PARENT SESSION

1A - Environmental Analytical Methods
Hall 8
8:30 AM - 12:30 PM, Monday, 28 April 2003
Chair: Schäffer, A.¹, ¹
Co-chair: Bester, K.², ²

(MO8/7) Development of ELISAs for Quantitative Analysis for Environmental Hormones and Surfactants.

Goda, Yasuhiro¹, Kobayashi, Ayako¹, Hirobe, Masato¹, Fujimoto, Shigeru¹, Ike, Michihiko², Fujita, Masanori², Okayasu, Yuji³, Komori, Koya³, Tanaka, Hiroaki³, ¹ Takeda Chemical Industries Ltd., Osaka, Osaka, Japan² Osaka university, Suita, Osaka, Japan³ Independent Administrative Institution Public Works Research Institute, Tsukuba, Ibaragi, Japan

ABSTRACT- For quantification of ppb or ppt concentration of environmental pollutants, instrument analysis like GC-MS, GC-MS/MS, LC-MS, LC-MS/MS are usually employed. While these methods can be reliable, 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 highly sensitive, simple to perform, rapid and cost-effective method such as enzyme-linked immunosorbent assay (ELISA). Eight kinds of ELISAs were developed for environmental hormones, i.e. BPA (bisphenol A) (regular and super sensitive: ss), AP (the total amount of alklyphenol and alkylphenol ethoxylate), APE (alkylphenol ethoxylate) and estrogens, i.e. E2 (17 beta-estradiol), E1 (estrone), ES (the total amount of E1, E2 and E3 (estriol)), EE2 (17alfa-ethynylestradiol). And 2 kinds of ELISAs for surfactants, i.e. AE (alkyl ethoxylate) and LAS (linear alkylbenzene sulfonates) were also developed. The lowest limit of quantification for BPA, ss-BPA, AP, APE, E2, E1, ES, EE2, AE, LAS were 5, 0.05, 5, 20, 0.05, 0.05, 0.1, 0.05, 20 and 20 ppb, respectively. Specificity of each ELISA was confirmed with the cross-reaction test. A simplified pre-treatment protocol was also developed for sample matrices: solid phase extraction for environmental water and solvent extraction for serum. In the measurement of cleaned and concentrated samples, the values of ELISA showed a high correlation with those of conventional instrument analysis and the bias between these two methods were reasonably suppressed. The ELISAs described here are now available from Takeda (http://www.takeda.co.jp/l-e/elisa/index-e.html) and ELISAs for DEHP (diethyl phthalate), DBP (dibutyl phthalate), alkylphenol (specific to alkylphenol) and CP (chlorophenol) are under development. This study was partly supported by a grant-in-aid from the Research Institute of Innovative Technology for Earth (RITE) in Japan.

Key words: environmental hormones, ELISA, surfactants, analysis