Analytical Methods for Secondary Metabolite Detection

  • Judith Taibon
  • Hermann StrasserEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1477)


The entomopathogenic fungi Metarhizium brunneum, Beauveria bassiana, and B. brongniartii are widely applied as biological pest control agent in OECD countries. Consequently, their use has to be flanked by a risk management approach, which includes the need to monitor the fate of their relevant toxic metabolites. There are still data gaps claimed by regulatory authorities pending on their identification and quantification of relevant toxins or secondary metabolites. In this chapter, analytical methods are presented allowing the qualitative and quantitative analysis of the relevant toxic B. brongniartii metabolite oosporein and the three M. brunneum relevant destruxin (dtx) derivatives dtx A, dtx B, and dtx E.

Key words

Secondary metabolite Toxin Oosporein Destruxin Persistence Risk assessment Analyte monitoring Analytical tool HPLC–DAD HPLC–DAD–QTOF–MS/MS 



This research has been supported by the European Community’s Seventh Framework Programme grant (FP7_ENV.2011.3.1.9-1 ECO-INNOVATION, INBIOSOIL, Grant Agreement No. 282767). We grateful acknowledge the contribution from PD Dr. Christoph Seger, Dr. Sonja Sturm and Prof. Dr. Hermann Stuppner (all University Innsbruck) who provided their expertise within this project.


  1. 1.
    Seger C, Sturm S, Längle T, Wimmer W, Stuppner H, Strasser H (2005) Development of a sensitive high-performance liquid chromatography-diode array detection assay for the detection and quantification of the Beauveria metabolite oosporein from submerged culture broth and bio-control formulations. J Agric Food Chem 53:1364–1369CrossRefPubMedGoogle Scholar
  2. 2.
    Seger C, Längle T, Pernfuss B, Stuppner H, Strasser H (2005) High-performance liquid chromatography-diode array detection assay for the detection and quantification of the Beauveria metabolite oosporein from potato tubers. J Chromatogr A 1092:254–257CrossRefPubMedGoogle Scholar
  3. 3.
    Seger C, Eberhart K, Sturm S, Strasser H, Stuppner H (2006) Apolar chromatography on Sephadex LH-20 combined with high-speed counter-current chromatography. High yield strategy for structurally closely related analytes-Destruxin derivatives from Metarhizium anisopliae as a case study. J Chromatogr A 1117:67–73CrossRefPubMedGoogle Scholar
  4. 4.
    Seger C, Sturm S, Stuppner H, Butt TM, Strasser H (2004) Combination of a new sample preparation strategy with an accelerated high-performance liquid chromatography assay with photodiode array and mass spectrometric detection for the determination of destruxins from Metarhizium anisopliae culture broth. J Chromatogr A 1061:35–43CrossRefPubMedGoogle Scholar
  5. 5.
    Jegorov A, Havlícek V, Sedmera P (1998) Rapid screening of destruxins by liquid chromatography/mass spectrometry. J Mass Spectrom 33:274–280CrossRefPubMedGoogle Scholar
  6. 6.
    Kershaw MJ, Moorhouse ER, Bateman R (1999) The role of destruxins in the pathogenicity of Metarhizium anisopliae for three species of insect. J Invertebr Pathol 74:213–223CrossRefPubMedGoogle Scholar
  7. 7.
    Loutelier C, Cherton JC, Lange C, Traris M, Vey A (1996) Studies on the dynamics of the production of destruxins by Metarhizium anisopliae direct high-performance liquid chromatographic and fast atom bombardment mass spectrometric analysis correlated with biological activity tests. J Chromatogr A 738:181–189CrossRefGoogle Scholar
  8. 8.
    Potterat O, Wagner K, Haag H (2000) Liquid chromatography–electrospray time-of-flight mass spectrometry for on-line accurate mass determination and identification of cyclodepsipeptides in a crude extract of the fungus Metarrhizium anisopliae. J Chromatogr A 872:85–90CrossRefPubMedGoogle Scholar
  9. 9.
    Wang C, Skrobek A, Butt TM (2003) Concurrence of losing a chromosome and the ability to produce destruxins in a mutant of Metarhizium anisopliae. FEMS Microbiol Lett 226:373–378CrossRefPubMedGoogle Scholar
  10. 10.
    Wang C, Skrobek A, Butt TM (2004) Investigations on the destruxin production of the entomopathogenic fungus Metarhizium anisopliae. J Invertebr Pathol 85:168–174CrossRefPubMedGoogle Scholar
  11. 11.
    Morais RP, Lira SP, Seleghim MHR, Berlinck RGS (2010) A method for dextruxin analysis by HPLC-PDA-ELSD-MS. J Braz Chem Soc 21:2262–2271CrossRefGoogle Scholar
  12. 12.
    Hsiao Y, Ko J (2001) Determination of destruxins, cyclic peptide toxins, produced by different strains of Metarhizium anisopliae and their mutants induced by ethyl methane sulfonate and ultraviolet using HPLC method. Toxicon 39:837–841CrossRefPubMedGoogle Scholar
  13. 13.
    Taibon J, Sturm S, Seger C, Parth M, Strasser H, Stuppner H (2014) Development of a fast and selective UHPLC-DAD-QTOF-MS/MS method for the qualitative and quantitative assessment of destruxin profiles. Anal Bioanal Chem 406:7623–7632CrossRefPubMedGoogle Scholar
  14. 14.
    Liu C, Huang S, Tzeng Y (2004) Analysis of destruxins produced from Metarhizium anisopliae by capillary electrophoresis. J Chromatogr Sci 42:140–144CrossRefPubMedGoogle Scholar
  15. 15.
    Taibon J, Sturm S, Seger C, Werth M, Strasser H, Stuppner H (2015) Supercritical fluid chromatography as an alternative tool for the qualitative and quantitative analysis of Metarhizium brunneum metabolites from culture broth. Planta Med. doi: 10.1055/s-0035-1557823 Google Scholar
  16. 16.
    Newmeyer D (1990) Filtering small quantities of conidial suspensions to remove mycelial fragments. Fungal Genet Newslett 37:27Google Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute of Pharmacy, Department of Pharmacognosy, CCB – Centrum of Chemistry and BiomedicineUniversity of InnsbruckInnsbruckAustria
  2. 2.Institute of MicrobiologyUniversity of InnsbruckInnsbruckAustria

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