Most reported laboratory reference values for harbor seals (Phoca vitulina) are derived from captive seals, or stranded seals that have recovered from disease in marine mammal centers. This study established hematology and serum chemistry reference values for free-ranging harbor seals, using methods and that are current and readily available, and determined the effects of hemolysis on serum chemistry values of captive harbor seals. Blood samples were collected for hematologic and serum chemistry measurements from 14 clinically normal, adult male and female harbor seals and two juvenile harbor seals (approximate age 6 mo) captured in saltwater sloughs and estuaries near Moss Landing, California, USA. Values for amylase, globulin, and differential leukocyte count, not previously reported, were determined. In general, hematology and chemistry values in adults were similar to those reported for free-ranging and captive harbor seals, except for glucose, urea nitrogen, and lactate dehydrogenase (LDH) values, which were higher than those reported previously. Red blood cell counts in the two juveniles were higher than in adults and in young harbor seals studied previously. To determine the effects of hemolysis on serum chemistry values, two intensities of hemolysis were generated experimentally in blood collected from 11 harbor seals recovering from injuries or stranding at the Marine Mammal Center (Sausalito, California 94965, USA). Moderate hemolysis (++, 1 g/L hemoglobin, red-tinged) significantly increased LDH activity, whereas severe hemolysis ( , 2 g/L hemoglobin, cherry red) significantly increased total protein, albumin, calculated globulin, LDH, and total bilirubin and significantly decreased creatinine. The effects of hemolysis must be considered when chemistry results of harbor seals are interpreted.