Optimization of Sample Preparation for 1-Hydroxypyrene as a Major Biomarker of Exposure to PAHs Prior to HPLC

Urinary 1-hydroxypuren (1-ohp) is commonly used as a major metabolite and biological indicator of the overall exposureto polycyclic aromatic hydrocarbons (pahs). for evaluation of human exposure to such compounds, biological monitoringis an essential process, in which, preparation of samples is one of the most time-consuming and error-prone aspects prior tochromatographic techniques. in this study, non classic form of liquid-liquid extraction (lle) was optimized with regard tosolvent type, solvent volume, extraction temperature, mixing type, and mixing duration. through the extraction process, amild temperature was used to keep the compound of interest as stable as possible. in this study, a high performance liquidchromatography, using reverse-phase column was used. the isocratic run was done at a constant flow rate of 0.8 ml/min,the mobile phase was methanol/water and a fluorescence detector was used, setting at 242 nm and 388 nm. at the developedconditions, the extraction recovery was exceeded 87.3%, achieving detection limit of 0.2 µg/l. the factors were evaluatedstatically and the procedure was validated with three different pools of spiked urine samples and showed a goodreproducibility over six consecutive days as well as experiments. it was concluded that, this optimized method couldsimplify sample preparation for trace residue analysis of pahs metabolites.