3-Nitrotyrosine quantification methods: Current concepts and future challenges
Open accessAbstract
Background Measurement of 3-nitrotyrosine (3-NT) in biological samples can be used as a biomarker of nitrosative stress, since it is very stable and suitable for analysis. Increased 3-NT levels in biological samples have been associated with several physiological and pathological conditions. Different methods have been described for the detection and quantification of this molecule, such as (i) immunological methods; (ii) liquid chromatography, namely high-pressure liquid chromatography (HPLC)-based methods that use ultraviolet-visible (UV/VIS) absorption, electrochemical (ECD) and diode array (DAD) detection, liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS); (iii) gas chromatography, such as gas chromatography-mass spectrometry (GC-MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS). Methods A literature review on nitrosative stress, protein nitration, as well as 3-NT quantification methods was carried out. Results This review covers the different methods for analysis of 3-NT that have been developed during the last years as well as the latest advances in this field. Overall, all methods present positive and negative aspects, although it is clear that chromatography-based methods present good sensitivity and specificity. Regarding this, GC-based methods exhibit the highest sensibility in the quantification of 3-NT, although it requires a prior time consuming derivatization step. Conversely, HPLC does not require such derivatization step, despite being not as accurate as GC. Conclusion It becomes clear that all the methods described during this literature review, although accurate for 3-NT quantification, need to be improved regarding both sensitivity and specificity. Moreover, optimization of the protocols that have been described is clearly needed.
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Protein carbonyls, 3-nitrotyrosine and methionine sulfoxide are biomarkers of protein oxidation commonly analyzed via immunochemical methods.
“Protein carbonyls, 3-NT and methionine sulfoxide are biomarkers of protein oxidation which are commonly analyzed via immunochemical methods [75,82,92].”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [92]
Section 5, “Quantification of 3-nitrotyrosine in biological samples” (page not provided), says that a large part of 3-NT research has used antibody-based methods. The paper supports that part of the sentence, but the cited passages do not establish that protein carbonyls or methionine sulfoxide are commonly analyzed by immunochemical methods. (LLM, conf=0.88)
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Jones, D. P. (2006) · Antioxidants & Redox Signaling · doi:10.1089/ars.2006.8.1865 · Cited by 1,726 · see citing works
Rose, S., Melnyk, S., Pavliv, O., Bai, S., Nick, T. G., Frye, R. E., & James, S. J. (2012) · Translational Psychiatry · doi:10.1038/tp.2012.61 · Cited by 480 · see citing works
Association of Nitrotyrosine Levels With Cardiovascular Disease and Modulation by Statin Therapy
Shishehbor, M. H., Aviles, R. J., Brennan, M.-L., Fu, X., Goormastic, M., Pearce, G. L., Gokce, N., Keaney, J. F., Penn, M. S., Sprecher, D. L., et al. (2003) · JAMA · doi:10.1001/jama.289.13.1675 · Cited by 464 · see citing works
Nitrite: A physiological store of nitric oxide and modulator of mitochondrial function
Shiva, S. (2013) · Redox Biology · doi:10.1016/j.redox.2012.11.005 · Cited by 207 · cited 1× · see citing works
“In biological fluids or buffers, NO systems are almost completely oxidized to nitrite (NO 2 ), a biologically inert metabolite of NO oxidation [19].”
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Research works printed but not linked: 13
- D. Shah, N. Mahajan, S. Sah, S.K. Nath, B. Paudyal, Oxidative stress and its biomarkers in systemic lupus erythematosus, J. Biomed. Sci. 21 (2014), 23e23.
- R. Radi, Nitric oxide, oxidants, and protein tyrosine nitration, Proc. Natl. Acad. Sci. U. S. A. 101 (2004) 4003e4008.
- N. Sucu, A. Unlü, L. Tamer, B. Aytacoglu, B. Ercan, M. Dikmengil, U. Atik, 3- Nitrotyrosine in Atherosclerotic Blood Vessels, Clinical Chemistry and Labo- ratory Medicine: CCLM/FESCC, vol. 41, 2003, pp. 23e25.
- R.K. Upmacis, Atherosclerosis: A Link between Lipid Intake and Protein Tyrosine Nitration, Lipid Insights, vol. 2008, 2008, 75e75.
- P.G. Winyard, B. Ryan, P. Eggleton, A. Nissim, E. Taylor, M.L. Lo Faro, T. Burkholz, K.E. Szab o-Taylor, B. Fox, N. Viner, R.C. Haigh, N. Benjamin, A.M. Jones, M. Whiteman, Measurement and meaning of markers of reactive species of oxygen, nitrogen and sulfur in healthy human subjects and patients with inflammatory joint disease, Biochem. Soc. Trans. 39 (2011) 1226e1232.
- D.M.F. Mendonça, S.C.S. Martins, R. Higashi, M.N. Muscara, V.M. Neto, L. Chimelli, A.M.B. Martinez, Neurofilament Heavy Subunit in Cerebrospinal Fluid: A Biomarker of Amyotrophic Lateral Sclerosis?, Amyotrophic Lateral Sclerosis, vol. 12, Official Publication Of The World Federation Of Neurology Research Group On Motor Neuron Diseases, 2011, pp. 144e147.
- Mahin D. Maines (Editor-In-Chief), [Et Al.], Chapter 17, (2012). 17.15-17.15.
- A.M. Fitzpatrick, L.A.S. Brown, F. Holguin, W.G. Teague, Levels of nitric oxide oxidation products are increased in the epithelial lining fluid of children with persistent asthma, J. Allergy Clin. Immunol. 124 (2009), 990e996.e991-999.
- R.L. Wixom, C.W. Gehrke, Chromatography: A Science of Discovery, Wiley, 2011.
- M. Naushad, M.R. Khan, Ultra Performance Liquid Chromatography Mass Spectrometry: Evaluation and Applications in Food Analysis, Taylor & Francis, 2014.
- V.R. Meyer, Practical High-performance Liquid Chromatography, Wiley, 2013.
- C. Herce-Pagliai, S. Kotecha, D.E. Shuker, Analytical methods for 3- nitrotyrosine as a marker of exposure to reactive nitrogen species: a review, Nitric Oxide Biol. Chem. Off. J. Nitric Oxide Soc. 2 (1998) 324e336.
- M.W. Dong, Modern HPLC for Practicing Scientists, Wiley, 2006.
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Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases
Kehm, R., Baldensperger, T., Raupbach, J., & Höhn, A. (2021) · Redox Biology · doi:10.1016/j.redox.2021.101901 · Cited by 348 · see citing works
“Protein carbonyls, 3-NT and methionine sulfoxide are biomarkers of protein oxidation which are commonly analyzed via immunochemical methods [75,82,92].”
Background
- Authors
- Teixeira, D., Fernandes, R., Prudêncio, C., & Vieira, M.
- Affiliations
- Centro de Investigação em Saúde e Ambiente (PT)
- Universidade do Porto (PT)
- Corresponding author
- Vieira, Mónica
- Venue
- Biochimie
- Publisher
- Elsevier BV
- Published
- 2016-06
- Volume
- 125
- Pages
- 1-11
- ISSN
- 0037-9042 (ISSN-L)
- 0300-9084
- OpenAlex
- W2278824265
- Cited by
- 90
- References
- 4
- Citations by year
- 2026: 8 · 2025: 4 · 2024: 7 · 2023: 9 · 2022: 14 · 2021: 10 · 2020: 14 · 2019: 13 · 2018: 4 · 2017: 6 · 2016: 1
- FWCI
- 0.72
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Biochimie (publishedVersion)
- PubMed (publishedVersion)
- Portuguese National Funding Agency for Science, Research and Technology (RCAAP Project by FCT) (submittedVersion, other-oa)
- Funders
- Integration into Scientific Research Grant
- Centro de Investigação em Saúde e Ambiente (CISA) (B Int-ICD/IPP-BST/CISA/01/2014)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Keywords
- nitrotyrosine, nitrosative stress, protein nitration, immunological detection, HPLC detection, LC-MS/MS, GC-MS/MS, sensitivity and specificity
- MeSH terms
- Animals, Humans, Protein Processing, Post-Translational, Tyrosine
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