title: HPLC-fast scanning fluorimetric detection determination of risk exposure to polycyclic aromatics hydrocarbons biomarkers in human urine.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Analytical Chemistry, University of Extremadura, 06006 Badajoz, Spain.
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Luque-Uría
authors.affiliation: Department of Analytical Chemistry & Research Institute on Water, Climate Change & Sustainability (IACYS), University of Extremadura, 06006 Badajoz, Spain.
authors.forename: Anunciación
authors.initials: A
authors.lastname: Espinosa-Mansilla
authors.affiliation: Department of Analytical Chemistry & Research Institute on Water, Climate Change & Sustainability (IACYS), University of Extremadura, 06006 Badajoz, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Durán-Merás
mesh_terms: D015415:Biomarkers / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004784:Environmental Monitoring / Q000379:methods*; D006801:Humans; D011084:Polycyclic Aromatic Hydrocarbons / Q000652:urine*; D012307:Risk Factors; D012907:Smoking
chemical_list: D015415:Biomarkers; D011084:Polycyclic Aromatic Hydrocarbons
keywords: HPLC; biomarkers; hydroxyl-polycyclic aromatic hydrocarbons; spectrofluorimetry; urinary metabolites