Mykotoxine – Bestimmung von Deoxynivalenol und Deepoxydeoxynivalenol in Urin mittels LC-MS/MS
Biomonitoring-Methode
Marion Berger1 (Methodenentwicklung)Lennart Marske1 (Methodenentwicklung)
Bernhard Monien2 (Methodenprüfung)
Solveigh Siodlaczek2 (Methodenprüfung)
Thomas Göen3 (Leitung der Arbeitsgruppe „Analysen in biologischem Material“ der Ständigen Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft)
Andrea Hartwig4 (Vorsitz der Ständigen Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft)
MAK Commission5
1 Bundesanstalt für Arbeitsschutz und Arbeitsmedizin (BAuA), Fachbereich 4 – Gefahrstoffe und biologische Arbeitsstoffe, Gruppe 4.2 – Medizinischer Arbeitsschutz, Biomonitoring, Nöldnerstraße 40/42, 10317 Berlin, Deutschland
2 Bundesinstitut für Risikobewertung, Fachgruppe 54 – Abt. Lebensmittelsicherheit, Max-Dohrn-Straße 8–10, 10589 Berlin, Deutschland
3 Friedrich-Alexander-Universität Erlangen-Nürnberg, Institut und Poliklinik für Arbeits-, Sozial- und Umweltmedizin, Henkestraße 9–11, 91054 Erlangen, Deutschland
4 Institut für Angewandte Biowissenschaften, Abteilung Lebensmittelchemie und Toxikologie, Karlsruher Institut für Technologie (KIT), Adenauerring 20a, Geb. 50.41, 76131 Karlsruhe, Deutschland
5 Ständige Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft, Kennedyallee 40, 53175 Bonn, Deutschland
Abstract
The working group “Analyses in Biological Materials” of the German Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area (MAK Commission) developed and verified the presented biomonitoring method. The aim of this method is the selective and sensitive quantitation of deoxynivalenol (DON; free DON plus glucuronides not otherwise specified) and its metabolite deepoxydeoxynivalenol (DOM-1) in urine. After enzymatic hydrolysis of the urine sample and purification of the analytes on an immunoaffinity column, followed by preconcentration of the eluates under a stream of nitrogen, determination is carried out by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Calibration is performed with comparative standards prepared in urine and treated analogously to the samples to be analysed. DON is quantified using an internal standard (ISTD; 13C15-DON), whereas DOM-1 is quantified without the use of an ISTD. Good precision data with standard deviations below 9% for DON and below 6% for DOM-1, as well as good accuracy data with mean relative recoveries in the range of 93–114% for DON and 97–103% for DOM-1, show that the method provides reliable and accurate analytical results. The method is both selective and sensitive, and has a limit of quantitation of 0.179 μg/l for DON and of 0.26 μg/l for DOM-1. Due to rapid renal excretion, the method is primarily suitable for analysing acute exposure which occurred only hours prior to sampling.
