Jin Xin, Liu Mingyan, Chen Zaixing, Mao Ruikun, Xiao Qinghuan, Gao Hua, Wei Minjie
School of Pharmacy, China Medical University, Shenyang 110001, China.
School of Pharmacy, China Medical University, Shenyang 110001, China; Division of Pharmacology Laboratory, National Institutes for Food and Drug Control, Beijing 100050, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Oct 1;1002:113-22. doi: 10.1016/j.jchromb.2015.07.055. Epub 2015 Aug 18.
Epigallocatechin-3-gallate (EGCG) is a major bioactive ingredient of green tea that produces beneficial neuroprotective effects. In this paper, to optimize the EGCG enrichment, thirteen macroporous resins with different chemical and physical properties were systemically evaluated. Among the thirteen tested resins, the H-bond resin HPD826 exhibited best adsorption/desorption capabilities and desorption ratio, as well as weakest affinity for caffeine. The absorption of EGCG on the HPD826 resin followed the pseudo-second-order kinetics and Langmuir isotherm model. The separation parameters of EGCG were optimized by dynamic adsorption/desorption experiments with the HPD826 resin column. Under the optimal condition, the content of EGCG in the 30% ethanol eluent increased by 5.8-fold from 7.7% to 44.6%, with the recovery yield of 72.1%. After further purification on a polyamide column, EGCG with 74.8% purity was obtained in the 40-50% ethanol fraction with a recovery rate of 88.4%. In addition, EGCG with 95.1% purity could be easily obtained after one-step crystallization in distilled water. Our study suggests that the combined macroporous resin and polyamide column chromatography is a simple method for large-scale separation and purification of EGCG from natural plants for food and pharmaceutical applications.
表没食子儿茶素没食子酸酯（EGCG）是绿茶的一种主要生物活性成分，具有有益的神经保护作用。本文为优化EGCG的富集，对13种具有不同化学和物理性质的大孔树脂进行了系统评价。在所测试的13种树脂中，氢键树脂HPD826表现出最佳的吸附/解吸能力和解吸率，以及对咖啡因最弱的亲和力。EGCG在HPD826树脂上的吸附遵循准二级动力学和朗缪尔等温线模型。通过HPD826树脂柱的动态吸附/解吸实验优化了EGCG的分离参数。在最佳条件下，30%乙醇洗脱液中EGCG的含量从7.7%提高到44.6%，提高了5.8倍，回收率为72.1%。在聚酰胺柱上进一步纯化后，在40 - 50%乙醇馏分中获得了纯度为74.8%的EGCG，回收率为88.4%。此外，在蒸馏水中一步结晶后可轻松获得纯度为95.1%的EGCG。我们的研究表明，大孔树脂和聚酰胺柱色谱联用是从天然植物中大规模分离纯化EGCG用于食品和医药应用的一种简单方法。
