Rainbow trout: (Salmo gairdneri) anesthetized in 100 mg/liter of M.S. 222 at 12 C excreted the drug in free and acetylated forms via the urine during a 24-hr recovery period in freshwater. Of the M.S. 222 excreted, 77–96% was acetylated. Blood levels of free drug in anesthetized trout approximated 75% of the anesthetic concentration, but the amount of acetylated M.S. 222 was relatively insignificant. The blood and urine were cleared of the two fractions of M.S. 222 in 8 and 24 hr respectively. Low levels of aromatic amines of natural origin occurred in blood and urine and were subtracted from measurements of M.S. 222. Intraperitoneal injections of 10–100 mg/kg of M.S. 222 did not induce anesthesia; however, the 24-hr pattern of drug excretion was similar to that observed after anesthesia by immersion. Only 15–21% of the injected dose was found in the urine, suggesting a second route of drug elimination.