Radioisotope techniques are well known as methods for evaluating symplastic ion absorption in roots. In the present study, a new method for evaluating symplastic cadmium (Cd) absorption in plant roots was developed using the enriched isotopes 113Cd and 114Cd. Seedlings of Solanum melongena were exposed to an enriched isotope solution of 113Cd at 25°C for 30 min. The roots were excised from each seedling and were then immersed in a cold buffer solution without Cd at 2°C for 120 min to suppress the metabolic activity of the roots. Finally, the roots were treated with a cold buffer solution containing enriched stable isotope 114Cd at 2°C for 120 min, whereby the apoplastically bound 113Cd was desorbed. We tested the validity of our method for evaluating symplastic Cd in roots compared with the conventional method based on differences in the amount of Cd absorbed at 2°C and 25°C using unlabeled Cd. There was no difference in the symplastic Cd content of the roots between the two methods. These results indicate that it is possible to evaluate the symplastic Cd content in roots using the enriched isotopes 113Cd and 114Cd.