Peroxyoxalate chemiluminescent assay for oxidase activities based on detecting enzymatically formed hydrogen peroxide

✍ Scribed by Nakashima, Kenichiro ;Kuroda, Naotaka ;Kawaguchi, Shinki ;Wada, Mitsuhiro ;Akiyama, Shuzo

• Publisher: John Wiley and Sons
• Year: 1995
• Weight: 459 KB
• Volume: 10
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170100307

✦ Synopsis

A sensitive peroxyoxalate chemiluminescent (PO-CL) assay for activities o f 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)trif luoromethanesulphonate as a chemiluminogenic reagent and 2,4,6,8-tetramorpholinopyrimido[5,4-d]pyrimidine as a fluorophore. The standard curve for hydrogen peroxide was linear over the range 1 x 10-7-1 x 10-4mol/L. Relative standard deviations for oxidase assays were 5.1-12.7% (n = 10). Detection limits were 1 x U/mL for choline oxidase, 5 x U/mL xanthine oxidase (sample t o blank ratio, 3).