Dulce Teixeira1, Rúben Fernandes2, Cristina Prudêncio2, Mónica Vieira3. 1. Ciências Químicas e das Biomoléculas, Centro de Investigação em Saúde e Ambiente, Escola Superior de Tecnologia da Saúde do Porto, Instituto Politécnico do Porto, Portugal. 2. Ciências Químicas e das Biomoléculas, Centro de Investigação em Saúde e Ambiente, Escola Superior de Tecnologia da Saúde do Porto, Instituto Politécnico do Porto, Portugal; I3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal. 3. Ciências Químicas e das Biomoléculas, Centro de Investigação em Saúde e Ambiente, Escola Superior de Tecnologia da Saúde do Porto, Instituto Politécnico do Porto, Portugal; I3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal. Electronic address: mav@estsp.ipp.pt.
Abstract
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.
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.
Authors: Zachary N Robinett; Girish Bathla; Angela Wu; James Jason Clark; Zita A Sibenaller; Thomas Wilson; Patricia Kirby; Bryan G Allen; Marlan R Hansen Journal: Otol Neurotol Date: 2018-10 Impact factor: 2.311
Authors: Juan García-Revilla; Isabel M Alonso-Bellido; Miguel A Burguillos; Antonio J Herrera; Ana M Espinosa-Oliva; Rocío Ruiz; Luis Cruz-Hernández; Irene García-Domínguez; María A Roca-Ceballos; Marti Santiago; José A Rodríguez-Gómez; Manuel Sarmiento Soto; José L Venero; Rocío M de Pablos Journal: J Clin Med Date: 2019-10-17 Impact factor: 4.241