Abstract
Reaction of reactive nitrogen species (RNS), such as peroxynitrite and nitryl chloride with soluble tyrosine and tyrosine residues in proteins produces soluble 3-nitro-tyrosine and 3-nitro-tyrosino-proteins, respectively. Regular proteolysis of 3-nitro-tyrosino-proteins yields soluble 3-nitro-tyrosine. 3-Nitro-tyrosine circulates in plasma and is excreted in the urine. Both circulating and excretory 3-nitro-tyrosine are considered suitable biomarkers of nitrative stress. Tandem mass spectrometry coupled with gas chromatography (GC–MS/MS) or liquid chromatography (LC–MS/MS) is one of the most reliable analytical techniques to determine 3-nitro-tyrosine. Here, we describe protocols for the quantitative determination of soluble 3-nitro-tyrosine in human plasma and urine by GC–MS/MS.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Tsikas D, and Caidahl K (2005) Recent methodological advances in the mass spectrometric analysis of free and protein-associated 3-nitro-tyrosine in human plasma. J Chromatogr B 814:1–9
Ryberg H, and Caidahl K (2007) Chromatographic and mass spectrometric methods for quantitative determination of 3-nitro-tyrosine in biological samples and their application to human samples. J Chromatogr B 851:160–171
Tsikas D (2010) Analytical methods for 3-nitro-tyrosine quantification in biological samples: the unique role of tandem mass spectrometry. Amino Acids, DOI 10.1007/s00726-010-0604-5.
Tsikas D et al (2005) Determination of 3-nitro-tyrosine in human urine at the basal state by gas chromatography-tandem mass spectrometry and evaluation of the excretion after oral intake. J Chromatogr B 827:146–156
Kato Y et al (2009) Quantification of modified tyrosines in healthy and diabetic human urine using liquid chromatography/tandem mass spectrometry. J Clin Biochem Nutr 44:67–78
Tsikas D et al (2010) Evidence by GC–MS of ex vivo nitrite and nitrate formation from air nitrogen oxides (NOx) in human plasma, serum and urine samples. Anal Biochem 397:126–128
Schwedhelm E, Sandmann J, Tsikas D (1998) Synthesis of 3-nitro-L-(2H3)tyrosine for use as an internal standard for quantification of 3-nitro-L-tyrosine by gas chromatography-mass spectrometry. J Labelled Cpds Radiopharm 41:773–780
Söderling AS et al (2003) A derivatization assay using gas chromatography/negative chemical ionization tandem mass spectrometry to quantify 3-nitro-tyrosine in human plasma. J Mass Spectrom 38:1187–1196
Schwedhelm E et al (1999) Gas chromatographic-tandem mass spectrometric quantification of free 3-nitro-tyrosine in human plasma at the basal state. Anal Biochem 276:195–203
Tsikas D et al (2003) Accurate quantification of basal plasma levels of 3-nitro-tyrosine and 3-nitrotyrosinoalbumin by gas chromatography-tandem mass spectrometry. J Chromatogr B 784:77–90
Tsikas D (2008) A critical review and discussion of analytical methods in the L-arginine/nitric oxide area of basic and clinical research. Anal Biochem 379: 139–163
Keimer R et al (2003) Lack of oxidative stress during sustained therapy with isosorbide dinitrate and pentaerythrityl tetranitrate in healthy humans: a randomized, double-blind crossover study. J Cardiovasc Pharmacol 41:284–292
Tsikas D (2006) Quantitative determination of free and protein-associated 3-nitro-tyrosine and S-nitrosothiols in the circulation by mass spectrometry and other methologies: a critical review and discussion from the analytical and review point of view. In: Dalle-Donne I, Scaloni A, Butterfield DA (eds) Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases, John Wiley & Sons Inc., Hoboken
Thum T et al (2007) Endothelial nitric oxide synthase uncoupling impairs endothelial progenitor cell mobilization and function in diabetes. Diabetes 56: 666–674
Magné J et al (2009) Rapeseed protein in a high-fat mixed meal alleviates postprandial systemic and vascular oxidative stress and prevents vascular endothelial dysfunction in healthy rats. J Nutr 139:1660–1666
Pham VV, Stichtenoth DO, and Tsikas D (2009) Nitrite correlates with 3-nitro-tyrosine but not with the F(2)-isoprostane 15(S)-8-iso-PGF(2alpha) in urine of rheumatic patients. Nitric Oxide 21:210–215
Göen T et al (2005) Sensitive and accurate analyses of free 3-nitro-tyrosine in exhaled breath condensate by LC–MS/MS. J Chromatogr B 826:261–266
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tsikas, D., Mitschke, A., Gutzki, FM. (2012). Measurement of 3-Nitro-Tyrosine in Human Plasma and Urine by Gas Chromatography-Tandem Mass Spectrometry. In: Alterman, M., Hunziker, P. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 828. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-445-2_20
Download citation
DOI: https://doi.org/10.1007/978-1-61779-445-2_20
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-444-5
Online ISBN: 978-1-61779-445-2
eBook Packages: Springer Protocols