In order to evaluate the ecological hazard of 4-tert-butylphenol (4-TBP), species abundances and physicochemical properties were monitored for 63 days in a microcosm system containing planktonic algae, rotifers, shrimps and Daphnia. The 63 days-NOEC (no observed effect concentration) of 4-TBP for this system was calculated. At the same time, species sensitivity distribution curves were constructed based on the toxicity data of EC₁₀, EC₅₀ and NOEC derived from laboratory single-species toxicity tests, which further gives no obvious hazard concentrations for 95% species in the ecosystem (HC₅). In both the microcosm and the single-species tests, the shrimp (Neocaridina sp.) was the most sensitive species to 4-TBP exposure, while most algae species were relatively insensitive compared to the zooplankton. For the microcosm exposed to 4-TBP, significant changes occurred to the community structure instead of the community function, resulting in a NOEC_microcosm of 265.38 μg L^–1 which approached the EC₁₀-based HC₅ (0.376 mg L^–1). Nevertheless, EC₅₀-based HC₅ was higher than the NOEC_microcosm, and the NOEC-based HC₅ was lower. The data showed that the EC₁₀-based HC₅ was protective at the similar level with the result of microcosm for 4-TBP, providing supportive data for chemical risk assessment.