高速逆流色谱结合半制备液相色谱制备甘青青兰中3种活性成分

doi:10.7525/j.issn.1673-5102.2022.02.010

植物研究 ›› 2022, Vol. 42 ›› Issue (2): 252-258.doi: 10.7525/j.issn.1673-5102.2022.02.010

高速逆流色谱结合半制备液相色谱制备甘青青兰中3种活性成分

周加本¹, 热增才旦², 热毛先¹, 利毛才让¹^,³()

^1.青海师范大学生命科学学院，西宁 810008
^2.青海民族大学药学院，西宁 810007
^3.青海师范大学民族师范学院，西宁 810008

收稿日期:2021-03-18 出版日期:2022-03-20 发布日期:2022-02-22
通讯作者: 利毛才让 E-mail:1224863256@qq.com
作者简介:周加本（1996—），男，硕士研究生，主要从事药用植物化学方面的研究。
基金资助:
国家自然科学基金(31660086);青海省青藏高原药用动植物资源重点实验室(2017-ZJ-Y13)

Separation and Purification of Three Active Components from Dracocephalum tanguticum Maxim. by High-Speed Counter-current Chromatography Combined with Semi-Preparative High Performance Liquid Chromatography

Jiaben Zhou¹, Caidan Rezeng², Maoxian Re¹, Cairang Limao¹^,³()

^1.College of Life Sciences，Qinghai Normal University，Xining 810008
^2.College of Pharmacy，Qinghai University for Nationalities，Xining 810007
^3.College of Nationalities，Qinghai Normal University，Xining 810008

Received:2021-03-18 Online:2022-03-20 Published:2022-02-22
Contact: Cairang Limao E-mail:1224863256@qq.com
About author:Zhou Jiaben（1996—），male，postgraduate，research direction for Medicinal Phytochemistry．
Supported by:
National Natural Science Foundation of China(31660086);Key Laboratory of Medicinal Animal and Plant Resources in Qinghai-Tibet Plateau of Qinghai Province(2017-ZJ-Y13)

摘要/Abstract

摘要：

采用高速逆流色谱（HSCCC）结合液相色谱法制备甘青青兰（Dracocephalum tanguticum Maxim.）中绿原酸、胡麻甙-6″-乙酯、迷迭香酸，建立快速分离制备甘青青兰中活性成分的方法。采用半制备型高效液相色谱（SP-HPLC）富集甘青青兰乙酸乙酯萃取物中绿原酸、胡麻甙-6″-乙酯、迷迭香酸，再用制备液相（Pre-HPLC）和HSCCC对富集物进行分离纯化，获得54 mg化合物Ⅰ、130 mg化合物Ⅱ和200 mg化合物Ⅲ，纯度分别为96.9%、97.9%和95.1%，经核磁共振碳谱（¹³CNMR）和氢谱（¹HNMR）分别鉴定为绿原酸、胡麻甙-6″-乙酯和迷迭香酸。本实验方法适用于甘青青兰乙酸乙酯萃取物中绿原酸、胡麻甙-6″-乙酯和迷迭香酸的分离制备，并避免了传统分离方法操作繁多、试剂消耗量大、不可回收等弊端，为分离甘青青兰活性成分、制备对照品等研究提供了参考依据。

关键词: 高速逆流色谱, 半制备型液相色谱, 甘青青兰, 绿原酸, 胡麻甙-6″-乙酯, 迷迭香酸

Abstract:

An efficient method was established for the separation and preparation of chlorogenic acid， pedaliin-6″-acetate and rosmarinic acid from Dracocephalum tanguticum Maxim.. The chlorogenic acid， pedaliin-6″-acetate and rosmarinic acid was enriched by semi-preparative high performance liquid chromatography（SP-HPLC） from ethyl acetate extract， and then separated and purification by high-speed counter-current chromatography（HSCCC） and preparative high performance liquid chromatography（Pre-HPLC）. A total of 54 mg of chlorogenic acid， 150 mg of pedaliin-6″- acetate and 200 mg of rosmarnic acid were obtained from ethyl acetate extract with purities of 96.9%， 97.9% and 95.1% by HPLC analysis， respectively. Their structures were identified by nuclear magnetic resonance spectrum. The result show that this method is suitable for the isolation and purification of chlorogenic acid， pedaliin-6″-acetate and rosmarnic acid from D. tanguticum. It also avoids the disadvantages of traditional separation methods such as many operations， large reagent consumption and non-recyclability， provides a reference basis for the study of separating of active components and preparation reference substance from D.tanguticum.

Key words: high-speed counter-current chromatography, preparative high-performance liquid chromatography, Dracocephalumtanguticum Maxim., chlorogenic acid, Pedaliin-6″-acetate, rosmarnic acid

中图分类号:

S132

周加本, 热增才旦, 热毛先, 利毛才让. 高速逆流色谱结合半制备液相色谱制备甘青青兰中3种活性成分[J]. 植物研究, 2022, 42(2): 252-258.

Jiaben Zhou, Caidan Rezeng, Maoxian Re, Cairang Limao. Separation and Purification of Three Active Components from Dracocephalum tanguticum Maxim. by High-Speed Counter-current Chromatography Combined with Semi-Preparative High Performance Liquid Chromatography[J]. Bulletin of Botanical Research, 2022, 42(2): 252-258.

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溶剂体系 Solvent system	体积配比 Volume ratio	分配系数K		分离度 R
溶剂体系 Solvent system	体积配比 Volume ratio	Ⅱ	Ⅲ	分离度 R
乙酸乙酯—正丁醇—水 Ethyl acetate—n-butanol—water	4∶1∶5	4.92	3.20	0.65
正己烷—乙酸乙酯—甲醇—水 N-hexane—ethyl acetate—methanol—water	1∶3∶1∶3	0.10	0.57	5.61
	1∶5∶1∶5	0.22	0.96	4.30
	1∶7∶1∶7	0.38	1.43	3.82
	1∶9∶1∶9	0.40	1.48	3.74
	2∶3∶2∶3	0.02	0.06	2.75
	2∶5∶2∶5	0.09	0.42	4.43
	1∶10∶1∶10	0.60	2.32	3.86
	1∶12∶1∶12	0.69	2.10	4.67
	1∶14∶1∶1∶14	0.81	2.97	3.68
正己烷—乙酸乙酯—正丁醇—水 N-hexane—ethyl acetate—n-butanol—water	1∶9∶1∶1∶9	1.72	5.19	3.00
石油醚—乙酸乙酯—甲醇—水 Petroleum ether—ethyl acetate—methanol—water	1∶12∶1∶12	1.27	2.70	2.13

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高速逆流色谱结合半制备液相色谱制备甘青青兰中3种活性成分

Separation and Purification of Three Active Components from Dracocephalum tanguticum Maxim. by High-Speed Counter-current Chromatography Combined with Semi-Preparative High Performance Liquid Chromatography

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