Determination of Dimethyl Phthalate in Environment Water Samples by a Highly Sensitive Indirect Competitive ELISA

Recent controversy over the discovery of clouding agents containing the banned chemical di(2-ethylhexyl) phthalate in beverages in 2011 in Taiwan has caused public concerns. For the detection of dimethyl phthalate (DMP) in environment water samples, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed in this paper. Dimethyl 4-aminophthalate (4-DMAP) was covalently attached to bovine serum albumin as immunogen by a diazotization method. The conjugation of DMAP and ovalbumin as coating antigen was obtained in the same way. Polyclonal antibody was obtained from New Zealand white rabbits. Under the optimized conditions, DMP was detected in the concentration range of 0.02-419 ng/mL with a detection limit of 0.01 ng/mL. The proposed method has been applied to the analysis of river water, lake water, and rain water samples. Satisfactory recoveries were obtained ranging from 90.6% to 105.5%. The cross-reactivities of the anti-DMP antibody to seven structurally related phthalate esters were below 10%. The data demonstrated that the ic-ELISA method described in our study is a simple, sensitive, and specific method and showed that this assay is a reliable tool to detect DMP in water samples.