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Polarization and Depolarization Current (PDC) technique is an effective tool to assess the condition of oil-paper insulation system in power transformers. So far the PDC behaviors of mineral oil-paper insulation have been widely investigated. However, with the increasing number of transformer choosing natural ester as its insulation oil, it is important to investigate the PDC characteristics of natural ester-paper insulation to see whether the PDC technique can also be used to assess the condition of new insulation system using natural ester in transformers accurately. In this research, natural esterpaper insulation sample and mineral oil-paper insulation sample were subjected to thermally accelerated ageing experiment at 110°C for 120 days. The PDC characteristics of natural ester-paper insulation sample and mineral oil-paper insulation sample were compared over the ageing process. A new method for assessing the ageing condition of the oil-paper insulation in terms of the depolarization charge quantity was proposed. Results show that the polarization/depolarization current of natural ester-paper insulation sample is higher than that of mineral oil-paper insulation sample with the same ageing intervals. The depolarization charge quantity of both kinds of oil-paper insulation sample is very sensitive to their ageing conditions. There is an exponential relation between the stable depolarization charge quantity of both kinds of oil-paper insulation sample and the degree of polymerization (DP) of paper. The depolarization charge quantity can be used to predict the ageing condition of oil-paper insulation providing the measurement temperature is kept the same.