Peroxyoxalate chemiluminescent assay for oxidase activities based on detecting enzymatically formed hydrogen peroxide.
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[(trifluoromethyl-sulphonyl)imino]trimethylene-bis(4- methylmorpholinium)trifluoromethanesulphonate as a chemiluminogenic reagent and 2,4,6,8-tetramorpholinopyrimido[5,4- d]pyrimidine as fluorophore. The standard curve for hydrogen peroxide was linear over the range 1 x 10(-7)-1 x 10(-4) mol/L. Relative standard deviations for oxidase assays were 5.1-12.7% (n = 10). Detection limits were 1 x 10(-3) U/mL for uricase, 5 x 10(-4) U/mL for choline oxidase, 5 x 10(-3) U/mL for cholesterol oxidase and 5 x 10(-4) U/mL xanthine oxidase (sample to blank ratio, 3).
Nakashima
K
K
School of Pharmaceutical Sciences, Nagasaki University, Japan.
Kuroda
N
N
Kawaguchi
S
S
Wada
M
M
Akiyama
S
S
D000429:Alcohol Oxidoreductases / Q000032:analysis / Q000378:metabolism; D002789:Cholesterol Oxidase / Q000032:analysis / Q000378:metabolism; D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007700:Kinetics; D008163:Luminescent Measurements; D010070:Oxalates; D010088:Oxidoreductases / Q000032:analysis / Q000378:metabolism*; D014503:Urate Oxidase / Q000032:analysis / Q000378:metabolism; D014969:Xanthine Oxidase / Q000032:analysis / Q000378:metabolism
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D010070:Oxalates; C044622:peroxyoxalate; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D002789:Cholesterol Oxidase; D014969:Xanthine Oxidase; D014503:Urate Oxidase