Sensitive determination of nitrotyrosine in human plasma by isocratic high-performance liquid chromatography.
A highly sensitive and simple isocratic high-performance liquid chromatographic method was developed for determination of 3-nitrotyrosine in human plasma with precolumn derivatization with 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole. The precision of the method was satisfactory (coefficient of variation 4.8%), and the detection limit was established at 0.1 pmol of 3-nitrotyrosine allowing the determination at the level of 6 pmol/ml in human plasma. The recoveries of 3-nitrotyrosine and alpha-methyltyrosine, an internal standard, were 89.3 +/- 7.1 and 85.7 +/- 7.6%, respectively. The 3-nitrotyrosine level was 31 +/- 6 pmol/ml (mean +/- S.D., n = 9) in plasma from healthy volunteers. Since 3-nitrotyrosine is a stable product of peroxynitrite, an oxidant formed by a reaction of nitric oxide and superoxide radicals, the measurement of its plasma concentration may be useful as a marker of nitric oxide-dependent oxidative damage.
Kamisaki
Y
Y
Department of Clinical Pharmacology, Faculty of Medicine, Tottori University, Yonago, Japan.
Wada
K
K
Nakamoto
K
K
Kishimoto
Y
Y
Kitano
M
M
Itoh
T
T
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives* / Q000737:chemistry; D015415:Biomarkers / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry
D015415:Biomarkers; D005456:Fluorescent Dyes; C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; C002744:3-nitrotyrosine; D014443:Tyrosine; D009327:4-Chloro-7-nitrobenzofurazan