 马百平*

Simultaneous determination of steroidal saponins in Anemarrhena asphodeloides Bge. by ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry

Yang Zhao, Liping Kang, Heshui Yu, Xu Pang, Zhenfang Wang, Chengqi Xiong, Chao Liu, Lifeng Han, Xiumei Gao, Baiping Ma^*

1. Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing 100850, China
2. Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China

Received:2013-03-04 Revised:2013-04-15 Online:2013-05-08 Published:2013-05-08
Contact: Baiping Ma*

摘要/Abstract

摘要：

本文利用UHPLC/Q-TOF MS技术建立了知母中7个主要甾体皂苷的含量测定方法, 它们分别为知母皂苷N, 知母皂苷E1, 知母皂苷BII, 知母皂苷B, 知母皂苷I, 知母皂苷A2和知母皂苷AIII。色谱柱为ACQUITY UPLC^® HSS T3 柱(2.1 mm×100 mm, 1.8 μm), 流动相为乙腈-水 (0.1%甲酸) 梯度洗脱, 分析时间为18分钟; LOQ和LOD分别为0.18-0.75 ng/µL和0.05-0.22 ng/µL; 7个甾体皂苷分别在一定范围内呈现良好的线性关系, 相关系数分别为0.9902-0.9979, 且日内及日间精密度均低于5%, 回收率为97.13%-101.98%。该方法快速、准确、重现性好, 适用于知母药材及知母配方颗粒中7个甾体皂苷的含量测定。

关键词: 知母, UHPLC/Q-TOF MS, 甾体皂苷, 含量测定

Abstract:

The objective of the research is to develop a quantitative method by ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) for the analysis of seven major steroidal saponins (timosaponin N, timosaponin E1, timosaponin BII, timosaponin B, anemarrhenasaponin I, anemarrhenasaponin A2, and timosaponin AIII) in Anemarrhena asphodeloides Bge. The complete separation of these seven steroidal saponins was achieved within 18 min with an ACQUITY UPLC HSS T3 column using an acetonitrile-water (contain 0.1% formic acid) gradient system. The limits of quantitation (LOQ), 0.18-0.75 ng/µL for seven steroidal saponins, were determined experimentally. The limits of detection (LOD) were found to be 0.05-0.22 ng/µL for these saponins. The correlation coefficients (r²) for calibration curves varied from 0.9902 to 0.9979. This method showed good repeatability for the quantification of these saponins in rhizomes of A. asphodeloides, with intra-day and inter-day variations of less than 5.0% for seven steroidal saponins. The recoveries of seven steroidal saponins were from 97.13% to 101.98%. The validated method was successfully applied to quantifying seven steroidal saponins in various sources of A. asphodeloides (different collecting places or processing methods) and Zhimu concentrate-granules (ZMCG).

Key words: Anemarrhena asphodeloides Bge., UHPLC/Q-TOF MS, Steroidal saponins, Simultaneous determination

中图分类号:

R284.1

Supporting: Foundation items: Major National Science and Technology Projects (Grant No. 2009ZX09102-106, 2011ZX09102-002-09) and National Natural Science Foundation of China (Grant No. 81274053).
^*Corresponding author. Tel.: 86-10-68210077 ext. 930265

赵阳, 康利平, 余河水, 庞旭, 王镇方, 熊呈琦, 刘超, 韩立峰, 高秀梅, 马百平^*. 利用UHPLC/Q-TOF MS技术进行知母中7个甾体皂苷的含量测定[J]. .

Yang Zhao, Liping Kang, Heshui Yu, Xu Pang, Zhenfang Wang, Chengqi Xiong, Chao Liu, Lifeng Han, Xiumei Gao, Baiping Ma^*. Simultaneous determination of steroidal saponins in Anemarrhena asphodeloides Bge. by ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry [J]. .

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