Benzo[a]pyrene – Determination of 3-hydroxybenzo[a]pyrene in urine by LC-MS/MS

Biomonitoring Method – Translation of the German version from 2023

Gerhard Scherer¹ (Method development)
Heinz-Werner Hagedorn¹ (Method development)
Hans-Wolfgang Hoppe² (External verification)
Paulina Dadanski² (External verification)
Albrecht Seidel³ (External verification)
  Thomas Göen⁴ (Head of the working group “Analyses in Biological Materials” of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area, Deutsche Forschungsgemeinschaft)
  Andrea Hartwig⁵ (Chair of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area, Deutsche Forschungsgemeinschaft)
  MAK Commission<sup>6

1</sup> ABF Analytisch-Biologisches Forschungslabor GmbH, Semmelweisstraße 5, 82152 Planegg, Germany
² MVZ Medizinisches Labor Bremen GmbH, Haferwende 12, 28357 Bremen, Germany
³ Biochemical Institute for Environmental carcinogens Prof. Dr. Gernot Grimmer-Foundation, Lurup 4, 22927 Großhansdorf, Germany
⁴ Friedrich-Alexander-Universität Erlangen-Nürnberg, Institute and Outpatient Clinic of Occupational, Social, and Environmental Medicine, Henkestraße 9–11, 91054 Erlangen, Germany
⁵ Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, Building 50.41, 76131 Karlsruhe, Germany
⁶ Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area, Deutsche Forschungsgemeinschaft, Kennedyallee 40, 53175 Bonn, Germany

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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