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KMID : 0903519930360050364
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1993 Volume.36 No. 5 p.364 ~ p.369
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Reevaluation of the effect of Triton X - 100 on the assay of superoxide radical by the nitrobluetetrazolium reduction method
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Abstract
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Triton X-100 enhances to a marked extent the analytical sensitivity of the nitrobluetetrazolium (NBT) reduction method for the assay of superoxide(O₂^+) production. In the present work, it was attempted to elucidate the physicochemical nature of this Triton X-100 effect, focusing on not only the surfactant-caused stabilization of the water-insoluble formazan colloid but also the kinetic competition between the NBT-O₂^+ reaction and the autodisproportionation of concommitantly occurring in aqueous media. The measurements of formazan and H₂O₂ formed in a number of reaction systems, as prepared by vortex-mixing potassium superoxide dissolved in an aprotic solvent with aqueous solutions of NBT, revealed that Triton X-100 exerts its effect both through preventing formazan colloid from aggregation and thereby increasing the formazan absorbance and through suppressing the autodisproportionation reaction of O₂^+. It also turned out that the relative shares of the colloid stabilization effect and the kinetic effect in the contribution to the sensitivity amplification of the NBT method are dependent upon the reaction conditions, particulary the molar concentration ratio of NBT to O₂^+ in the reaction systems.
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