On-line identification of the antifungal constituents of Erythrina vogelii by liquid chromatography with tandem mass spectrometry, ultraviolet absorbance detection and nuclear magnetic resonance spectrometry combined with liquid chromatographic micro-fractionation
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Abstract
In our continuing search for new antifungal agents of plant origin, the investigation
of Erythrina vogelii Hook. f. (Leguminosae), a plant used in the traditional medicine
of Ivory Coast to treat various infectious ailments, was undertaken. In order to rapidly
identify the active principles, the crude extract was analysed by low-flow LC-1H nuclear
magnetic resonance spectrometry (NMR) which gave a sensitive detection of all the
main peaks. LC microfractionation was performed just after LC-NMR detection and all
peaks collected were submitted to antifungal bioautography assays against Cladosporium
cucumerinum. By this means, the antifungal activity could be efficiently linked to
three of the LC peaks. In order to obtain complementary on-line structural information
for all peaks of interest, high-resolution LC-MS-MS together with LC-UV with post-column
addition of UV shifts reagents was undertaken on the crude extract. This chemical
screening strategy with integrated antifungal bioassays has permitted the on-line
identification of numerous constituents and has given useful information for an efficient
peak-guided isolation procedure.