F. William Sunderman1, Shozo Nomoto1
1Department of Laboratory Medicine, University of Connecticut School of Medicine, Hartford, Conn. 06112
Tóm tắt
Abstract
Optimum reaction conditions were evaluated for assay of serum ceruloplasmin by measurement of its p-phenylenediamine oxidase activity. The pH optima of p-phenylenediamine oxidase activities of human and rat ceruloplasmins were 5.45 and 5.2, respectively. The p-phenylenediamine oxidase activity of rat ceruloplasmin was markedly inhibited by phosphate (0.1 mol/liter), that of human ceruloplasmin only slightly. These reaction conditions were judged to be optimum for the ceruloplasmin assay: (a) substrate: p-phenylenediamine dihydrochloride, 8.9 mmol/liter; (b) buffer: acetate, 0.1 mol/liter; (c) chloride concentration: 21 mmol/liter; (d) serum dilution: 31-fold; (e) incubation: 37°C for 30 min; (f) spectrophotometry: 530 nm (vs. a "serum blank" containing NaN3). Using these reaction conditions, we devised an improved technique for measuring serum ceruloplasmin. The coefficients of variation of replicate analyses by this technique were 1.25% (for "within-run" precision) and 2.8% (for "day-to-day" precision). The mean concentration of ceruloplasmin in sera from 29 healthy men was 0.315 g/liter (SD = ± 0.049, range = 0.233 to 0.402).
