Planta Med 1991; 57(2): 142-148
DOI: 10.1055/s-2006-960051

© Georg Thieme Verlag Stuttgart · New York

Thermospray Liquid Chromatography/Mass Spectrometry (TSP LC/MS) Analysis of the Alkaloids from Cinchona in vitro Cultures

C. Giroud1 , T. van der Leer1 , R. van der Heijden1 , R. Verpoorte1 , C. E. M. Heeremans2 , W. M. A. Niessen2 , J. vander Greef2
  • 1Biotechnology Delft Leiden, Project-group Plant Cell Biotechnology, Division of Pharmacognosy, Center for Bio-Pharmaceutical Sciences, Gorlaeus Laboratories. P.O.Box 9502, NL-2300 RA Leiden, The Netherlands
  • 2Division of Analytical Chemistry, Center for Bio-Pharmaceutical Sciences, Gorlaeus Laboratories, P.O. Box 9502, 2300 RA Leiden, The Netherlands
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Publication History


Publication Date:
05 January 2007 (online)


The alkaloids from Cinchona ledgeriana shoot cultures and from Cinchona robusta shoot cultures and a compact globular structure (CGS) culture were analyzed by thermospray liquid chromatography/mass spectrometry (TSP LC/MS). Because of the relative stability of the alkaloids under TSP discharge ionization conditions, a protonated molecule was observed in the mass spectra with hardly any fragmentation. When the reference compounds were available, the knowledge of the molecular mass and of the retention time was sufficient to identify most of the alkaloids. HPLC with UV photodiode-array detection complemented LC/MS perfectly by providing information about the aromatic part of the alkaloids (structure and substitution pattern). New alkaloids detected in Cinchona in vitro cultures were 5-methoxytryptamine and corynantheal. In order to determine whether 5-methoxytryptamine was a precursor of the methoxylated quinolines, this indole was incubated with secologanin and several Cinchona robusta crude protein extracts. Under all conditions tested, the coupling of 5-methoxytryptamine with secologanin remained unsuccessful. Only tryptamine condensed with secologanin to yield strictosidine. These results indicate that Cinchona cells are able to methoxylate simple indoles like tryptamine and that 5-methoxytryptamine is very likely not used for the subsequent biosynthesis of the methoxylated quinolines.