An online automatic sample cleanup system for the quantitative detection of the benzene exposure biomarker S-phenylmercapturic acid in human urine by electrospray ionization tandem mass spectrometry

An online automatic sample cleanup system was developed for use with electrospray ionization tandem mass spectrometry (ESI-MS-MS) for the quantitative detection of the benzene exposure biomarker S-phenylmercapturic acid (S-PMA) in human urine. The sample clean-up system was constructed with an autosampling device, a reversed-phase C18 trap cartridge, a two-position switching valve, and controlling computer software and hardware. The sample cleanup system was interfaced directly with the ESI source of a triple-stage-quadrupole MS using multiple reaction monitoring of negative product ions derived from S-PMA and the internal standard as the detection mode. The calibration curve was linear using human urine spiked at concentrations from 0.25 to 100 mg/L S-PMA (R² = 0.997). The detection limit of the analytical system for neat S-PMA standard solution was 0.04 μ0g/L, whereas the detection limit was estimated to be lower than 0.35 μg/L for a urine matrix containing trace amounts of S-PMA. Without tedious manual sample cleanup procedures, the analytical system is fully automatic and therefore useful for high-throughput urinary S-PMA determination. With the selectivity and the sensitivity provided by MS-MS detection, the analytical system can be used for high-throughput and accurate determination of S-PMA levels in human urinary samples as a biomarker for benzene exposure.