On-line dialysis and quantitative high-performance liquid chromatography analysis of iodixanol in human, rat and monkey plasma

A fully automated HPLC method for analysing the non-ionic X-ray contrast agent iodixanol in plasma samples, using on-line dialysis for sample preparation, was developed. Optimal conditions were obtained with a static dialysis donor solution of 110 μl and 4 ml of recipient solution (dialysate) pulsed onto a trace enrichment column, giving maximum 55% dialysis efficiency in less than the chromatographic run time of 20 min. Hence, one sample could be dialysed during the analysis of the previous. The maximum load of iodixanol on the trace enrichment column was 3.75 mg. Validation showed that the method was selective for iodixanol, sensitive down to 84 pmol/ml and had a high precision over a linear range up to 320 nmol/ml. Although developed for iodixanol, the method can easily be modified and applied to other substances with similar properties, i.e., substances having low protein binding and high water solubility, but strong enough stationary phase affinity to be retained by an appropriate trace enrichment column.