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Peroxyoxalate chemiluminescent assay for oxidase activities based on detecting enzymatically formed hydrogen peroxide
Authors:Kenichiro Nakashima  Naotaka Kuroda  Shinki Kawaguchi  Mitsuhiro Wada  Shuzo Akiyama
Abstract:A sensitive peroxyoxalate chemiluminescent (PO-CL) assay for activities of oxidases (uricase, choline oxidase, cholesterol oxidase and xanthine oxidase) which catalyse a formation of hydrogen peroxide was developed using 4,4′-oxalyl-bis(trifluoromethylsulphonyl)imino]trimethylene-bis(4-methylmorpholinium)trifluoromethanesulphonate as a chemiluminogenic reagent and 2,4,6,8-tetramorpholinopyrimido5,4-d]pyrimidine as a fluorophore. The standard curve for hydrogen peroxide was linear over the range 1 × 10?7-1 × 10?4 mol/L. Relative standard deviations for oxidase assays were 5.1–12.7% (n = 10). Detection limits were 1 × 10?3 U/mL for uricase, 5 × 10?4 U/mL for choline oxidase, 5 × 10?3 U/mL for cholesterol oxidase and 5 × 10?4 U/mL xanthine oxidase (sample to blank ratio, 3).
Keywords:Peroxyoxalate chemiluminescent assay  oxidase  4  4′  -oxalyl-bis[(trifluoromethylsulphonyl)imino]trimethylene-bis(4-methylmorpholinium)trifluoromethanesulphonate  2  4  6  8-tetramorpholinopyrimido[5  4-d]pyrimidine
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