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Simultaneous quantification of enterotoxins tilimycin and tilivalline in biological matrices using HPLC high resolution ESMS2 based on isotopically 15N-labeled internal standards.
Glabonjat, Ronald A; Kitsera, Maksym; Unterhauser, Katrin; Lembacher-Fadum, Christian; Högenauer, Christoph; Raber, Georg; Breinbauer, Rolf; Zechner, Ellen L.
Afiliação
  • Glabonjat RA; Institute of Chemistry, NAWI Graz, University of Graz, A-8010, Graz, Austria.
  • Kitsera M; Institute of Molecular Biosciences, University of Graz, A-8010, Graz, Austria.
  • Unterhauser K; Institute of Molecular Biosciences, University of Graz, A-8010, Graz, Austria.
  • Lembacher-Fadum C; Institute of Organic Chemistry, Graz University of Technology, A-8010, Graz, Austria.
  • Högenauer C; BioTechMed, Graz, Austria; Division of Gastroenterology and Hepatology, Department of Internal Medicine, Medical University of Graz, A-8036, Graz, Austria.
  • Raber G; Institute of Chemistry, NAWI Graz, University of Graz, A-8010, Graz, Austria.
  • Breinbauer R; Institute of Organic Chemistry, Graz University of Technology, A-8010, Graz, Austria; BioTechMed, Graz, Austria.
  • Zechner EL; Institute of Molecular Biosciences, University of Graz, A-8010, Graz, Austria; BioTechMed, Graz, Austria; Field of Excellence BioHealth - University of Graz, Graz, Austria. Electronic address: ellen.zechner@uni-graz.at.
Talanta ; 222: 121677, 2021 Jan 15.
Article em En | MEDLINE | ID: mdl-33167283
ABSTRACT
Non-ribosomal peptides are one class of bacterial metabolites formed by gut microbiota. Intestinal resident Klebsiella oxytoca produces two pyrrolobenzodiazepines, tilivalline and tilimycin, via the same nonribosomal biosynthesis platform. These molecules cause human disease by genotoxic and tubulin inhibitory activities resulting in apoptosis of the intestinal epithelium, loss of barrier integrity and ultimately colitis. Here we report a fast, reliable, HPLC-HR-ESMS2 method for quantifying simultaneously the bacterial enterotoxins tilimycin and tilivalline in complex biological matrices. We synthesized and applied stable isotopically labeled internal standards for precise quantification of the metabolites. Sample preparation was optimized using clinical and laboratory specimens including serum, colonic fluid and stool. The developed method overcame the disadvantage of low selectivity by applying high resolution mass spectrometry in MS2 mode. High sensitivity and low interference from matrices were achieved and validated. We show that the approach is suitable for detection and quantification of the enterotoxic metabolites produced in vivo, in infected human or animal hosts, and in bacterial culture in vitro.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Benzodiazepinonas / Enterotoxinas Tipo de estudo: Guideline Limite: Animals / Humans Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Benzodiazepinonas / Enterotoxinas Tipo de estudo: Guideline Limite: Animals / Humans Idioma: En Ano de publicação: 2021 Tipo de documento: Article