Drug Discov Ther. 2012;6(1):44-48. (DOI: 10.5582/ddt.2012.v6.1.44)

A novel flow-injection analysis system for evaluation of antioxidants by using sodium dichloroisocyanurate as a source of hypochlorite anion.

Ichiba H, Hanami K, Yagasaki K, Tanaka M, Ito H, Fukushima T


A flow injection analysis (FIA) system for evaluation of the antioxidant activity of a compound capable of scavenging a hypochlorite anion (OCl), one of the reactive oxygen species (ROS), was developed. Aminophenyl fluorescein (APF), a fluorescence indicator of ROS, was mixed manually with the test compounds and the mixed solution was injected into the FIA system. The injected solution was reacted in-line with OCl, that was produced by using sodium dichloroisocyanurate in the presence of 0.1 M CH3CO2Na in H2O. The fluorescence intensity of fluorescein generated from non-fluorescent APF was significantly attenuated in compounds that had a scavenging effect on OCl. The precision obtained by the FIA system was satisfactory (relative standard deviation < 5.0%) and a rapid assay within 0.5 min per sample was achieved. The proposed FIA system was used to demonstrate that reduced glutathione, dithiothreitol, and 3-methyl-1-phenyl-5-pyrazolone (edaravone) showed a significant scavenging effect on OCl. Therefore, the proposed FIA system can be used as a screening assay for OCl-scavenging compounds.

KEYWORDS: Screening assay, reactive oxygen species, aminophenyl fluorescein, fluorescence, reduced glutathione

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