Continuous-Flow Assay for Urinary Oxalate Using Immobilised Oxalate Oxidase

Abstract

A continuous-flow assay for measuring oxalate in urine is described. Covalently attached oxalate oxidase (EC 1.2.3.4) is used to oxidise the oxalate anion to carbon dioxide and hydrogen peroxide. The formed hydrogen peroxide is measured colorimetrically (A_58n) with an established reaction using horseradish peroxidase (EC 1.11.17), 3-methyl-2-benzothiazolinone hydrazone (MBTH) and 3-dimethylaminobenzoic acid (DMAB). Ascorbate interference is eliminated by treating the urine sample with sodium nitrite prior to assaying. The assay is accurate (mean recovery of added oxalate in spiked urine sample is 93 ± 11%), sensitive (detection limit 1·0 μmol/L), reproducible (within-batch CV 3·5%; between-batch CV 5%) and relatively rapid (15 samples/h). This assay correlates well (R = 0·99) with another established enzymatic method (using oxalate decarboxylase).