title: Reverse phase high performance liquid chromatographic method development based on ultravioletvisible detector for the analysis of 1-hydroxypyrene (PAH biomarker) in human urine.
abstract: OBJECTIVE
  1-hydroxypyrene is an important biomarker of exposure to polycyclic aromatic hydrocarbons (PAHs), which appears in the urine of exposed human subjects. In developing countries, where advanced instruments are not available, the importance of this biomarker demands convenient and sensitive methods for determination purposes. This study aimed at developing a methodology to quantify 1-hydroxypyrene (a biomarker of PAHs exposure) based on the UV-visible detector in the reverse phase high pressure liquid chromatography (HPLC).
  
  
  METHODS
  A 20 μl injection of sample was used for manual injection into the HPLC Shimadzu, equipped with the SPD-20 A UV-visible detector, the LC-20AT pump and the DGU-20A5 degasser. The C-18 column was used for the purpose of the analysis.
  
  
  RESULTS
  The method showed a good linearity (the range: R2= 0.979-0.989), and high detectability up to the nmol level. The average retention was 6.37, with the accuracy of 2%, and the percentage of recovery remained 108%. The overall performance of this method was comparable (in terms of detection sensitivity) and relatively better than previously reported studies using the HPLC system equipped with the UV-detector.
  
  
  CONCLUSIONS
  This method is suitable and reliable for the detection/quantification of the 1-OHP in human urine samples, using the UV-detector, however, it is less sensitive as compared to the results of a florescence detector.
authors.affiliation: Quaid-i-Azam University, Islamabad, Pakistan (Faculty of Biological Sciences, Department of Environmental Sciences, Environmental Biology and Ecotoxicology Laboratory).
authors.forename: Atif
authors.initials: A
authors.lastname: Kamal
authors.affiliation: Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi, Pakistan (Institute of Biochemistry).
authors.forename: Mohammad
authors.initials: M
authors.lastname: Gulfraz
authors.affiliation: Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi, Pakistan (Institute of Biochemistry).
authors.forename: Mohammad Asad
authors.initials: MA
authors.lastname: Anwar
authors.affiliation: Quaid-i-Azam University, Islamabad, Pakistan (Faculty of Biological Sciences, Department of Environmental Sciences, Environmental Biology and Ecotoxicology Laboratory). r_n_malik2000@yahoo.co.uk.
authors.forename: Riffat Naseem
authors.initials: RN
authors.lastname: Malik
mesh_terms: D015415:Biomarkers / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D004781:Environmental Exposure; D006801:Humans; D011084:Polycyclic Aromatic Hydrocarbons; D011721:Pyrenes / Q000652:urine*; D014466:Ultraviolet Rays
chemical_list: D015415:Biomarkers; D011084:Polycyclic Aromatic Hydrocarbons; D011721:Pyrenes; C033146:1-hydroxypyrene
keywords: 1-OHP; 1-hydroxypyrene; PAHs; Pakistan; biomarkers of exposure; high pressure liquid chromatography