Mr. Slaughter et Pj. O'Brien, Fully-automated spectrophotometric method for measurement of antioxidant activity of catalase, CLIN BIOCH, 33(7), 2000, pp. 525-534
Objectives: Develop fully automated assay of antioxidant catalatic activity
of catalase.
Design and methods: The assay is based on standard, clinical chemistry auto
mated analyzer methods for measuring hydrogen peroxide by using the Trinder
reagent. Catalase competes with 324 U/L horseradish peroxidase (type XII)
and Trinder reagent for hydrogen peroxide produced by 46 U/L uricase action
on urate. Unit activity is defined as 50% inhibition of maximal color deve
lopment.
Results: Within-run coefficients of variation (cv) were 2% for standards an
d samples, whereas between-run cv was 3.1% for standards and 7.3% for sampl
es. Dilutional parallelism and linearity were demonstrated for 8-fold dilut
ions of samples over the range 0.1 to 1.1 U/mL. Recovery of added catalase
was complete. Samples are stable to freezing and storage for 1 week at -80
degreesC. Activities (units/mL) ranged from 0.29 to 0.41 in human and canin
e plasma, and for erythrocytes from 48 to 70 in man, 17 to 19 in dogs, and
60 to 89 in rats. Rat liver activity (units/g wet weight) was age-dependent
and ranged from 17 to 24 at 2 months, and from 19 to 37 at 6 months.
Conclusion: The first, fully automated assay for the measurement of catalat
ic activity of catalase in plasma, erythrocytes, and liver is demonstrated
for multiple species. The assay is simple, precise, relatively inexpensive,
and rapid. Copyright (C) 2000 The Canadian Society of Clinical Chemists.