Planta Med 2016; 82(S 01): S1-S381
DOI: 10.1055/s-0036-1596260
Abstracts
Georg Thieme Verlag KG Stuttgart · New York

Flash chromatography MS-targeted isolation of natural products under normal phase conditions

D Righi
1   School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, 30 Quai Ansermet CH-1211 Geneva 4, Switzerland
,
A Azzollini
1   School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, 30 Quai Ansermet CH-1211 Geneva 4, Switzerland
,
EF Queiroz
1   School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, 30 Quai Ansermet CH-1211 Geneva 4, Switzerland
,
JL Wolfender
1   School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, 30 Quai Ansermet CH-1211 Geneva 4, Switzerland
› Author Affiliations
Further Information

Publication History

Publication Date:
14 December 2016 (online)

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    Reversed phase liquid chromatography (RP-LC) is widely used for the metabolite profiling of complex natural extracts and is now more and more used for targeted MS isolation of biomarkers. Normal phase chromatography (NP-LC) is well suited for the purification of lipophilic secondary metabolites, also offering some advantages compared to RP, like low operating pressures and cheaper stationary phases. The complementary usage of both of stationary phases and MS detection for metabolite purification at the preparative scale using generic separation methods has been investigated on an innovative Flash chromatography system integrating MS and ELSD detection (PuriFlash®- MS).

    A mixture of representative natural product standards was chosen and analysed under both normal phase/reverse phase conditions. All parameters were carefully optimized for both separation and detection (gradient system, split rate, flow rate, temperature, inj. volume, column length, ionization source parameters). A special care was taken to find MS ionization and splitting conditions that provide good detection and preclude source contamination. The HPLC analytical gradient was transferred to flash chromatography following a geometric gradient transfer method after calibration of the chromatographic systems [1]. MS, in complement to UV detection, enabled the monitoring of NPs with weak and strong chromophores and the selectivity of MS was of great help for a precise collection of partially co-eluting compounds. APCI-MS detection with optimized splitting and post-column elution of appropriate solvent was found robust and well-suited for purifications in both NP and RP modes. This strategy allows an efficient and rational targeted isolation of tens to hundreds mg of compounds for further structural identification or bioactivity characterization studies.

    Acknowledgements: Interchim (Monluçon, France) for loan of instrument and technical support.

    Keywords: Flash chromatography, APCI-MS, analytical method transfer, normal phase, reverse phase

    References:

    [1] Davy Guillarme, Dao T.T. Nguyen, Serge Rudaz, Jean-Luc Veuthey. Method transfer for fast liquid chromatography in pharmaceutical analysis: Application to short columns packed with small particle. Part II: Gradient experiments. Eur J Pharma Biopharma 2008; 68: 430 – 440


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    No conflict of interest has been declared by the author(s).