A novel flow-injection chemiluminescence (CL) method for the determination of guanine was developed. The procedure is based on the CL reaction of guanine with hydrogen peroxide in borax buffer (pH 8.5) with Co2+ as a catalyst. The calibration graph is linear within the range of 3 x 10(-7)-9 x 10(-5) g/mL. A detection limit of 1 x 10(-7) g/mL, along with a relative standard deviation of 2.23% (3 x 10(-7) g/mL guanine, n = 11), were obtained. The present procedure was applied to the measurement of guanine in urine with recoveries of 97.5-107.5%. A possible CL mechanism of the reaction system is proposed.