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    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 1H NMR, 13C 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. Copyright © 2021. Published by Elsevier B.V.


    Hepeng Li, Feiying Zhang, Qunying Jin, Tangjun Zhu. Preparative separation and purification of Cyclosporin D from fungus Hypoxylon Spp. by improved closed-loop recycling counter-current chromatography. Journal of chromatography. A. 2021 Jul 19;1649:462221

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    PMID: 34034103

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