WP2 Chemical and Biological Analysis of Endocrine Disrupting Compounds
255 Portland Ballroom
1:20 PM - 4:40 PM, Wednesday
() Development of ELISAs for quantitative analysis for endocrine disrupting compounds (estrogens, industrial chemicals and surfactants).
Goda, Y1, Hirobe, M1, Kobayashi, A1, Fujimoto, S1, Ike, M2, Fujita, M2, Ohkawa, H3, Okayasu, Y4, Shiraishi, H5, Rubio, F6, 1 Japan EnviroChemicals, Ltd., Osaka, Japan2 Osaka Univ., Suita, Japan3 Fukuyama Univ., Fukuyama, Japan4 Independent Administrative Public Works Research Institute, Tsukuba, Japan5 National Institute for Environmental Studies, Tsukuba, Japan6 Abraxis LLC, Pennsylvania, USA
ABSTRACT- For quantification of estrogens, endocrine disrupting chemicals and surfactants, instrumental analysis such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are generally employed. These analytical methods are highly reliable, however, they have several potential drawbacks including expensive instrumentation, large sample volume, extensive purification and technical expertise in operation. Due to these shortcomings, the analysis of a large number of samples may be both cost and time prohibitive. Therefore there is a strong need for rapid, simple, and cost-effective methods for quantitative analysis of these contaminants such as enzyme-linked immunosorbent assay (ELISA). In this study, ten kinds of ELISA systems were developed for the quantitative analysis of estrogens [17beta-estradiol (E2), estrone (E1), estrogen (ES: E1+E2+estriol (E3)), 17alfa-ethynylestradiol (EE2)], endocrine disrupting chemicals [alkylphenol (AP), AP+alkylphenol ethoxylate (APE), and bisphenol A (BPA)] and surfactants [linear alkylbenzene sulfonates (LAS), alkyl ethoxylates (AE), and APE]. The lowest quantification limits of these ELISAs were 0.05 g/L (E2, E1, ES and EE2), 5 
g/L (AP, AP+APE), 0.05 g/L (BPA) and 20 g/L (LAS, AE and APE) when the following standards were used: E2, E1, E2, EE2, nonylphenol (NP), NP, BPA, LAS (alkyl chain length of 12), nonylphenol ethoxylate (average-ethoxy chain length of 10), AE (alkyl and ethoxy chain lengths were 12 and 7), respectively. The specificity of each ELISA was confirmed by testing several compounds, which have structural resemblance to the compounds of interest. These ELISAs were also validated by comparing them with LC-MS/MS, LC-MS and HPLC using environmental samples. Good correlations were observed between the ELISAs and instrumental analytical methods in all cases.
Key words: estrogen, ELISA, endocrine disrupting chemicals, surfactant