Discrete analysis of plasma oxalate with alkylamine glass bound sorghum oxalate oxidase and horseradish peroxidase
| Autor: | L. Goyal, Chandra Shekhar Pundir, Manisha Thakur |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Adult
Male Adolescent Immobilized enzyme Oxalate oxidase Inorganic chemistry Biophysics Poaceae Biochemistry Horseradish peroxidase Oxalate chemistry.chemical_compound Humans Phenol Child Horseradish Peroxidase Detection limit Oxalates biology Temperature Reproducibility of Results Hydrogen-Ion Concentration Middle Aged Kinetics Models Chemical chemistry Oxalate Measurement biology.protein Female Glass Oxidoreductases Peroxidase |
| Zdroj: | Journal of Biochemical and Biophysical Methods. 44:77-88 |
| ISSN: | 0165-022X |
| DOI: | 10.1016/s0165-022x(00)00071-3 |
| Popis: | We have reported a simple method of determination of plasma oxalate using a Cl(-) and NO(3)(-) insensitive oxalate oxidase purified from grain sorghum leaf and commercially available peroxidase from horseradish [Pundir et al., Ind. J. Biochem. Biophys., 35 (1998) 120-122]. The present report describes the immobilization of both the enzymes onto alkylamine glass, their kinetic properties and application for discrete analysis of plasma oxalate. In the analytic method, H(2)O(2) generated from plasma oxalate by immobilized oxalate oxidase is measured colorimetrically at 520 nm by oxidative coupling with 4-aminophenazone, and phenol catalyzed by immobilized peroxidase. The minimum detection limit of the method is 2.5 micromol/l. Analytic recovery of added oxalate in plasma was 89. 5+/-4.1% (mean+/-S.D.). The within and between day CV for plasma oxalate measurement were9.37 and11.0%, respectively. The normal range of plasma oxalate as measured by the present method was 3.6 to 5.7 micromol/l. The method is not only free from interference by plasma Cl(-) and NO(3)(-) but also provides the reuse of glass beads and thus reduces the cost of analysis for routine. |
| Databáze: | OpenAIRE |
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