Improved HPLC method for analysis of 1-hydroxypyrene in human urine specimens of cigarette smokers

A method for the rapid and sensitive analysis of 1-hydroxypyrene in human urine was improved by high performance liquid chromatography (HPLC) with fluorescence detection. The limit of detection decreased approximately threefold when triethylamine was added to the extracts prior to the chromatographic determination. The sensitivity was increased mainly due to triethylamine blockage of the secondary-retention effects. The analysis time was approximately 20 min per run. This method was applied to the analysis of 1-hydroxypyrene in 125 urine specimens collected from cigarette smokers who had non-occupational exposure to polycyclic aromatic hydrocarbons. The average concentrations of urinary 1-hydroxypyrene were 0.04 micromol/mol creatinine in non-smokers, 0.20 micromol/mol creatinine in light smokers, 0.46 micromol/mol creatinine in medium smokers, and 1.16 micromol/mol creatinine in heavy cigarette smokers. The levels of urinary 1-hydroxypyrene correlated strongly with cigarette intake (r2 = 0.93, P < 0.001).