Enzymatic assay of dehydrogenase substrate based on the detection of superoxide anion

Superoxide generated by pyrogallol reduces highly water-soluble tetrazolium salt to produce a soluble formazan: A simple assay for measuring superoxide anion radical scavenging activities of biological and abiological samples

“…Nevertheless, the assay lacks sensitivity and is not ideally suited to microplate measurement [17]. Other methods detecting O 2 •− include electron spin resonance (ESR) spectrometry, chemiluminescence (CL) technique and fluorescence assay [18][19][20][21]. All these methods have their own limitations [1,22,23].…”

“…WST-1 has been popularly applied to various experiments in recent years [10,11,15,19]. Since the WST-1 formazan is highly water-soluble (water-solubility is greater than 100 mM) and has a high molar absorptivity (approximately 3.7 × 10 4 at 438 nm, pH 8.0) [26], the proposed method can be widely adapted to biological and abiological samples and has high sensitivity.…”