Analytical method for determination of nitric oxide in zebrafish larvae: Toxicological and pharmacological applications

作者：Leite Carlos Eduardo^*; Teixeira Ariane da Cruz; Cruz Fernanda Fernandes; Concatto Simone Cervieri; Amaral Jefferson Henrich; Bonan Carla Denise; Campos Maria Martha; Morrone Fernanda Bueno; Oliveira Battastini Ana Maria

来源：Analytical Biochemistry, 2012, 421(2): 534-540.

DOI：10.1016/j.ab.2011.11.038

• Zebrafish larvae
• Nitric oxide
• Validation
• High-performance liquid chromatography
• Pathological conditions

Zebrafish are currently used at various stages of the drug discovery process and can be a useful and cost-effective alternative to some mammalian models. Nitric oxide (NO) plays an important role in physiology of zebrafish. The availability of appropriate analytical techniques to quantify the NO is crucial for studying its role in physiological and pathological conditions. This work aimed at establishing a high-performance liquid chromatography method for determination of NO levels in zebrafish larvae. Attempts were also made to assess the normal levels of NO at the first days postfertilization and the possible changes under pathological conditions. The method validation was quantitatively evaluated in terms of sensitivity, specificity, precision, accuracy, linearity, and recovery. NO levels from zebrafish larvae at the first days postfertilization and larvae challenged to N(G)-nitro-L-arginine methyl ester, sodium nitroprusside, Escherichia coli lipopolysaccharide, and copper sulfate were analyzed. The samples were derivatized with 2,3-diaminonaphthalene, and fluorescence detection was used for the indirect determination of NO. The method showed a good performance for all validation parameters evaluated and was efficient to monitor changes in NO concentration under physiological and pathophysiological conditions. This method might represent a powerful tool to be applied in NO studies with zebrafish larvae.

• 出版日期2012-2-15