Determination of Nitrotyrosine by HPLC-ECD and Its Application

Journal of Veterinary Medical Science

Hirohito NISHINO²  Kazutaka MUKAI¹  Nobushige ISHIDA¹  Masanobu WATANABE²  Telhisa HASEGAWA¹ 

[1] Equine Research Institute, Japan Racing Association;Eicom Corporation

关键词: HPLC-ECD;    LPS;    3-nitrotyrosine;    peroxynitrite;   

DOI  :  10.1292/jvms.64.401

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(13)Cited-By(7)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.

【 授权许可】

Unknown