Nine kinds of hetero-chitooligosaccharides (hetero-COSs) with relatively higher molecular weights (90, 75 and 50-HMWCOS), medium molecular weights (90, 75 and 50-MMWCOS), and lower molecular weights (90, 75 and 50-LMWCOS) were prepared from partially deacetylated hetero-chitosans (90, 75 and 50% deacetylated chitosan), and their scavenging activities were investigated against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, superoxide and carbon-centered radicals using electron spin resonance (ESR) spin-trapping technique. Superoxide, hydroxyl and carbon-centered radicals were generated from hypoxanthine-xanthine oxidase reaction, hydrogen peroxide-ferrous sulfate (Fenton reaction) and azo compound 2,2-azobis-(2-amidinopropane)-hydrochloride (AAPH), respectively. The ESR results revealed that 90-MMWCOS, which is having relatively medium molecular weights prepared from 90% deacetylated chitosan, showed the highest scavenging activity on all tested radicals. In addition, the radical scavenging activity of hetero-COSs increased with a dose-dependent manner, and it was dependent on their degree of deacetylation values and molecular weights.