正交设计试验优化高速逆流色谱分离制备玛咖中芥子油苷的条件

doi:10.3724/SP.J.1123.2016.03024

色谱 ›› 2016, Vol. 34 ›› Issue (8): 788-794.DOI: 10.3724/SP.J.1123.2016.03024

正交设计试验优化高速逆流色谱分离制备玛咖中芥子油苷的条件

王亚钦¹, 吴迪², 赵学志¹, 朱尘琪², 郭爽², 高杨亚雅², 邱百灵², 宋宇², 韩南银², 何洪巨¹

1. 北京市农林科学院蔬菜研究中心, 北京 100097;
2. 北京大学药学院, 北京 100191

收稿日期:2016-03-16 出版日期:2016-08-08 发布日期:2016-07-26
通讯作者: 何洪巨, 韩南银
基金资助:
农业部公益性行业专项项目（20130309）；北京市农林科学院科技创新能力建设专项（KJCX20140111）.

Optimization of conditions for the isolation and preparation of glucosinolates from Maca by high-speed counter-current chromatography with orthogonal experiments

WANG Yaqin¹, WU Di², ZHAO Xuezhi¹, ZHU Chenqi², GUO Shuang², GAOYANG Yaya², QIU Bailing², SONG Yu², HAN Nanyin², HE Hongju¹

1. Vegetable Research Center, Beijing Municipal Academy of Agriculture and Forestry, Beijing 100097, China;
2. School of Pharmaceutical Sciences, Peking University, Beijing 100191, China

Received:2016-03-16 Online:2016-08-08 Published:2016-07-26
Supported by:
Research Projects of Agricultural Public Welfare Industry in China (No. 20130309); Innovation and Capacity-building Projects by Beijing Academy of Agriculture and Forestry Sciences (No. KJCX20140111).

摘要/Abstract

摘要：

基于高速逆流色谱（HSCCC）技术从玛咖中分离制备出两种芥子油苷，苄基芥子油苷（glucotropaeolin, GTL）和甲氧基苄基芥子油苷（glucolimnanthin, GLI）。使用正交设计试验对分离条件进行优化，采用高分辨质谱对制备的组分进行鉴定，采用高效液相色谱法（HPLC）对组分进行定量分析。确定了两个组分GTL与GLI的HSCCC最佳分离条件：溶剂系统为正丁醇-乙腈-200 g/L硫酸铵溶液（1:0.5:2.4, v/v/v），上相为固定相，下相为流动相，流动相流速2 mL/min，主机转速900 r/min，从玛咖根粗提物中一次性分离得到157.72 mg/kg纯度为97.9%的苄基芥子油苷和31.93 mg/kg的甲氧基苄基芥子油苷，固定相保留率达57.6%。该方法成本低，简便易行，样品损失量小，可大量循环进样制备。

关键词: 高速逆流色谱, 芥子油苷, 玛咖, 正交设计试验

Abstract:

Glucosinolate is an important bioactive substance in Maca. In our study, two glucosinolates named glucotropaelin (GTL) and glucolimnanthin (GLI) were successfully separated from Maca with semi-preparative high-speed counter-current chromatography (HSCCC). For a best separation and better stationary phase retention, orthogonal experiments were carried out with the flow-rate of the mobile phase, ammonium sulfate concentration of biphasic solvent system, injection volume of crude extracts and the centrifugal speed of the apparatus as the four main factors. The separation was performed with the biphasic solvent system composed of normal butanol-acetonitrile-200 g/L ammonium sulfate solution (1:0.5:2.4, v/v/v) and the upper phase was used as the stationary phase and the lower phase as the mobile phase. The detection wavelength was set at 229 nm. The obtained optimum preparation conditions were that the mobile phase flow-rate was 2 mL/min, 500.00 mg crude extract was dissolved in 15 mL mobile phase, the ammonium sulfate concentration was 200 g/L and the centrifugal speed of the apparatus was 900 r/min. Under these conditions, a high efficiency separation of HSCCC was achieved, and 157.72 mg/kg of GTL and 31.93 mg/kg of GLI were obtained from the crude sample with a high stationary phase retention of 57.6%. The high performance liquid chromatography (HPLC) analysis showed that the purity of GTL was 97.9%. The identification and quantification of glucosinolates were completed by HPLC and mass spectrometry (MS). The established separation method is simple, less adsorption loss, lower cost and suitable for large scale separation of glucosinolates from Maca.

Key words: Lepidium meyenii Walp, glucosinolates, high-speed counter-current chromatography (HSCCC), orthogonal experiments

中图分类号:

O658

王亚钦, 吴迪, 赵学志, 朱尘琪, 郭爽, 高杨亚雅, 邱百灵, 宋宇, 韩南银, 何洪巨. 正交设计试验优化高速逆流色谱分离制备玛咖中芥子油苷的条件[J]. 色谱, 2016, 34(8): 788-794.

WANG Yaqin, WU Di, ZHAO Xuezhi, ZHU Chenqi, GUO Shuang, GAOYANG Yaya, QIU Bailing, SONG Yu, HAN Nanyin, HE Hongju. Optimization of conditions for the isolation and preparation of glucosinolates from Maca by high-speed counter-current chromatography with orthogonal experiments[J]. Chinese Journal of Chromatography, 2016, 34(8): 788-794.

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摘要

正交设计试验优化高速逆流色谱分离制备玛咖中芥子油苷的条件

Optimization of conditions for the isolation and preparation of glucosinolates from Maca by high-speed counter-current chromatography with orthogonal experiments

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