Journal
CHEMOSPHERE
Volume 92, Issue 1, Pages 150-155Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.chemosphere.2013.02.062
Keywords
MEHP; Monoclonal antibody; Direct competitive ELISA; Detection
Categories
Funding
- National Nature Science Foundation of China [31071539]
- Science and Technology Project of Quality Supervision Inspection and Quarantine of Jilin Province [2011ZJK12]
- Agricultural Product Safety of Jilin Province under International Bank for Reconstruction and Development Loans [2011Y36]
- International Science and Technology Cooperation Program of China [2011DFA32230]
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Di-(2-ethylhexyl) phthalate (DEHP) is the most commonly used plasticizer for flexible polyvinyl chloride (PVC), which is also known as one of the environmental endocrine disruptors with the reproductive, developmental and embryonic toxicity after entering human body. Mono-2-ethylhexyl phthalate (MEHP) is one of the most complicate metabolites from DEHP in vivo and responsible for many toxic effects of DEHP. In order to evaluate human exposure to DEHP, a direct competitive enzyme-linked immunosorbent (dcELISA) based on monoclonal antibody (mAb) was developed to detect MEHP. A hybridoma cell line 4B9 secreting mAb against MEHP was prepared, and the horseradish peroxidase (HRP) labeled antigen as a probe in the dcELISA was made. After optimization of ELISA reaction conditions, the standard curve with a linear range from 0.56 to 1000 ng mL(-1) and a detection limit of 0.39 ng mL(-1) was established. The cross-reactivities of anti-MEHP mAb to other ten phthalate esters were less than 5% except for mono-methylphthalate (MME). The average recoveries of MEHP from distilled water and negative human urine were both between 87.4% and 94.72% with coefficient of variation (CV) less than 5%. Here, the ELISA method on detecting MEHP was successfully established and applied to real urine sample analyses and the results were confirmed by HPLC. Furthermore, it was indicated that the immunoassay was reliable and suitable for monitoring MEHP. (C) 2013 Elsevier Ltd. All rights reserved.
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