Journal
TOXICOLOGY LETTERS
Volume 181, Issue 1, Pages 29-37Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.toxlet.2008.06.867
Keywords
3-Bromotyrosine; Human urine; Liquid chromatography mass spectrometry; 3-Nitrotyrosine; Urinary protein
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Funding
- National Science Council of Taiwan [NSC 94-2113-M-194-014, NSC 95-2119-M-194-004]
- National Chung Cheng University
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Nitration and bromination of proteins, giving rise to the respective 3-nitrotyrosine (3NT) and 3-bromotyrosine (3BT), are implicated in asthma, allergic inflammatory disorders, and cancer. We have developed an isotope dilution liquid chromatography electrospray ionization tandem mass spectrometry (LC/MS/MS) assay for simultaneous analysis of protein-bound 3NT and 3BT in human urine. The detection limits (S/N=3) were 10 pg (44 fmol) for 3NT and 5.0 pg (19 fmol) for 3BT injected on-column. The average levels of protein-bound 3NT and 3BT in 23 healthy individuals were 9.7 +/- 11.0 (mean +/- S.D.) in 10(5) tyrosine and 4.4 +/- 3.9 (mean +/- S.D.) in 10(3) tyrosine, respectively, using this highly sensitive LC/MS/MS under the selective reaction monitoring mode. Furthermore, the levels of urinary 3NT and 3BT show a statistically significant correlation (R-2 = 0.55, p = 0.0065, n = 23). The high specificity and accuracy of this LC/MS/MS method render it a valuable tool in measurement of 3NT and 3BrT in the human urinary protein as promising noninvasive biomarkers for protein tyrosine nitration and bromination in vivo. (C) 2008 Elsevier Ireland Ltd. All rights reserved.
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