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Development of a solid-phase extraction—enzyme-linked immunosorbent assay method for the determination of estrone in water
Authors:Zhilan Li  N Alice Lee  Ivan R Kennedy
Affiliation:a Faculty of Agriculture, Food and Natural Resources, The University of Sydney, Sydney, NSW 2006, Australia
b Department of Pharmacology, The University of Sydney, Sydney, NSW 2006, Australia
c Faculty of Food Science and Bioengineering, Tianjin University of Science and Technology, Tianjin 300222, PR China
Abstract:Estrone has been identified as a potential endocrine-disrupting chemical (EDC). To facilitate its analysis, a highly selective and sensitive enzyme-linked immunosorbent assay (ELISA) method with a simple solid-phase extraction (SPE) for analysis of estrone in aquatic environments has been developed. The specific polyclonal antibody was produced against a conjugate of estrone-3-hemisuccinate and keyhole limpet hemocyanin (KLH). The obtained ELISA showed specific recognition of estrone, without cross-reactions for three other major estrogenic compounds (17β-estradiol, estriol, and 17α-ethynylestradiol) commonly found in water. The ELISA had a limit of detection of 0.14 μg/l estrone in water. Combining a SPE method to extract and pre-concentrate estrone from water samples and ELISA to specifically quantify estrone content, the SPE-ELISA can detect estrone down to 1.25 ng/l level in water. Good recovery with spiked river water was obtained with this SPE-ELISA method. The developed SPE-ELISA system was applied to analyze the real influent and effluent samples of sewage treatment plant in Penrith (Australia) and the results correlated well with those obtained using GC and HPLC methods. The developed SPE-ELISA method is capable of being applied for the specific detection and routine monitoring of estrone in environmental water samples.
Keywords:Estrone   ELISA   Solid-phase extraction   Water
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