We discussed the relation between aquatic toxicity and interfacial activity during biodegradation with using LAS (Linear Alkylbenzene Sulphonate) and AE (Alcohol Ethoxylate). The change of death rate of Daphnia magna, surface tension, concentration of surfactant, and biodegradation by oxygen demand during biodegradation were measured. As a result, a rapid decrease in toxicity and rapid increase in surface tension were observed within the time before biodegradation based on oxygen demand started to increase. These rapid changes in toxicity and surface tension occurred due to the structural change of surfactant molecules in the primary biodegradation process, which was confirmed by HPLC (High Performance Liquid Chromatography) analysis. We also performed re-addition test to study the effects of acclimatization since it takes an important role on boidegradation, and found that the acclimatization significantly accelerated the primary biodegradation, which were indicated by increase in surface tension and decrease in aquatic toxicity. These results show that the environmental risk of surfactants should be considered not only with the biodegradation based on oxygen demand but also with the decrease of interfacial activity through the primary biodegradation process.