Abstract
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.
Highlights
Phthalate esters are in widespread use as plasticizers in the production of various consumable or household products
Many studies have confirmed that dibutyl phthalate (DBP) and other phthalate esters could migrate from food packaging materials into the food itself [2,3,4]
DBP was recently reportedly found in liquor [5,6], juice [7], vegetable oil [8], milk [9] and other foods [10]
Summary
Phthalate esters are in widespread use as plasticizers in the production of various consumable or household products. Worldwide production of phthalate esters and their frequent application in a wide range of products in daily use has resulted in their presence in all parts of the environment and, in food. The first immunoassay for phthalate ester came from Ius’s research in 1993 [21]. He developed a time-resolved fluoroimmunoassay to detect dimethyl phthalate in water. Zhang, MC has published immunoassays for various phthalates including diethyl phthalate (DEP) [22], dimethyl phthalate (DMP) [23,24], dipropyl phthalate (DPP) [25], dibutyl phthalate (DBP) [26], and dicyclohexyl phthalate (DCHP) [27,28,29]. A specific monoclonal antibody against DBP was prepared and an enzyme-linked immunosorbent assay (ELISA) was established to detect the presence of DBP in liquor. Real liquor samples were analyzed with the proposed ELISA, which showed good correspondence with liquid chromatography-mass spectrum results
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