Determination of oxalic acid in urine by high-performance liquid chromatography

Abstract

A new method for determination of oxalic acid in urine is described.The method encloses sample purification prior to the treatment by High Performance Liquid Chromatography (HPLC).The purification step consists in the passage of acidified urine through Sep-pak C18 cartridge (Waters), followed by the precipitation of the oxalic acid eluted with CaCl2, new dilution of the calcium oxalate precipitate, oxalic acid extraction with diethyl ether and total dryness of the sample.The losses of oxalic acid during this process are evaluated by the addition of oxalic acid (U-14C) before the precipitation step.The dried samples are redissolved in mobile phase (o-H3PO4, 0.05 M) and injected into a HPLC chromatograph, with reversed phase column (Lichrosorb RP-8, Merck).Oxalate peak is detected spectrophotometrically at 220 nm, with a retention time of 3.20 minutes.The method shows a mean recovery value of 98.25%, with an intra-run and between-run values of 5.13 and 8.06 respectively.The oxalic acid measured in urine by this method is 35.52 +/- 9.42 mg/24 h in normal subjects.