Sensitive determination of nitrotyrosine in human plasma by isocratic high-performance liquid chromatography |
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Authors: | Yoshinori Kamisaki Kouichirou Wada Kentaro Nakamoto Yosuke Kishimoto Masayuki Kitano Tadao Itoh |
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Institution: | aDepartment of Clinical Pharmacology, Faculty of Medicine, Tottori University, 86 Nishimachi, Yonago 683, Japan;bSecond Department of Internal Medicine, Faculty of Medicine, Tottori University, 86, Nishimachi, Yonago 683, Japan |
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Abstract: | A highly sensitive and simple isocratic high-performance liquid chromatography 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 α-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. |
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Keywords: | Nitrotyrosine |
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