Musk xylene (MX) is frequently used as a fragrance in commercial toiletries. Biotransformation of MX into 4-amino-MX (4-AMX) and 2-amino-MX (2-AMX) metabolites in rainbow trout haemoglobin (Hb) has been described. The dose–response relationship and toxicokinetics of the metabolites as adducts in the Hb were determined by gas chromatography (GC)–electron capture negative chemical ionization (NCI)–mass spectrometry (MS), and GC–electron ionization (EI)–MS/MS, using selected ion monitoring (SIM). The trout were subjected to a single exposure of 0.010, 0.030, 0.10, and/or 0.30mgMX/g of fish. Hb samples were collected from exposed and control fish, and analysed subsequent to exposure at intervals of 24, 72, and 168h. Alkaline hydrolysis released 4-AMX and 2-AMX metabolites from the Hb, and the solutes were extracted into n -hexane. The extracts were preconcentrated and analysed. The presence of the metabolites in the Hb extracts was confirmed based on agreement of similar mass spectral features from NCI/MS and EI-MS/MS spectra, and retention times of the metabolites with standards. The NCI/MS results were used for dose–response and toxicokinetics measurements. For dose–response, the concentrations of adducts of the metabolites increased with dosage, and a maximum adduct formation was observed at 0.10mgg −1 , beyond which it decreased. The average concentrations of 4-AMX and 2-AMX at a dosage of 0.10mgg −1 were 700 and 7.4ngg −1 , respectively. For toxicokinetics, the concentration of the metabolites in the Hb reached a maximum in the 3 day sample after administration of MX. Further elimination of the metabolites exhibited kinetics with a half-life estimated to be 1–2 days, assuming first-order kinetics. Quantitations were made based on an internal standard and a calibration plot. In control samples, non-hydrolysed Hb, and reagent blank extracts, the metabolites were not detected. The limits of detection for 4-AMX and 2-AMX in the Hb were approximately 1.7 and 1.4µgL −1 , respectively, based on a signal-to-noise ratio of 3 with NCI/MS.
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Document Type: Research Article
Department of Chemistry and Biochemistry, Baylor University, PO Box 97348, Waco, TX 76798, USA
US Environmental Protection Agency, Quality Assurance Laboratory, 944 East Harmon Ave, Las Vegas, NV 89119, USA
August 20, 2006
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