ESS - CQB - Ciências Químicas e das Biomoléculas
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Browsing ESS - CQB - Ciências Químicas e das Biomoléculas by Subject "3-Nitrotyrosine"
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- Development of a new HPLC-based method for 3-nitrotyrosine quantification in different biological matricesPublication . Teixeira, Dulce; Prudêncio, Cristina; Vieira, MónicaThe nitration of tyrosine residues in proteins is associated with nitrosative stress, resulting in the formation of 3-nitrotyrosine (3-NT).1 3-NT levels in biological samples have been associated with numerous physiological and pathological conditions. Hence several attempts have been made in order to develop methods that accurately quantify 3-NT in these matrices.The aim of this study was to develop a simple, rapid, low-cost and sensitive high-performance liquid chromatography (HPLC)-based 3-NT quantification method. Methods All experiments were performed on an Hitachi LaChrom Elite® HPLC system. The method was validated according to International Conference on Harmonisation (ICH) guidelines for serum samples. Additionally, other biological matrices were tested, namely whole blood, urine, B16 F-10 melanoma cell line, growth medium conditioned with the same cell line, bacterial and yeast suspensions. Results From all the protocols tested, the best results were obtained using 0.5% CH3COOH:MeOH:H2O (15:15:70) as mobile phase, with detection at wavelengths 215, 276 and 356 nm, at 25 °C, and using a flow rate of 1 mL min−1. By using this protocol, it was possible to obtain a linear calibration curve, limits of detection and quantification in the order of μg L−1, and a short analysis time (<15 min per sample). The developed protocol allowed the successful detection and quantification of 3-NT in all biological matrices tested, with detection at 356 nm. Conclusion This method, successfully developed and validated for 3-NT quantification, is simple, cheap and fast. These features render this method a suitable option for analysis of a wide range of biological matrices, being a promising useful tool for both research and diagnosis activities.
- Development of a new method for the detection and quantification of octopamine and 3-nitrotyrosinePublication . Teixeira, Dulce; Prudêncio, Cristina; Azenha, Manuel; Vieira, MónicaNeurodegenerative diseases affect around 30 million people worldwide, thus new methods that can facilitate research of such diseases are essential. Neurotransmitters, characterised as critical regulators of several neurological disorders, have been the target for the development of such methods. Besides, oxidative/ nitrosative stress has been associated with several neurodegenerative diseases and is thought to play an important role in their pathogenesis. Therefore, the development of bioanalytical methods for quantification of neurotransmitters and oxidative stress-associated molecules seems to be an interesting approach. The aim of this study was to develop a simple, rapid, low-cost and sensitive octopamine (OCT) and 3-nitrotyrosine (3-NT) quantification method for use in biological models.