A multipumping flow system for in vitro screening of peroxynitrite scavengers

• Type: Article in International Scientific Journal
• Date: 2007

Authors

Marta F T Ribeiro (Author) Other; Ana C B Dias (Author) Other; Joao L M Santos (Author) FFUP; Eduarda Fernandes (Author) FFUP; Jose L F C Lima (Author) FFUP; Elias A G Zagatto (Author) Other

Journal

Title: Journal of Biomolecular Screening

Vol. 12 No. 6

Pages: 875-880

ISSN: 1087-0571

Publisher: SAGE Publications

Indexing

ISI Web of Knowledge - 8 Citations

Scopus - 8 Citations

Scientific classification

FOS: Natural sciences > Chemical sciences

Other information

Authenticus ID: P-004-7V3

DOI: 10.1177/1087057107305402

Abstract (EN): 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 6