Key Laboratory of Particle and Radiation Imaging, Ministry of Education, Department of Engineering Physics, Tsinghua University, Beijing 100084, P.R. China.
China Pharmaceutical Preparation Section, Huazhong University of Science and Technology Union Jiangbei Hospital/Wuhan Caidian People's Hospital, Wuhan 430100, P.R. China.
J Chromatogr Sci. 2023 Jul 9;61(6):539-545. doi: 10.1093/chromsci/bmac020.
In this study, secondary metabolites of Eurotium cristatum were isolated and purified by high-speed counter-current chromatography (HSCCC), and their hypoglycemic activities were studied. The general-useful estimate of solvent systems (GUESS) for counter-current chromatography was employed to select the appropriate solvent systems of n-hexane-ethyl acetate-methanol-water (HEMW, 4:6:5:5, v/v/v/v) for HSCCC practice, and three compounds were separated from the crude ethyl acetate extract of E. cristatum in one single step; 6.1 mg of Compounds 1, 5.6 mg of Compound 2 and 3.8 mg of Compound 3 were obtained from 100 mg of crude extract with a stationary phase retention of 75%. The compounds were then identified as emodin methyl ether, chrysophanol and emodin, respectively. The activity of the target compounds in the secondary metabolites of E. cristatum was verified by testing their inhibition on α-glucosidase activity and molecular docking simulation. The results showed that emodin, chrysophanol and emodin methyl ether had significant inhibitory effects on the α-glucosidase activity. This work confirmed the effectiveness of HSCCC in the separation of compounds in complex extracts and provided reference for further research and application of E. cristatum.
在这项研究中，采用高速逆流色谱（HSCCC）对青霉次级代谢产物进行了分离和纯化，并研究了其降血糖活性。采用通用溶剂估计（GUESS）法选择合适的溶剂系统（n-己烷-乙酸乙酯-甲醇-水，4:6:5:5，v/v/v/v）用于 HSCCC 实践，从青霉的粗乙酸乙酯提取物中一步分离出三种化合物；从 100mg 粗提物中获得 6.1mg 化合物 1、5.6mg 化合物 2 和 3.8mg 化合物 3，固定相保留率为 75%。然后将这些化合物鉴定为大黄素甲醚、大黄酚和大黄素。通过测试对α-葡萄糖苷酶活性的抑制作用和分子对接模拟，验证了目标化合物在青霉次级代谢产物中的活性。结果表明，大黄素、大黄酚和大黄素甲醚对α-葡萄糖苷酶活性具有显著的抑制作用。这项工作证实了 HSCCC 在分离复杂提取物中的化合物方面的有效性，并为青霉的进一步研究和应用提供了参考。
