Zhejiang Academy of Forestry, Hangzhou, 310023, Zhejiang Province China.
Zhejiang Academy of Forestry, Hangzhou, 310023, Zhejiang Province China.
J Chromatogr A. 2021 Jul 19;1649:462221. doi: 10.1016/j.chroma.2021.462221. Epub 2021 May 8.
Improved closed-loop recycling counter-current chromatography (CLR CCC) with a two-phase solvent system composed of n-hexane-acetonitrile (1:1, v/v) was developed for separation, purification and preparation of cyclosporin D from the crude extract of fungus Hypoxylon Spp. (sj18). 28 mg cyclosporin D was successfully purified from 300 mg crude extract sample. The purity was 95.2% after five cycles, determined by HPLC. The structure of cyclosporin D was identified and assigned by H NMR, C NMR and mass spectrometric analyses. In addition, in the study, we show an interesting phenomenon that cyclosporin D can be prepared by the conventional CCC in n-hexane-ethyl acetate-methanol-water solvent system (2.5:1:2.5:1, v/v/v/v), and can also be prepared by the improved closed-loop recycling CCC in n-hexane-acetonitrile solvent system (1:1, v/v), but the efficiency of preparation varies greatly.
开发了一种由正己烷-乙腈（1:1，v/v）组成的两相溶剂体系的改进型闭路循环逆流色谱（CLR CCC），用于从真菌 Hypoxylon Spp.（sj18）的粗提物中分离、纯化和制备环孢菌素 D。从 300mg 粗提物样品中成功纯化出 28mg 环孢菌素 D。经 HPLC 测定，纯度为 95.2%。通过 1H NMR、13C NMR 和质谱分析鉴定并分配了环孢菌素 D 的结构。此外，在研究中，我们展示了一个有趣的现象，即环孢菌素 D 可以通过传统的 CCC 在正己烷-乙酸乙酯-甲醇-水溶剂系统（2.5:1:2.5:1，v/v/v/v）中制备，也可以通过改进的闭路循环回收 CCC 在正己烷-乙腈溶剂系统（1:1，v/v）中制备，但制备效率差异很大。
