Song Zhibo, Chen Tao, Wang Shuo, Shen Cheng, Ma Yumei, Li Aijing, Chen Zhi, Li Yulin
Northwest Institute of Plateau Biology, Chinese Academy of Science, Xining, P. R. China.
Savaid Medical School, University of the Chinese Academy of Sciences, Beijing, P. R. China.
J Sep Sci. 2023 Jan;46(2):e2200748. doi: 10.1002/jssc.202200748. Epub 2022 Nov 14.
The separation of polar compounds is challenging work due to poor retention and insufficient selectivity. In the present study, an efficient strategy for large-scale preparation of five polar polyphenols including three isomers from Phyllanthus emblica Linn has been established by preparative high-speed counter-current chromatography. Macroporous resin column chromatography was used for the enrichment of the polar polyphenols. However, sugar and other ultra-polar impurities were co-washed out with the targets. Liquid-liquid extraction with ethyl acetate/water (1/1, v/v) solvent system was developed to remove the ultra-polar impurities with a clearance rate of 95%. Finally, the targets were introduced to preparative high-speed counter-current chromatography for separation using ethyl acetate/n-butanol/acetic acid/water (2/7/1/10, v/v/v/v) solvent system. As a result, 191 mg of Mucic acid 1,4-lactone 5-O-gallate, 370 mg of β-Glucogallin, 301 mg of Gallic acid, 195 mg of Mucic acid 1,4-lactone 3-O-gallate and 176 mg of Mucic acid 1,4-lactone 2-O-gallate with purity higher than 98% were obtained from 1.5 g of sample. Mucic acid 1,4-lactone 3-O-gallate, Mucic acid 1,4-lactone 3-O-gallate, and Mucic acid 1,4-lactone 2-O-gallate are isomers. The results showed that high-speed counter-current chromatography could be well developed for the separation of polar compounds from natural products.
由于保留性差和选择性不足，极性化合物的分离是一项具有挑战性的工作。在本研究中，通过制备型高速逆流色谱法建立了一种从余甘子中大规模制备包括三种异构体在内的五种极性多酚的有效策略。大孔树脂柱色谱法用于富集极性多酚。然而，糖和其他超极性杂质与目标物一起被洗脱出来。开发了用乙酸乙酯/水（1/1，v/v）溶剂体系进行液液萃取以去除超极性杂质，清除率为95%。最后，将目标物引入制备型高速逆流色谱法中，使用乙酸乙酯/正丁醇/乙酸/水（2/7/1/10，v/v/v/v）溶剂体系进行分离。结果，从1.5 g样品中获得了191 mg纯度高于98%的粘酸1,4-内酯5-O-没食子酸酯、370 mgβ-葡萄糖没食子酸酯、301 mg没食子酸、195 mg粘酸1,4-内酯3-O-没食子酸酯和176 mg粘酸1,4-内酯2-O-没食子酸酯。粘酸1,4-内酯3-O-没食子酸酯、粘酸1,4-内酯3-O-没食子酸酯和粘酸1,4-内酯2-O-没食子酸酯是异构体。结果表明，高速逆流色谱法可很好地用于从天然产物中分离极性化合物。
