Benzene is an important occupational and environmental contaminant, naturally present in petroleum and as by-product in the steel industry. Toxicological studies showed pronounced myelotoxic action, causing leukemic and others blood cells disorders. Assessing of benzene exposure is performed by biomarkers as trans, trans-muconic acid (AttM) and S-phenylmercapturic acid (S-PMA) in urine. Due to specificity of S-PMA, this biomarker has been proposed to asses lower levels of benzene in air. The aim of this study was to validate an analytical method for the quantification of S-PMA by High-Performance Liquid Chromatography with fluorometric detector. The development of an analytical method of S-PMA in urine was carried out by solid phase extraction (SPE) using C-18 phase. The eluated were submitted to water bath at 75 °C and nitrogen to analyte concentration, followed by alkaline hydrolysis and derivatization with monobromobimane. The chromatography conditions were reverse phase C-18 column (240 mm, 4 mm and 5 μm) at 35 °C; acetonitrile and 0.5% acetic acid (50:50) as mobile phase with a flow of 0.8 mL/min. The limits of detection and quantification were 0.22 μg/L and 0.68 μg/L, respectively. The linearity was verified by simple linear regression, and the method exhibited good linearity in the range of 10–100 μg/L. There was no matrix effect for S-PMA using concentrations of 40, 60, 80 and 100 μg/L. The intra- and interassay precision showed coefficient of variation of less than 10% and the recovery ranged from 83.4 to 102.8% with an average of 94.4%. The stability of S-PMA in urine stored at −20 °C was of seven weeks. The conclusion is that this method presents satisfactory results per their figures of merit. This proposed method for determining urinary S-PMA showed adequate sensitivity for assessment of occupational and environmental exposure to benzene using S-PMA as biomarker of exposure.

苯是一种重要的职业和环境污染物，天然存在于石油中，是钢铁行业的副产品。毒理学研究显示出明显的骨髓毒性作用，引起白血病和其他血细胞疾病。尿中苯的暴露程度可以通过生物标志物如反式，反式-粘康酸（AttM）和S-苯基巯基酸（S -PMA）进行评估。由于S -PMA的特异性，已提出使用该生物标记物来评估空气中较低的苯含量。这项研究的目的是验证一种定量分析S的分析方法。-PMA采用高效液相色谱仪，带有荧光检测器。通过使用C-18相的固相萃取（SPE）进行了尿液中S -PMA分析方法的开发。将洗脱液置于75°C的水浴中，在氮气中浓缩至分析物浓度，然后进行碱水解并用单溴二苯胺衍生化。色谱条件为在35°C下使用反相C-18柱（240 mm，4 mm和5μm）；乙腈和0.5％乙酸（50:50）作为流动相，流速为0.8毫升/分钟。检测限和定量限分别为0.22μg/ L和0.68μg/ L。通过简单的线性回归验证了线性，该方法在10–100μg/ L的范围内表现出良好的线性。S没有基质效应-PMA使用40、60、80和100μg/ L的浓度。批内和批间精密度的变异系数均小于10％，回收率介于83.4％至102.8％之间，平均值为94.4％。S -PMA在-20°C的尿液中的稳定性为7周。结论是，该方法根据其品质因数显示出令人满意的结果。该提议的测定尿中S -PMA的方法显示出足够的敏感性，可以使用S -PMA作为暴露的生物标志物来评估职业和环境中苯的暴露。