<div class="csl-bib-body">
<div class="csl-entry">da Silva Amaral, L., Rodrigues-Filha, E., Kubicek, C. P., Herwig, C., Marchetti-Deschmann, M., & Allmaier, G. (2018). Optimization of sample preparation for intact cell mass spectrometry (matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometry) of endophytic Xylaria. <i>Rapid Communications in Mass Spectrometry</i>, <i>32</i>(10), 815–823. https://doi.org/10.1002/rcm.8098</div>
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dc.identifier.issn
0951-4198
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/144989
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dc.description.abstract
Rationale
Although the fruiting-body of the fungi of the genus Xylaria shows a great variety of morphological characteristics, their mycelial forms are always very similar, imposing difficulties for their identification. Intact cell mass spectrometry (ICMS) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) can be a fast and reliable strategy to support the differentiation/identification of Xylaria species in those cases where fruit-bodies are not available.
Methods
Many experimental parameters such as sample preparation and culture media are crucial for filamentous fungi analysis by MALDI-TOFMS. For the purposes of this study, we used four matrices (CHCA, DHB, FA and SA) with five different concentrations (0.1, 0.3, 0.5, 1.0 and 2.5%) of TFA in the matrix, the influence of six different culture media (solid and liquid), and three mycelium peptide/protein extraction protocols (acid, basic and thymol-supported solution) to optimize the sample preparation of the endophytic fungus X. arbuscula.
Results
It was observed that sinapinic acid (30 mg/mL) dissolved in acetonitrile/0.1% TFA and PDA were the best matrix solution and culture medium, respectively, for the ICMS of X. arbuscula. The formic acid and ammonium bicarbonate (AB) protocols provided similar mass spectra; however, a higher number of peaks were observed using AB extraction. Mass spectra obtained from different thymol-containing solutions (EtOH/aqueous 0.1% TFA and ACN/aqueous 0.1% TFA) show increasing peak abundances at m/z 3000–6500.
Conclusions
X. arbuscula could be analyzed by ICMS. However, an extraction step was required to provide suitable MALDI mass spectra. Formic acid-, AB- and thymol-containing solutions were demonstrated to be good cocktails for the extraction of peptide/protein biomarkers from these fungi.
en
dc.language.iso
en
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dc.publisher
WILEY
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dc.relation.ispartof
Rapid Communications in Mass Spectrometry
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dc.subject
Spectroscopy
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dc.subject
Analytical Chemistry
en
dc.subject
Organic Chemistry
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dc.title
Optimization of sample preparation for intact cell mass spectrometry (matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometry) of endophytic Xylaria
en
dc.type
Artikel
de
dc.type
Article
en
dc.description.startpage
815
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dc.description.endpage
823
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dc.type.category
Original Research Article
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tuw.container.volume
32
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tuw.container.issue
10
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tuw.journal.peerreviewed
true
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tuw.peerreviewed
true
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wb.publication.intCoWork
International Co-publication
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tuw.researchTopic.id
M6
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tuw.researchTopic.name
Biological and Bioactive Materials
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tuw.researchTopic.value
100
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dcterms.isPartOf.title
Rapid Communications in Mass Spectrometry
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tuw.publication.orgunit
E166-05 - Forschungsbereich Biochemische Technologie