Time--concentration studies by high-performance liquid chromatography of metabolites removed during hemodialysis.

Reversed-phase high-performance liquid chromatography was used to study the efficiency of hemodialysis. The method, previously used to study uremic hemodialysate, was also found useful for analyzing other uremic biofluids, such as serum and urine. Samples of blood (centrifuged to obtain cell-free serum) and hemodialysate were simultaneously collected from selected patients at regular intervals during the entire 6-hour hemodialysis treatment. These samples were filtered through membranes to remove particulates and proteins and were then analyzed by use of a 60-cm muBondapak-C18 column, an aqueous sodium acetate/methanol gradient, and detection at 254 nm. The identification of components in the eluate was based on retention times, ultraviolet absorption spectra, and mass spectra. From the amounts of several metabolites in the various physiological fluid pools during hemodialysis, we concluded that they mostly originate from tissue fluids. We also observed that the concentrations of most components in serum and hemodialysate fluctuated regularly with time.