Objective: The purpose of our study was to validate iodine quantification in a phantom study with dual-source dual-energy CT (DECT) and to apply this technique to differentiate benign periablational reactive tissue from residual tumour in VX2 carcinoma in rabbits after radio-frequency ablation (RFA). Methods: We applied iodine quantification with DECT in a phantom and in VX2 carcinoma in rabbits after incomplete RFA to differentiate benign periablational reactive tissue from residual tumour and evaluated its efficacy in demonstrating response to therapeutic RFA. A series of tubes containing solutions of varying iodine concentration were scanned with DECT. The iodine concentration was calculated and compared with known true iodine concentration. Triple-phase contrast-enhanced DECT data on 24 rabbits with VX2 carcinoma were then assessed at Day 3 (n=6), 1 week (n=6), 2 weeks (n=6) and 3 weeks (n=6) after incomplete RFA independently by 2 readers. Dual-energy postprocessing was used to produce iodine-only images. Regions of interest were positioned on the iodine image over the lesion and, as a reference, over the aorta, to record iodine concentration in the lesion and in the aorta. The pathological specimens were sectioned in the same plane as DECT imaging, and the lesion iodine concentration and lesion-to-aorta iodine ratio of residual tumour and benign periablational reactive tissue were assessed. Results: There was excellent correlation between calculated and true iodine concentration (r=0.999, p<0.0001) in the phantom study. The lesion iodine concentration and lesion-to-aorta iodine ratio in residual tumour were significantly higher than in benign periablational reactive tissue in the 2-week group during the arterial phase (AP) (p<0.01) and in the 3-week group during both the AP (p<0.05) and the portal venous phase (p<0.05). There was no significant difference between them with respect to the lesion iodine concentration or lesion-to-aorta iodine ratio in the 3-day and 1-week groups. Conclusion: Iodine quantification with DECT is accurate in a phantom study and can be used to differentiate benign periablational reactive tissue from residual tumour in VX2 carcinoma in rabbits after RFA. Advances in knowledge: Iodine quantification with DECT may help in differentiating benign periablational reactive tissue from residual tumour in VX2 carcinoma in rabbits after RFA.