Determination of urinary 8-hydroxy-2'-deoxyguanosine, trans, trans-muconic acid, and S-phenylmercapturic acid by liquid chromatography-mass spectrometry

Abstract

To establish a method for simultaneously determining the urinary concentrations of 8-hydroxy-2'-deoxyguanosine (8-OHdG), trans, trans-muconic acid (tt-MA), and S-phenylmercapturic acid (S-PMA) in subjects exposed to benzene. After being purified by a solid-phase extraction column, the urine samples were transferred to a liquid chromatography-mass spectrometry system, and the concentrations of 8-OHdG, tt-MA, and S-PMA were determined by external standard method. A C18 reversed-phase column was used as the chromatographic column, and methanol/acidic ammonium formate solution was used as the mobile phase for gradient elution. The mass spectrometer was operated in a multi-reaction monitoring mode. For tt-MA, the calibration curves were linear in the range of 10-1000 µg/L, and the recovery rates were over 90% (relative standard deviation (RSD) < 3%) at spiked levels of 50 µg/L and 500 µg/L. For S-PMA and 8-OHdG, the calibration curves were linear in the range of 1-100 µg/L, and the recovery rates were over 85% (RSD < 5%) at spiked levels of 5 µg/L and 50 µg/L. This determination method meets the requirement of Biological materials- of monitoring-Guide of development (WS/T 68-1996) and can be used for simultaneous determination of 8-OHdG, tt-MA, and S-PMA in urine.