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Journal of Biomolecular Screening
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A Multipumping Flow System for In Vitro Screening of Peroxynitrite Scavengers

Marta F.T. Ribeiro

Departamento de Química-Física, Faculdade de Farmácia da Universidade do Porto, Porto, Portugal, joaolms{at}ff.up.pt

Ana C.B. Dias

Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, São Paulo, Brazil

Joao L.M. Santos

Departamento de Química-Física, Faculdade de Farmácia da Universidade do Porto, Porto, Portugal

Eduarda Fernandes

Departamento de Química-Física, Faculdade de Farmácia da Universidade do Porto, Porto, Portugal

José L.F.C. Lima

Departamento de Química-Física, Faculdade de Farmácia da Universidade do Porto, Porto, Portugal

Elias A.G. Zagatto

Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, São Paulo, Brazil

Peroxynitrite anion is a reactive nitrogen species formed in vivo by the rapid, controlled diffusion reaction between nitric oxide and superoxide radicals. By reacting with several biological molecules, peroxynitrite may cause important cellular and tissue deleterious effects, which have been associated with many diseases. In this work, an automated flow-based procedure for the in vitro generation of peroxynitrite and subsequent screening of the scavenging activity of selected compounds is developed. This procedure involves a multipumping flow system (MPFS) and exploits the ability of compounds such as lipoic acid, dihydrolipoic acid, cysteine, reduced glutathione, oxidized glutathione, sulindac, and sulindac sulfone to inhibit the chemiluminescent reaction of luminol with peroxynitrite under physiological simulated conditions. Peroxynitrite was generated in the MPFS by the online reaction of acidified hydrogen peroxide with nitrite, followed by a subsequent stabilization by merging with a sodium hydroxide solution to rapidly quench the developing reaction. The pulsed flow and the timed synchronized insertion of sample and reagent solutions provided by the MPFS ensure the establishment of the reaction zone only inside the flow cell, thus allowing maximum chemiluminescence emission detection. The results obtained for the assayed compounds show that, with the exception of oxidized glutathione, all are highly potent scavengers of peroxynitrite at the studied concentrations. (Journal of Biomolecular Screening 2007:875-880)

Key Words: peroxynitrite • scavenging activity • flow analysis • multipumping • chemiluminescence

References

Journal of Biomolecular Screening, Vol. 12, No. 6, 875-880 (2007)
DOI: 10.1177/1087057107305402
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This Article
Right arrow Abstract Freely available
Right arrow Free Full Text (Free PDF) Free
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
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Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
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Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Ribeiro, M. F.T.
Right arrow Articles by Zagatto, E. A.G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ribeiro, M. F.T.
Right arrow Articles by Zagatto, E. A.G.
Right arrowPubmed/NCBI databases
*Compound via MeSH
*Substance via MeSH
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?