逆流色谱法分离纯化蒲公英中多酚类化合物

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。

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

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.