Determination of 1-hydroxypyrene in children urine using column-switching liquid chromatography and fluorescence detection

Abstract

This study developed an acid hydrolysis method instead of using enzyme extraction, equipped with column-switching system for the pretreatment of samples, in the determination of 1-hydroxypyrene in the urine from children and pyrene in airborne particulates. We collected both types of samples from areas near a petrochemical industry and rural areas as reference. Samples were first treated with acid hydrolysis and followed by solvent extraction prior to being injected into the separation system for the determination with high performance liquid chromatography and fluorescence. A column-switching system was on-line with a C 18 separation column to remove matrix interference and obtain a stable baseline of the chromatogram. The eluent used to separate the 1-hydroxypyrene was 60% (v/v) aqueous acetonitrile solution. A fluorescence detector was used to monitor 1-hydroxypyrene at λ ex =348 nm and λ em =388 nm , and pyrene at λ ex =331 nm and λ em =390 nm . Both calibration graphs were linear with very good correlation coefficients ( r>0.999) and the detection limits were ca. 2 pg (5 ng/l). Results showed that there was a significant association between 1-hydroxypyrene levels in urine specimens and pyrene levels in airborne particulate samples ( r=0.68, P<0.05). The average levels of pyrene in the particulates (0.18 versus 0.09 ng/m 3) and of 1-hydroxypyrene in urine specimens (155.9 versus 110.2 ng/g creatinine) were higher for the petrochemical area than for the rural area. This method is stable and sensitive for measuring polycyclic aromatic hydrocarbons in environmental samples.