Quantitative Determination of Free and Protein‐Associated 3‐Nitrotyrosine and S ‐Nitrosothiols in the Circulation by Mass Spectrometry and Other Methodologies: A Critical Review and Discussion from the Analytical and Review Point of View

Mass spectrometry and 3-nitrotyrosine: Strategies, controversies, and our current perspective

“…The array of physicochemical and immunological approaches that have been applied to the analysis of soluble 3nitrotyrosine (Tables 2 and 3) and protein-associated 3-nitrotyrosine (Table 4) determinations has been reviewed from several perspectives (Herce-Pagliai, Kotecha, & Shuker, 1998;Duncan, 2003;Aulak et al, 2004;Tsikas & Caidahl, 2005;Tsikas, 2006Tsikas, , 2010aTsikas, , 2012Ryberg & Caidahl, 2007;Bigelow & Qian, 2008;Murray et al, 2008;Nuriel et al, 2008;Rabbani & Thornalley, 2008;Sharov et al, 2008;Abello et al, 2009;Toda et al, 2010;Spickett & Pitt, 2012;Tsikas, Mitschke, & Gutzki, 2012). Human plasma 3nitrotyrosine concentrations determined by these methods range from 0.6 nM (LC-MS/MS; Svatikova et al, 2004) to 5,300 nM (HPLC-UV; Fatouros et al, 2004), whereas EBC levels range from 0.04 nM (GC-MS/MS; Lärstad et al, 2005 and LC-MS/ MS; Göen et al, 2005) to 35 nM (HPLC-FL; Balint et al, 2001).…”

“…With respect to tyrosine in the membrane, however, nitration but not oxidation of tyrosine by peroxynitrite has been reported to prevail (Zhang et al, ). In humans, the reaction of RNS with tyrosine and thiols leads to very low yields of 3‐nitrotyrosine and S ‐nitrosothiols in biological fluids, respectively (Tsikas, Schwedhelm, & Frölich, ; Tsikas, ; Giustarini et al, ; Ryberg & Caidahl, ). Interestingly, the reaction of RNS with other biomolecules such as unsaturated fatty acids also yields pM/nM concentrations of nitrofatty acids in biological fluids in vitro (Jain et al, ) and in vivo (Tsikas et al, ).…”

“…The arrows indicate in which direction the formation of 3‐nitrotyrosine (red colored) or GSSG (brown colored) changes upon addition (+) of CO 2 . This figure was reconstructed by using previously reported data (Tsikas, ).…”