REESTIMATION OF CYPRIDINA LUCIFERIN ANALOGS (MCLA) AS A CHEMILUMINESCENCE PROBE TO DETECT ACTIVE OXYGEN SPECIES-CAUTIONARY NOTE FOR USE OF MCLA-

KAMBAYASHI Yasuhiro, OGINO Keiki

doi:10.2131/jts.28.139

Cypridina luciferin analogs have been widely used as a specific chemiluminescence probe for the detection of superoxide anion (O₂^•−) and singlet oxygen (¹O₂). However, light emission during the reaction of Cypridina luciferin analogs and other active oxygen species (AOS) has not been reported in detail. Therefore, we re-evaluated 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazine-3-one (MCLA), one of the Cypridina luciferin analogs, as a chemiluminescence probe to detect various AOS. MCLA-dependent chemiluminescence was observed when MCLA was incubated with the following systems; 1) hypoxanthine plus xanthine oxidase (O₂^•−), 2) thermolysis of endoperoxide (¹O₂), 3) hydrogen peroxide plus ferrous ion (hydroxyl radical), 4) ferrous ion, 5) thermolysis of azo compound (alkyl peroxyl radical) and 6) hydrogen peroxide. Superoxide dismutase inhibited MCLA-dependent chemiluminescence observed during ferrous ion-induced decomposition of hydrogen peroxide. Alkyl peroxyl radical reacted with MCLA, but light was not emitted when the concentration of MCLA was high. These results suggest that radicals, except O₂^•−, appeared not to be direct inducers of MCLA-dependent light emission. In summary, MCLA-dependent chemiluminescence was induced by various AOS in addition to O₂^•− and ¹O₂, but active species must be O₂ ^•− and ¹O₂ in many cases. These points should be appreciated when Cypridina luciferin analogs, such as MCLA, are used for the detection of AOS.<br>

REESTIMATION OF CYPRIDINA LUCIFERIN ANALOGS (MCLA) AS A CHEMILUMINESCENCE PROBE TO DETECT ACTIVE OXYGEN SPECIES-CAUTIONARY NOTE FOR USE OF MCLA-

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