We evaluated an automated dry-slide enzymatic method involving creatinine iminohydrolase for measurement of creatinine in serum with the Kodak Ektachem analyzer. The means (and SD) for three commercially available quality-control sera, analyzed during eight weeks, were 9.7 (1.0), 16.6 (0.9), and 61 (2.1) mg/L. The regression equation for 105 samples measured with the Technicon SMAC (x) and Ektachem Analyzers (y) was: y = (0.89 +/- 0.007)x + (1.7 +/- 0.3) mg/L and for 170 samples measured with the Beckman Astra (x) and Ektachem analyzers (y): y = (1.00 +/- 0.005)x - (1.9 +/- 0.16) mg/L. Sixty-one samples from renal-transplant patients showed nearly the same agreement. The enzymatic method had no interference from substances that interfere with many Jaffé methods for creatinine, including acetoacetate. The drugs cephalothin and cephoxitin did not interfere, but 5-fluorocytosine interfered significantly with creatinine in the Ektachem method. Values for several ketone-positive sera were 5 to 10 mg/L higher by the Astra relative to the Ektachem. Grossly hemolyzed or lipemic samples were analyzed without difficulty. We conclude that this enzymatic method for creatinine in serum has the speed and precision necessary for routine clinical laboratory use and, except for one drug, the method appears to be specific for creatinine.