Analytical and Bioanalytical Chemistry

, Volume 408, Issue 8, pp 1997–2008 | Cite as

Analysis of tetrahydroxylated benzo[a]pyrene isomers in hair as biomarkers of exposure to benzo[a]pyrene

  • Nathalie Grova
  • Emilie M. Hardy
  • Pauline Meyer
  • Brice M. R. Appenzeller
Paper in Forefront
First Online:
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Revised:
Accepted:

Abstract

A first gas chromatography–tandem mass spectrometry (GC–MS/MS) method was designed for analysis of four tetrahydroxylated benzo[a]pyrene metabolites (benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol, benzo[a]pyrene-r-7,t-8,t-9,t-10-tetrahydrotetrol, benzo[a]pyrene-r-7,t-8,c-9,c-10-tetrahydrotetrol, and benzo[a]pyrene-r-7,t-8,c-9,t-10-tetrahydrotetrol) in hair. Hair powder extract was submitted to liquid–solid extraction, followed by C18 solid-phase purification. The analytes were derivatized with use of N-methyl-N-(trimethylsilyl)trifluoroacetamide and then analyzed by GC–MS/MS in negative chemical ionization mode. The calibration curve was linear from the limit of quantification (LOQ) to 20 pg/mg in hair. The coefficient of determination of the calibration curve was more than 0.975 for all the analytes investigated. The LOQs ranged from 0.075 to 0.2 pg/mg in hair. The method was afterward applied to the analysis of hair of 16 rats randomly allocated to experimental groups receiving 16 polycyclic aromatic hydrocarbons solubilized in oil at 0 or 0.8 mg/kg body weight by oral administration three times per week for 90 days. The analysis of monohydroxylated and dihydroxylated benzo[a]pyrenes was conducted in parallel by GC–MS/MS on the same samples. All tetrahydroxylated benzo[a]pyrene isomers were detected in hair samples collected from rats exposed to polycyclic aromatic hydrocarbons. Benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol, the most abundant isomer in hair of treated rats, was also the principal isomer released in DNA adduct hydrolysis in humans. Moreover, the benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol concentrations in hair were significantly greater than those of 2-hydroxybenzo[a]pyrene, 1-hydroxybenzo[a]pyrene, 7-hydroxybenzo[a]pyrene, and 4-hydroxybenzo[a]pyrene and similar to those of 9-hydroxybenzo[a]pyrene and 3-hydroxybenzo[a]pyrene. The method was also sufficiently sensitive to monitor environmental levels of exposure because two hair specimens in the eight analyzed from smokers were above the LOQ for benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol and benzo[a]pyrene-r-7,t-8,c-9,t-10-tetrahydrotetrol. This study therefore demonstrated that tetrahydroxylated benzo[a]pyrenes in hair might be a useful biomarker for the assessment of both the general population and occupationally exposed workers.

Graphical Abstract

GC-NCI-MS/MS analysis of tetrahydroxylated benzo[a]pyrene metabolites in hair of rat under controlled exposure to benzo[a]pyrene (10 mg/kg, 28 days)

Keywords

Polycyclic aromatic hydrocarbons Tetrahydroxylated benzo[a]pyrenes Hair analysis Rat model Gas chromatography–tandem mass spectrometry 
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Notes

Acknowledgments

This work was supported by the Luxembourg Ministère de l’Enseignement Supérieur et de la Recherche (MESR), the Agence de l’Environnement et de la Maîtrise de l’Énergie (ADEME), and the Agence Nationale de Sécurité Sanitaire de l’Alimentation, de l’Environnement du Travail (ANSES). We thank Henri Schroeder, who allowed the performance of animal experiments with benzo[a]pyrene alone in Nancy (URAFPA-INRA UC340). We are most grateful to Marie France Schoën, Chantal Courtois, Stephanie Sallai, and Anaïs Oudin for their technical assistance.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest

Supplementary material

216_2016_9338_MOESM1_ESM.pdf (103 kb)
ESM 1 (PDF 102 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Nathalie Grova
    • 1
  • Emilie M. Hardy
    • 1
  • Pauline Meyer
    • 1
  • Brice M. R. Appenzeller
    • 1
  1. 1.Human Biomonitoring Research UnitLuxembourg Institute of Health, House of BiohealthEsch-sur-AlzetteLuxembourg

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