Increased awareness of phthalic acid esters (PAEs) toxicity has given rise to a dramatic increase in concern about the determination of these contaminations in the environment. In this paper, a sensitive, selective and rapid enzyme immunoassay of ELISA based on polyclonal antibody for detecting butyl benzyl phthalate (BBP) was developed and applied in the environmental water and soil samples. The hapten of BBP was synthesized, then applied to prepare artificial antigen and produce polyclonal antibody capable of specific recognizing BBP. From the optimal standard curve of ELISA for BBP, the values of LOD (limit of detection, IC10) and IC50 were 2.5 and 79.4 ng/mL, respectively. The ELISA showed high specificity, with the cross-reactivity toward BBP analogs < 9.6%. The satisfactory accuracy and precision were demonstrated by the recoveries of 76-116% and coefficient of variations (CVs) of 4.7-13.7%. Furthermore, BBP contamination was investigated at 3.1-25.2 ng/mL in real water samples and 4.2-76.4 ng/g in real soil samples (with the detection rate of 55% in 20 samples) by the developed ELISA, which also had shown a good correlation with that the results obtained by HPLC. All of this indicated that the developed enzyme immunoassay could be applied for sensitive and selective determination of BBP contamination in the environmental samples. Furthermore, the strategy of BBP hapten synthesis and an alternative method of BBP determination could be provided.