The terms “serum concentration” and “plasma concentration” are often used interchangeably in therapeutic drug monitoring, but few studies have addressed the comparability of serum and plasma drug concentrations. Before implementing a change in the procedure for measuring aminoglycoside concentrations in our institution, we prospectively compared values for tobramycin and gentamicin concentrations measured in serum and plasma. For the 208 samples that were tested, plasma aminoglycoside concentrations were significantly lower than those in serum. This relationship was similar for to bramycin and gentamicin, and was unaffected by the presence of concurrent therapy with ticarcillin. This difference, while statistically significant, was not, in our estimation, of sufficient magnitude to be of clinical importance. Plasma aminoglycoside concentrations may be considered equivalent to serum concentrations and, owing to the shorter processing time involved, appear to be the preferred medium for measuring these values.