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采用L9(34)正交优化和层析分离、纯化EGCG,研究不同层析介质、洗脱剂和洗脱剂的酸度、浓度及流速对茶多酚中EGCG分离、纯化效果的影响。结果表明:影响EGCG分离、纯化因素的主次顺序为:层析介质﹥洗脱剂﹥洗脱剂pH值﹥洗脱剂浓度;方差分析显示,4种因素对EGCG的分离、纯化影响均不显著。应用单因素试验检验正交实验,结果表明,采用聚酰胺作层析介质,洗脱剂为丙酮,洗脱剂pH3.5,洗脱剂流速1.0mL/min时效果最佳。另外,在聚酰胺层析分离试验中,EGCG流出集中在85～150min之间,100min时EGCG洗脱达最大值,此时可获高纯度的EGCG单体。
EGCG was purified by orthogonal optimization of L9 (34) and chromatographic separation. The effects of different chromatographic media, eluent and eluant acidity, concentration and flow rate on the separation and purification of EGCG in tea polyphenols were studied. The results showed that the main order of the factors influencing the separation and purification of EGCG was as follows: chromatographic medium, eluant, eluent, pH, and eluent concentration. Analysis of variance showed that the effects of four factors on the separation and purification of EGCG were not Significant. The single factor test was used to test the orthogonal experiment. The results showed that polyamide was the best chromatographic medium with acetone as the eluent, pH3.5 as the eluent, and the flow rate of the eluent as 1.0 mL / min. In addition, in the polyamide chromatographic separation test, EGCG efflux concentrated between 85 ~ 150min, EGCG 100min when the maximum elution, high-purity EGCG monomer available at this time.