Mass spectrometry guided purification for efficient isolation of natural products at semi- and preparative scale

In natural product research the isolation of compounds from crude extracts is a key element. In this respect, the increase of the purification process' efficiency, by improvement of instrumental and methodological approaches, is a crucial point. To tackle this issue, a two-steps chromatographic strategy was developed. First, the separation of target molecules from a crude extract was optimized by application of a linear gradient at the analytical scale. Second, the gradient was geometrically transferred to semi-preparative HPLC by a gradient transfer method based on the calibration of the chromatographic systems (measurement of dwell volume and extra column volume) [1]. UV as well as MS monitoring were performed for a comprehensive detection of the various compounds present within the mixture. MS detection proved to be an important tool, not only for the purification of compounds that cannot be detected by UV, but also for an efficient mass spectrometry guided purification of specific compounds in crude extracts. In particular, a single quadrupole mass spectrometer coupled to a semi-preparative chromatographic system (PuriFlash® – MS) was found a promising tool to increase the efficiency of the isolation of constituents of interest. This MS-guided separation of compounds in complex mixtures represents a powerful strategy, not only for the isolation of molecules present in bioactive fractions, but also for the rapid purification of biomarkers identified by UHPLC-MS metabolomics and dereplication process.

References:

[1] Davy Guillarme, Dao T.T. Nguyen, Serge Rudaz, Jean-Luc Veuthey, Eur. J. Pharma. Biopharma. 2008, 68, 430.