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Methods Mol Biol. 2012;828:255-70. doi: 10.1007/978-1-61779-445-2_20.

Measurement of 3-nitro-tyrosine in human plasma and urine by gas chromatography-tandem mass spectrometry.

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Institute of Clinical Pharmacology, Hannover Medical School, Hannover, Germany.


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.

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