Journal
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
Volume 23, Issue 6, Pages 877-884Publisher
WILEY
DOI: 10.1002/rcm.3949
Keywords
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Funding
- Austrian Federal Ministry of Agriculture, Forestry, Environment and Water Management
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Unambiguous identification of mycotoxin-producing fungal species as Fusarium is of great relevance to agriculture and the food-producing industry as well as in medicine. Protein profiles of intact fungal spores, such as Penicillium, Aspergillus and Trichoderma, derived from matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were shown to provide a rapid and straightforward method for species identification and characterization. In this study, we applied this approach to five different Fusarium spp. strains which are known to affect the growth of different grain plants. To obtain a suitable MALDI matrix system and sample preparation method, thin-layer, dried-droplet and sandwich methods and several MALDI matrices, namely CHCA, DHB, FA, SA and TRAP dissolved in various solvent mixtures (organic solvents such as ACN, MeOH, EtOH and iPrOH and for the aqueous phase water and 0.1% TFA), were evaluated in terms of mass spectrometric pattern and signal intensities. The most significant peptide/protein profiles were obtained with 10 mg ferulic acid (FA) in 1 mL ACN/0.1% TFA (7:3, v/v) used as matrix system. Mixing the spores with the matrix solution directly on the MALDI target (dried-droplet technique) resulted in an evenly distributed spores/matrix crystal layer, yielding highly reproducible peptide/protein profiles from the spore surfaces. Numerous abundant ions throughout the investigated m/z range (m/z 1500-15000) could be detected. Differences in the obtained mass spectral patterns allowed the differentiation of spores of various Fusarium species. Copyright (C) 2009 John Wiley & Sons, Ltd.
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