Hplc-Fast Scanning Fluorimetric Detection Determination of Risk Exposure to Polycyclic Aromatics Hydrocarbons Biomarkers in Human Urine

Abstract

Aim: An HPLC method for the determination of 2-hydroxyfluorene (2-OHF), various hydroxyphenanthrene metabolites (1-, 2-, 3-, 4- and 9-hydroxyphenanthrene, OHPhs), 1-hydroxypyrene (1-OHPy) and 3-hydroxybenzo[a]pyrene (3-OHB[a]Py) in human urine, has been developed using fast scanning fluorimetric detection and gradient elution mode. Materials & methods: All reagents were of analytical grade. Standard solutions were prepared separately, by exact weighing or dilution with ultrapure acetonitrile, and were stored at 4 ºC in darkness. The standard addition method was used for the analysis of urine samples. Results: In the optimized conditions, 2- and 3-hydroxyphenanthrene, and 1- and 9-hydroxyphenanthrene metabolites eluted at the same retention time; however, all other hydroxy-polycyclic aromatic hydrocarbons were well resolved. Multi-emission detection allows us to monitor each metabolite at its most sensitivity emission wavelength. Detection limits ranged between 0.9 and 4.26 ng ml^-1. Conclusion: Fortified urine samples of nonexposure and nonsmoker volunteers, previous precipitation step with acetonitrile, were used to test the proposed method. The obtained results confirm the goodness of the method.

Keywords::

• biomarkers
• HPLC
• hydroxyl-polycyclic aromatic hydrocarbons
• spectrofluorimetry
• urinary metabolites

Financial & competing interests disclosure

The authors are grateful to the Ministerio de Economia y Competitividad of Spain (Project CTQ2014-52309-P) and the Gobierno de Extremadura (GR15090 Research Group FQM003) both co-financed by the European FEDER funds, for finance of this work. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

For investigations involving human subjects, informed consent has been obtained from the participants involved. Samples have been voluntarily provided by the researchers.