Preparative Scale MS Guided Isolation of Bioactive Compounds Using High Resolution Flash Chromatography: Antifungals from Chiloscyphus polyanthos as a Case Study.
In natural product research, the efficient purification of molecules from large amounts of complex extracts is a key element. In this regard, an integrative strategy for efficient MS guided isolation of antifungal compounds has been developed. First, off line HPLC antifungal activity based profiling and HPLC PDA MS profiling were used to localize the compounds of interest on the analytical scale. Then, the analytical gradient was geometrically transferred to the flash chromatographic level. Finally, an MS triggered isolation of the localized bioactive molecules was realized using high resolution flash chromatographic columns (15 um spherical particles) coupled to a single quadrupole mass spectrometer via a splitter system. This isolation strategy was applied for the MS targeted purification of antifungal principles from the liverwort Chiloscyphus polyanthos. This rational methodology has high potential for the targeted large scale purification of bioactive compounds, avoiding the need to repeat a given bioassay at each isolation step. Seven sesquiterpene lactones were isolated, of which five were found to be bioactive and one was reported as a new compound. The absolute configuration of some compounds was established for the first time by electronic circular dichroism spectroscopy.
Planta medica