The Analysis of Pyrrolizidine Alkaloids in Jacobaea vulgaris; a Comparison of Extraction and Detection Methods

L. Joosten, P.P.J. Mulder, K. Vrieling, M.R. van der Veen, P.G.L. Klinkhamer

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    Introduction – Pyrrolizidine alkaloids (PAs) serve an important function in plant defence. Objective – To compare different extraction methods and detection techniques, namely gas chromatography with nitrogen phosphorus detection (GC-NPD) and liquid chromatography tandem mass spectrometry (LC-MS/MS) with quadrupole analysers for analysing PAs in Jacobaea vulgaris. Methodology – Both formic acid and sulfuric acid were tested for PA extraction from dry plant material. For GC-NPD, reduction is required to transform PA N-oxides into tertiary amines. Zinc and sodium metabisulfite were compared as reducing agents. Results – The lowest PA concentration measured with GC-NPD was approximately 0.03¿mg/g and with LC-MS/MS 0.002¿mg/g. The detection of major PAs by both techniques was comparable but a number of minor PAs were not detected by GC-NPD. With the LC-MS/MS procedure higher concentrations were found in plant extracts, indicating that losses may have occurred during the sample preparation for the GC-NPD method. Zinc proved a more effective reducing agent than sodium metabisulfite. The sample preparation for LC-MS/MS analysis using formic acid extraction without any reduction and purification steps is far less complex and less time consuming compared to GC-NPD analysis with sulfuric acid extraction and PA N-oxide reduction with zinc and purification. Conclusions – In terms of sensitivity and discrimination, formic acid extraction in combination with LC-MS/MS detection is the method of choice for analysing PAs (both free and N-oxides forms) in plant material
    Original languageEnglish
    Pages (from-to)197-204
    JournalPhytochemical Analysis
    Issue number2
    Publication statusPublished - 2010


    • senecio-jacobaea
    • mass-spectrometry
    • n-oxides
    • insects
    • root


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