title: A direct competitive enzyme-linked immunosorbent assay by antibody coated for diethyl phthalate analysis.
abstract: A direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed for detection of diethyl phthalate (DEP). Protein-hapten conjugate was synthesized to produce polyclonal antibodies against DEP. Experimental parameters were optimized, including immunoreaction conditions, the dilution ratio of horseradish peroxidase (HRP)-antigen conjugate, time of the antibody coated, effect of pH, and ionic strength. The limit of detection was 0.096ng/ml, and the linear range was 0.1-3500ng/ml with a regression coefficient (R(2)) of 0.9957. Recoveries were between 96.4 and 106.2%. The cross-reactivities of the anti-DEP antibody to six structurally related phthalate esters were less than 9%. The method was successfully applied to the determination of DEP in tap water, river water (Yangtze River), and leachate from plastic drinking bottles. This immunoassay was highly specific, sensitive, rapid, simple, and suitable for DEP monitoring. The results obtained were compared with those obtained using the high-performance liquid chromatography method.
authors.affiliation: Anhui Normal University, Anhui Key Laboratory of Chemo-Biosensing, Wuhu, China. zhangmc@mail.ahnu.edu.cn
authors.forename: Mingcui
authors.initials: M
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Cong
authors.forename: Yali
authors.initials: Y
authors.lastname: Sheng
authors.forename: Bolin
authors.initials: B
authors.lastname: Liu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D002138:Calibration; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D010795:Phthalic Acids / Q000032:analysis* / Q000276:immunology; D013997:Time Factors; D014867:Water / Q000737:chemistry
chemical_list: D000906:Antibodies; D010795:Phthalic Acids; D014867:Water; C007379:diethyl phthalate