逆流色谱法分离纯化蒲公英中多酚类化合物

doi:10.3976/j.issn.1002-4026.2022.06.001

山东科学 ›› 2022, Vol. 35 ›› Issue (6): 1-7.doi: 10.3976/j.issn.1002-4026.2022.06.001

• 中药与天然活性产物 • 下一篇

逆流色谱法分离纯化蒲公英中多酚类化合物

高洁¹^,²^,³(), 宋祥云¹, 马天宇¹, 刘峰¹, ROMAN Kachan³^,^*(), 王晓¹^,^*()

1.齐鲁工业大学(山东省科学院) 山东省分析测试中心,山东济南 250014
2.齐鲁工业大学(山东科学院) 基辅学院,山东济南 250353
3.基辅国立工艺设计大学,乌克兰基辅 01011

收稿日期:2021-11-19 出版日期:2022-12-20 发布日期:2022-12-02
通信作者: ROMAN Kachan,王晓 E-mail:alice8600@163.com;kachanr@gmail.com;wangx@sdas.org
作者简介:高洁(1995—),女,硕士研究生,研究方向为天然产物研究与开发。E-mail:alice8600@163.com
基金资助:
山东省重点研发计划(重大科技创新工程)(2021CXGC010508);山东省自然科学基金面上项目(ZR2020MH397)

Separation and purification of polyphenols from Taraxaci Herba by counter-current chromatography

GAO Jie¹^,²^,³(), SONG Xiang-yun¹, MA Tian-yu¹, LIU Feng¹, ROMAN Kachan³^,^*(), WANG Xiao¹^,^*()

1. Shandong Analysis and Test Center, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250014, China
2. Kiev College, Qilu University of Technology (Shandong Academy of Sciences),Jinan 250353, China
3. Kyiv National University of Technologies and Design, Kiev 01011, Ukraine

Received:2021-11-19 Online:2022-12-20 Published:2022-12-02
Contact: ROMAN Kachan, WANG Xiao E-mail:alice8600@163.com;kachanr@gmail.com;wangx@sdas.org

摘要/Abstract

摘要：

采用pH区带逆流色谱(pH-zone-refining counter-current chromatography,pH-ZRCCC)与高速逆流色谱(high-speed counter-current chromatography,HSCCC)相结合的方式,从蒲公英中分离得到6个多酚类化合物。在pH-ZRCCC分离中,选择溶剂体系为乙酸乙酯-乙腈-水(体积比4:1:5),上相加入三氟乙酸(10 mmol/L)作为固定相,下相加入氨水(10 mmol/L)作为流动相。从1.6 g蒲公英乙酸乙酯粗提物中分离得到咖啡酸(60.2 mg,纯度98.1%)和对羟基肉桂酸(6.3 mg,纯度98.8%)及混合物A(590 mg)。之后进一步利用HSCCC,选择溶剂体系为石油醚-乙酸乙酯-甲醇-水(体积比1:4:1:4)。从400 mg混合物A中分离得到1-O-咖啡酰基甘油(7.7 mg,纯度82.2%)、3,4-二羟基苯乙酸(5.4 mg,纯度24.8%)、原儿茶酸(6.2 mg,纯度95.3%)以及对羟基苯乙酸(3.4 mg,纯度89.0%)。该方法制备量大,重现性好,适合蒲公英多酚类化合物的分离纯化。

关键词: 蒲公英, 多酚, 高速逆流色谱, pH区带逆流色谱, 分离纯化

Abstract:

Six polyphenolic compounds were separated and extracted from Taraxaci Herba through a combination of pH-zone-refining counter-current chromatography (pH-ZRCCC) and high-speed counter-current chromatography (HSCCC). In pH-ZRCCC,ethyl acetate-acetonitrile-water (4:1:5, V/V) was selected as the solvent system, trifluoroacetic acid (10 mmol/L) was added to the upper phase as the stationary phase, and ammonia (10 mmol/L) was added to the lower phase as the mobile phase. Caffeic acid (60.2 mg with a purity of 98.1%), p-hydroxycinnamic acid (6.3 mg with a purity of 98.8%), and mixture A (590 mg) were separated from 1.6 g of crude ethyl acetate extract of Herba Taraxaci. After further using HSCCC, petroleumether-ethyl acetate-methanol-water (1:4:1:4, V/V) was selected as the solvent system.Consequently, 1-O-caffeoylglycerol (7.7 mg with a purity of 82.2%), 3,4-dihydroxyphenylacetic acid (5.4 mgwith a purity of 24.8%), protocatechuic acid (6.2mg with a purity of 95.3%) and p-hydroxyphenylacetic acid (3.4 mgwith a purity of 89.0%) were obtained from 400 mg of mixture A.The method has a large preparation volume and good reproducibility, and is suitable for the separation and purification of Taraxaci Herba polyphenol compounds.

Key words: Taraxaci Herba, polyphenols, high-speed counter current chromatography, pH-zone-refining counter-current chromatography, separation and purification

中图分类号:

R284.2

高洁, 宋祥云, 马天宇, 刘峰, ROMAN Kachan, 王晓. 逆流色谱法分离纯化蒲公英中多酚类化合物[J]. 山东科学, 2022, 35(6): 1-7.

GAO Jie, SONG Xiang-yun, MA Tian-yu, LIU Feng, ROMAN Kachan, WANG Xiao. Separation and purification of polyphenols from Taraxaci Herba by counter-current chromatography[J]. Shandong Science, 2022, 35(6): 1-7.

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时间/min	流量/mL	甲醇体积分数(A)/%	0.1%甲酸水体积分数(B)/%
0.01	1.00	20.0	80.0
10.00	1.00	24.0	76.0
20.00	1.00	24.0	76.0
21.00	1.00	34.0	66.0
30.00	1.00	40.0	60.0
35.00	1.00	100.0	0

分配系数	目标化合物
分配系数	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Ⅴ	Ⅵ
K_acid	29.92	6.43	9.29	4.61	0.20	1.94
K_base	0.00	0.75	0.00	0.05	0.02	0.00

逆流色谱法分离纯化蒲公英中多酚类化合物

Separation and purification of polyphenols from Taraxaci Herba by counter-current chromatography

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