Plasma catecholamine assays: calibration with spiked plasma versus aqueous solutions

Both aqueous standards and plasmas spiked with standards are used for assays of plasma constituents. In a previous study, high-performance liquid chromatographic (HPLC) assays of plasma norepinephrine, epinephrine, and dopamine performed with spiked plasma reference solutions resulted in lower thresholds and thus gave higher concentrations. To verify the hypothesis that spiked catecholamines bind to plasma proteins (like physiological catecholamines) and thus escape extraction by alumina prior to HPLC, the recovery of standards in human albumin solution was compared with similar investigations using aqueous standards. Our findings confirm that (a) albumin is responsible to a great extent for the drop in recovery and (b) calibration by extraction of aqueous standards is preferable to calibration by spiked plasma for free catecholamine assays.