Determination of nitrotyrosine by HPLC-ECD and its application.
Determination of nitrotyrosine by HPLC-ECD and its application.
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DOI：
10.1292/jvms.64.401
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发表时间：
2002-05
期刊：
影响因子：
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通讯作者：
N. Ishida;T. Hasegawa;K. Mukai;Masanobu Watanabe;H. Nishino
中科院分区：
文献类型：
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作者：
N. Ishida;T. Hasegawa;K. Mukai;Masanobu Watanabe;H. Nishino
3-硝基酪氨酸是酪氨酸硝化的产物，是氧化损伤的有用指标。我们修改了以前报道的HPLC-电化学检测（ECD）方法：具体地说，通过型多孔碳电极被用作还原电极，而不是汞-金汞齐电极，因为后者的响应随时间而变化。反相HPLC和电化学检测器的组合通过-800 mV还原电位，随后在+250 mV氧化电位下，允许测量3-硝基酪氨酸。该测定的检测限小于10 fmol。在腹腔内给予脂多糖（LPS）的小鼠中，血浆3-硝基酪氨酸水平升高，对应于LPS剂量。结果表明，改进的HPLC-ECD法可作为生物3-硝基酪氨酸的特异、灵敏的检测方法，可用于临床。
3-nitrotyrosine, a product of tyrosine nitration, is a useful indicator of oxidative damage. We modified the previously reported HPLC-electrochemical detection (ECD) method: specifically, a through-type porous carbon electrode was used as a reducing electrode instead of the mercury-gold amalgam electrode, because the response of the latter changes over time. A combination of reverse-phase HPLC and electrochemical detector passed through -800 mV reduction potential and subsequently under +250 mV oxidation potential allows measurement of 3-nitrotyrosine. The detection limit of this assay was less than 10 fmol. In mice to which lipopolysaccharide (LPS) was administered intraperitoneally, plasma 3-nitrotyrosine levels were elevated, corresponding to LPS dosage. These findings suggest that the improved HPLC-ECD method can be used as a specific and sensitive assay of biological 3-nitrotyrosine and can be applied clinically.