Subject(s)
Diploidy , Glucosephosphate Dehydrogenase/metabolism , Hypoxanthine Phosphoribosyltransferase/metabolism , Phosphoglycerate Kinase/metabolism , Polyploidy , Sex Chromosomes/physiology , X Chromosome/physiology , Adenine Phosphoribosyltransferase/metabolism , Animals , Embryo, Mammalian , Female , Fibroblasts , Phosphogluconate Dehydrogenase/metabolism , Pyruvate Kinase/metabolism , Rabbits , X Chromosome/enzymologyABSTRACT
We describe an ultramicromethod for determination of uric acid by a manual procedure. Hydrogen peroxide, formed when uric acid is oxidized in the presence of uricase, oxidatively couples two molecules of p-hydroxyphenylacetic acid in the presence of peroxidase to produce a highly fluorescent compound. The specificity of the uricase reaction is retained, and the fluorometry results in a higher sensitivity (but a lower precision) than that obtained by applying spectrophotometric methods. As little as 50 ng of uric acid can be determined, and only 10 mul of plasma is required. Values obtained for human plasma are higher than those obtained by the spectrophotometric method.
Subject(s)
Uric Acid/blood , Ascorbic Acid/blood , Evaluation Studies as Topic , Humans , Microchemistry , Spectrometry, Fluorescence/methods , Spectrophotometry/methodsSubject(s)
Blood Platelets/metabolism , Serotonin/blood , Adolescent , Adult , Cell Separation , Centrifugation, Density Gradient , Female , Humans , Male , Middle AgedABSTRACT
An automatic micromethod is described for the estimation of plasma uric acid. The hydrogen peroxide formed when uric acid is oxidised by uricase, condenses by oxidation two molecules of p-hydroxyphenil-acetic acid in the presence of peroxidase. The product formed (dicarboxymethyl-5,5' dihydroxy-2,2' biphenyl) is fluorescent. This method permits one to estimate quantitites of uric acid of the order of 1 microng and permits measurement of uric acid in 50 microll of plasma. The values obtained for human plasma are compared to those supplied by a reductrimetric technique (SMA 12-60).
