Xanthine oxidase activity: simultaneous HPLC evaluation of the "D" and "O" forms.
A new HPLC method was set up for the simultaneous evaluation of the amount of uric acid and NADH produced by incubation of tissue fractions containing xanthine oxidase, from which the activity of both type "O" (oxidase) and type "D" (dehydrogenase) xanthine oxidase can be calculated. After incubation of the enzyme fraction and ethanol extraction, HPLC analysis is directly carried out. Sensitivity of the method is high enough for the evaluation of xanthine oxidase activity at the lowest reported tissue values. The reliability of the method was tested measuring the enzyme activity in rat heart and kidney extracts.
Cighetti
G
G
Department of Medical Chemistry and Biochemistry, University of Milano.
Del Puppo
M
M
Andreetta
F
F
Kienle
M G
MG
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology; D009243:NAD / Q000032:analysis / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D014527:Uric Acid / Q000032:analysis / Q000378:metabolism; D014968:Xanthine Dehydrogenase / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000378:metabolism*
D009243:NAD; D014527:Uric Acid; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase; D007658:Ketone Oxidoreductases