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A new method was developed for the determination of lomefloxacin by using flow-injection chemiluminescence( CL) based on the reaction of Ce(IV)-sodium thiosulfate-lomefloxacin-sulfuric acid system. The CL conditions were investigated and optimized. The CL intensity was correlated linearly with concentration of lomefloxacin in the range of 1.0x10-5∼6.0x10-2 g L-1. The determination limit (3σ) is 3x10-6 g L-1. The relative standard deviation is 1.9% for 11 measurements of 2.0x10-4 g L-1 lomefloxacin. The method has been applied satisfactorily to determine lomefloxacin in drugs, serum and urine with the recovery of 97.0%∼104.5%. The CL mechanism of the system was proposed.