Development of an Enzyme-Linked Immunosorbent Assay for Dibutyl Phthalate in Liquor

A monoclonal antibody specifically recognizing dibutyl phthalate (DBP) was prepared based on a hapten (di-n-butyl-4-aminophthalate). After optimizing various parameters such as concentrations of antibody, coating antigen and composition of the assay buffer, an inhibition curve was plotted with the 50% inhibition concentration value (IC50) 33.6 +/- 2.5 ng/mL. A low level of cross-reactivity (<5%) was found for other phthalate esters. Recovery tests were conducted using liquor simulant (a mixture of water and ethanol) at two fortification levels (100 ng/mL and 300 ng/mL). The recovery rates ranged from 84.7% to 94.5% with a coefficient of variation between 7.1% and 12.8%. Nine liquor samples of different alcoholic strengths were detected using the proposed measure and confirmatory analysis was performed using liquid chromatography-mass spectroscopy (LC-MS). The detection results showed good consistency between the two measures and all the data above indicated that the proposed ELISA could be applied in DBP screening.