Chemiluminescence of Cypridina Luciferin Analogs. Part 3. MCLA Chemiluminescence with Singlet Oxygen Generated by the Retro‐Diels‐Alder Reaction of a Naphthalene Endoperoxide

The chemiluminescence of the Cypridina luciferin analog, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroirnidazo[1,2-a]pyrazin-3-one (MCLA), with O2 (1Δg) generated by the retro-Diels-Alder reaction of 3-(4′-methyI-l'-naphthyl)-propionic acid endoperoxide was studied in an aqueous solution with pH 7.12 at 37°C. The retro-Diels-Alder reaction occurs with a first-order rate constant of (4.16 ± 0.13) × 10−4/s to quantitatively yield O2 (1Δg) and 3-(4′-methyl-l'-naphthyl (-propionic acid. MCLA consumed equimolar amounts of O2 (1Δg) with a second-order rate constant (6.96 ± 0.27) × 108/M/s to emit light in an aqueous solution with pH 7.12 at 37°C. The chemiluminescence spectrum was identified as the fluorescence spectrum of 2-acetylamino-5-(p-methoxyphenyl)pyrazine (OMCLA), a major chemiluminescence reaction. Chemiluminescence spectra and product yields for MCLA reactions with O21Δg, with O2 (3Σ−g) and with superoxide anion radicals are identical, suggesting that all of these reactions occur via a common MCLA-2-hydrope-roxide intermediate formed by a combination of MCLA radicals and superoxide anion radicals. We have established practical use of NEPO as an O2 (1Δg) source and MCLA as a biological probe for detecting O2 (1Δg).