Urinary iodide determined by paired-ion reversed-phase HPLC with electrochemical detection

Abstract

We propose an automated method for the routine analysis of urinary iodide, using paired-ion reversed-phase HPLC with electrochemical detection and a silver working electrode. Assay conditions include a flow rate of 1.0 mL/min and an operating potential of 0.10 V. The retention time for iodide is 5.4 min. Sample preparation can be semiautomated by use of a reduced-pressure manifold. The detection threshold (signal-to-noise ratio of 3) was 2 pmol, corresponding to 0.04 mumol/L. The within-run precision (CV) for a pooled urine sample was 3.9% at 452 nmol/L iodide. The average recovery of added iodine was 94%. For comparison with a colorimetric method, we measured 177 random (untimed) urine samples by both HPLC (y) and a Technicon AutoAnalyzer acid digestion method (x). After removal of organically bound iodine, the results for unbound urinary iodide determined by the two methods were nearly identical (r = 0.99; y = -0.03 + 1.00x; Sy/x = 0.12 mumol/L). Comparison of total urinary iodine measured by the Technicon AutoAnalyzer with unbound urinary iodide determined by HPLC also showed a high correlation (r = 0.96; y = -0.03 + 0.78x; Sy/x = 0.23 mumol/L), because iodine is excreted in urine mainly as iodide. We conclude that iodine in urine can be accurately determined by the more convenient HPLC assay.