Benzo[a]pyren – Bestimmung von 3-Hydroxybenzo[a]pyren in Urin mittels LC-MS/MS

Biomonitoring-Methode

Gerhard Scherer¹ (Methodenentwicklung)
Heinz-Werner Hagedorn¹ (Methodenentwicklung)
Hans-Wolfgang Hoppe² (Methodenprüfung)
Paulina Dadanski² (Methodenprüfung)
Albrecht Seidel³ (Methodenprüfung)
  Thomas Göen⁴ (Leitung der Arbeitsgruppe „Analysen in biologischem Material“ der Ständigen Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft)
  Andrea Hartwig⁵ (Vorsitz der Ständigen Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft)
  MAK Commission<sup>6

1</sup> ABF Analytisch-Biologisches Forschungslabor GmbH, Semmelweisstraße 5, 82152 Planegg, Deutschland
² MVZ Medizinisches Labor Bremen GmbH, Haferwende 12, 28357 Bremen, Deutschland
³ Biochemisches Institut für Umweltcarcinogene Prof. Dr. Gernot Grimmer Stiftung, Lurup 4, 22927 Großhansdorf, Deutschland
⁴ Friedrich-Alexander-Universität Erlangen-Nürnberg, Institut und Poliklinik für Arbeits-, Sozial- und Umweltmedizin, Henkestraße 9–11, 91054 Erlangen, Deutschland
⁵ Institut für Angewandte Biowissenschaften, Abteilung Lebensmittelchemie und Toxikologie, Karlsruher Institut für Technologie (KIT), Adenauerring 20a, Geb. 50.41, 76131 Karlsruhe, Deutschland
⁶ Ständige Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe, Deutsche Forschungsgemeinschaft, Kennedyallee 40, 53175 Bonn, Deutschland

Abstract

The working group “Analyses in Biological Materials” of the German Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area developed and verified the presented biomonitoring method. Benzo[a]pyrene (B[a]P) is a polycyclic aromatic hydrocarbon and is classified as carcinogenic to humans by the Commission. B[a]P originates, for example, from coal tar or from the incomplete combustion of organic material. The general population is primarily exposed through cigarette smoke, food, and ambient air. The aim of this work was to develop a selective method for the determination of 3-hydroxybenzo[a]pyrene (3-OH-B[a]P) in urine. The method is characterised by the use of glucuronidated standards, ascorbic acid as an antioxidant, and by the conversion of the analyte into a derivative which allows for sensitive measurement. The procedure has been comprehensively validated, and the reliability data have been confirmed by replication and verification of the procedure in a second, independent laboratory. For the determination of 3-OH-B[a]P, an internal standard is added to the urine samples, which undergo solid-phase extraction prior to analysis by liquid chromatography-tandem mass spectrometry. Quantitative evaluation is carried out via external calibration using pooled urine from persons with no known exposure to B[a]P. The good precision and accuracy data show that the method provides reliable and accurate analytical results. The method is both selective and sensitive, and the limit of detection of 17 pg/l urine is sufficient to determine both occupational exposure and exposure to B[a]P in the general population.

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