An Integrated Chromatographic Strategy for the Large-Scale Extraction of Ergosterol from Tulasnellaceae sp.

Abstract

A reliable chromatographic strategy is crucial for the extraction of target compounds from natural sources as it is related to the preparation efficiency, as well as the purity of the compounds. In this study, medium-pressure normal-phase liquid chromatography and high-pressure reverse-phase liquid chromatography were combined to prepare and purify ergosterol from Tulasnellaceae sp. of Gymnadenia orchidis. First, Tulasnellaceae sp. was extracted three times (2.0 L and 2 h each time) with ethyl acetate, and the 6.0 L of extract solution was concentrated under reduced pressure to yield 2.2 g of crude sample. Then, the crude sample was pretreated utilizing silica gel medium-pressure liquid chromatography to enrich the target ingredient (586.0 mg). Finally, high-pressure reversed-phase liquid chromatography was used to purify the target compound, and the compound was characterized as ergosterol (purity > 95%) using spectral data. Overall, the simple and reproducible integrated chromatographic strategy developed in this study has the potential for the large-scale purification of steroids for laboratory and even industrial research. To the best of our knowledge, this is also the first report of ergosterol in Tulasnellaceae sp.