Assay of 3‐Nitrotyrosine in Tissues and Body Fluids by Liquid Chromatography with Tandem Mass Spectrometric Detection
Abstract
3-Nitrotyrosine (3-NT) is a marker of protein nitration in physiological systems. It is present as 3-nitrotyrosine residues in proteins of tissue, extracellular matrix, plasma, and other body fluids and food. It is also present in body fluids and some beverages as free nitrotyrosine and is excreted in urine with the major urinary metabolite 3-nitro-4-hydroxyphenylacetic acid. Quantitation of 3-nitrotyrosine requires tandem mass spectrometry for specific detection. The method developed to determine 3-nitrotyrosine (along with protein glycation and oxidation adducts in a quantitative screening assay) by liquid chromatography with tandem mass spectrometric detection is described. The 3-NT residue contents of plasma protein, hemoglobin, lipoproteins, and cerebrospinal fluid protein and the concentrations of free 3-nitrotyrosine in plasma, urine, and cerebrospinal fluid are given. Changes of 3-nitrotyrosine residue and free 3-nitrotyrosine in diabetes, cirrhosis, acute and chronic renal failure, and neurological disorders, including Alzheimer's disease, are presented and compared with independent estimates.
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Several limitations of ELISA for 3-NT have been highlighted in the literature, including low sensitivity, minor specificity of applied antibodies, susceptibility to errors, and missing detection of free 3-NT.
“several limitations have been highlighted in the literature, such as low sensitivity, minor specificity of applied antibodies, the susceptibility to errors, or missing detection of free 3-NT”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [197]
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References 68 · Cited by 32
- Authors
- Rabbani, N., & Thornalley, P. J.
- Affiliations
- University Hospital Coventry (GB)
- Venue
- Methods in Enzymology
- Publisher
- Elsevier
- Published
- 2008
- Volume
- 440
- Pages
- 337-359
- ISSN
- 0076-6879 (ISSN-L)
- OpenAlex
- W1889537242
- Cited by
- 32
- References
- 68
- Citations by year
- 2024: 2 · 2022: 4 · 2021: 4 · 2017: 2 · 2015: 2 · 2014: 1 · 2013: 2 · 2012: 3
- FWCI
- 5.17
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Methods in enzymology on CD-ROM/Methods in enzymology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Wellcome Trust
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Advanced Glycation End Products research
- Keywords
- nitrotyrosine, protein nitration, plasma proteins, hemoglobin, lipoproteins, cerebrospinal fluid, urine, diabetes mellitus, liver cirrhosis, renal failure, Alzheimer's disease, oxidative stress biomarkers
- MeSH terms
- Animals, Body Fluids, Chromatography, Liquid, Humans, Organ Specificity, Tyrosine, Tandem Mass Spectrometry
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