Amperometric determination of oxalate in plasma and urine by liquid chromatography with immobilized oxalate oxidase

J Chromatogr B Biomed Appl. 1994 Jun 3;656(1):29-35. doi: 10.1016/0378-4347(94)80020-0.

Abstract

A detection system for plasma and urinary oxalate involving a postcolumn enzymatic reaction and electrochemical detection is described. Oxalate oxidase was immobilized on AF-Tresyl Toyopearl 650 gel. The immobilized oxalate oxidase was packed into a stainless-steel column (10 x 4 mm I.D.) and used on-line as an immobilized enzyme reactor (IMER). The hydrogen peroxide produced by enzymatic reaction was detected amperometrically at a platinum electrode maintained at +0.5 V vs. Ag/AgCl. The HPLC separation of oxalate was carried out using a Capcell Pak C8 column and an isocratic mobile phase containing 80 mM KH2PO4 and 5 mM tetra-n-butylmammonium phosphate as an ion-pair reagent. The IMER was active and stable in the mobile phase employed. The plot of peak height against concentration of oxalate was linear in the range 0.1-1.6 mumol/ml with a correlation coefficient of 0.9989. The detection limit was 10 nmol/ml at a signal-to-noise ratio of 3. The within-day and between-day coefficients of variation were 5.3 and 7.9%, respectively.

MeSH terms

  • Chromatography, High Pressure Liquid
  • Electrochemistry
  • Enzymes, Immobilized*
  • Humans
  • Hydrogen Peroxide
  • Hydrogen-Ion Concentration
  • Oxalates / analysis*
  • Oxalates / blood
  • Oxalates / urine
  • Oxidoreductases*
  • Solutions

Substances

  • Enzymes, Immobilized
  • Oxalates
  • Solutions
  • Hydrogen Peroxide
  • Oxidoreductases
  • oxalate oxidase