Planta Med 2014; 80 - SL35
DOI: 10.1055/s-0034-1394523

Optimized MS-based isolation strategy for rapid targeted purification of antifungal compounds

A Azzollini 1, J Zhang 2, Q Favre-Godal 1, E Ferreira-Queiroz 1, S Wang 2, D Guillarme 1, P Fan 2, H Lou 2, JL Wolfender 1
  • 1School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, 30 Quai Ansermet CH-1211 Geneva 4, Switzerland
  • 2Department of Natural Products Chemistry, Key Lab of Chemical Biology of the Ministry of Education, School of Pharmaceutical Sciences, Shandong University, No. 44 West Wenhua Road, Jinan 250012, People's Republic of China

The targeted purification of bioactive molecules from complex extracts is of prime importance in the field of drug discovery from natural sources. In this regard, Mass Spectrometry (MS) detection is a key tool enabling the monitoring of specific features for precise fractionation. To improve the isolation process efficiency of antifungal natural products (NPs), an LC-MS-based purification strategy was developed. First, the chromatographic separation of the crude extract was optimized by application of a linear gradient at the analytical scale in HPLC-UV-MS. At-line microfractionation, followed by agar overlay bioautography using a Candida albicans hypersusceptible strain, was performed to identify the bioactive fractions [1]. The gradient was then geometrically transferred from the analytical to the preparative scale using gradient transfer rules based on the calibration of both chromatographic systems [2]. Finally, an MS-triggered isolation of the localized bioactive molecules was realized with a Flash chromatographic system coupled, via splitter, to a single quadrupole mass spectrometer (PuriFlash® – MS). This isolation strategy was applied for the MS-direct purification of the antifungal compound Diplophyllolide A from the Chinese liverwort Chiloscyphus Polyanthos (L.) Cord. The bioactive molecule was directly isolated in large amount, without issued related to MS saturation, thanks to the optimization of the MS splitting geometry. This rational LC-MS-based methodology has high potential for the rapid isolation of compounds of interest identified by MS-based metabolomics and for the efficient purification of bioactive natural products that lack of UV chromophore.

Keywords: MS-direct purification, antifungal compounds, Flash Chromatography, Chiloscyphus Polyanthos.

References:

[1] Q. Favre-Godal, S. Dorsaz, E.F. Queiroz, C. Conan et al., Phytochemistry, Accepted.

[2] D. Guillarme, D.T.T. Nguyen, S. Rudaz, J.-L. Veuthey, Eur. J. Pharma. Biopharma. 2008, 68, 430.