Using C.I. Reactive Yellow 3 as the target compound, the effect of the combined use of ozonation and post-biodegradation on the decrease in dissolved organic carbon (DOC) concentration was investigated, and the synergistic effect (the difference in the amounts of DOC removed by the biological process between solutions with and without ozonation) was estimated. A decrease in DOC concentration was observed during ozonation and ΔO3/ΔDOC was decreased from 16.0 to 5.2 with increasing ozonation time. Moreover, an enhancement of biodegradability was shown. A further decrease in DOC concentration was observed during the biodegradation after ozonation. The total amount of DOC removed by the combined method was increased from 73.6 mg at 30 min to 159.9 mg at 4 h. The synergistic effect was in the range of 22.7 to 39.2 mg. BOD5 was a better indicator of the synergistic effect than BOD5/DOC.