Assay of plasma oxalate with soluble oxalate oxidase.
We use oxalate oxidase from barley seedlings for the colorimetric determination of oxalate in plasma. The oxalate is converted to hydrogen peroxide, which, in the presence of peroxidase, is detected by a Trinder-like chromogenic system. Optimization of the assay, including deproteinization and elimination of interferences from reducing substrates, is described. Ascorbate additions (200 mumol/L) did not affect oxalate concentration in plasma, even after long frozen storage. Mean analytical recovery of oxalate averaged 102% +/- 6.9%, imprecision (CV) at 2.0 mumol/L was 7.2%, and the lower limit of quantification (CV = 20%) was 0.6 mumol/L. Results correlated well with those by chromatography (r = 0.999, Sy/x = 0.29 mumol/L, n = 32, range for x, y = 0-140 mumol/L). Plasma oxalate concentrations measured in 32 healthy subjects ranged from 0.6 to 2.9 mumol/L (mean 1.28, SD 0.71 mumol/L), which agrees with those measurable by using indirect radioisotopic dilution methods. Patients with primary hyperoxaluria and chronic renal failure exhibited markedly greater plasma concentrations of oxalate.
Renal Stone Laboratory, Mauriziano Umberto I Hospital, Turin, Italy.
M
M
Petrarulo
E
E
Cerelli
M
M
Marangella
D
D
Cosseddu
C
C
Vitale
F
F
Linari
D000293:Adolescent; D000327:Adsorption; D000328:Adult; D001205:Ascorbic Acid / Q000494:pharmacology; D001557:Benzenesulfonates; D001798:Blood Proteins; D002606:Charcoal; D002648:Child; D002863:Chromogenic Compounds; D003124:Colorimetry / Q000379:methods* / Q000706:statistics & numerical data; D004355:Drug Stability; D005260:Female; D005615:Freezing; D001467:Hordeum / Q000201:enzymology; D006801:Humans; D006959:Hyperoxaluria / Q000097:blood; D007676:Kidney Failure, Chronic / Q000097:blood; D008297:Male; D010070:Oxalates / Q000097:blood*; D019815:Oxalic Acid; D010088:Oxidoreductases; D011786:Quality Control; D012016:Reference Values; D012459:Salicylates; D012680:Sensitivity and Specificity
D001557:Benzenesulfonates; D001798:Blood Proteins; D002863:Chromogenic Compounds; D010070:Oxalates; D012459:Salicylates; D002606:Charcoal; D019815:Oxalic Acid; D010088:Oxidoreductases; C020598:oxalate oxidase; C003366:sulfosalicylic acid; D001205:Ascorbic Acid