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PT15 Alkylphenols and Alkylphenol Ethoxylates
(PT226) ELISA for Alkylphenols (AP) and Alkylphenol Ethoxylates (APE) in environmental samples.
Kobayashi, Ayako1, Goda, Yasuhiro1, Hirobe, Masato1, Fujimoto, Shigeru1, Ike, Michihiko2, Fujita, Masanori2, Rubio, Fernando3, 1 Japan EnviroChemicals, Ltd., Osaka, Osaka, Japan2 Osaka Univ., Suita, Osaka, Japan3 Abraxis LLC, Warminster, Pennsylvania, USA
ABSTRACT- Alkylphenols (AP) and Alkylphenol Ethoxylates (APE) are normally measured by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS). Conventional analytical methods need expensive instrumentation, extensive purification and technical expertise in operation. To overcome these problems and analyze large number of samples, enzyme-linked immunosorbent assay (ELISA) is a suitable alternative for AP and APE determination. We have developed a direct AP and APE ELISA for the analysis of environmental samples. The assay utilizes a specific monoclonal antibody. To generate the antibody (mAb), hybridoma cells were produced by the fusion of mouse myeloma cells and spleen cells from mice immunized with AP and APE derivatives coupled to bovine serum albumin. The assay range is 5-500 ppb (AP) and 20-1000 ppb (APE). Sensitivity at the ppt levels can be achieved by a simple solid phase extraction. The anti AP mAb was shown to be specific to AP and APnEO and their metabolites such as short ethoxy chain APnEO and alkylphenoxy carboxylic acids. And the anti APE mAb was shown to be specific to APnEO and their metabolites such as short ethoxy chain APnEO and alkylphenoxy carboxylic acids, except for nonylphenol. Moreover, no response was observed with non-APnEO surfactants as well as the other compounds with structural similarity to APnEO. Recoveries from river water were 88-112% for a spike level of 10 ppb of NP10EO after pre-concentration with a C18 solid-phase extraction. The ELISA assay was also validated by comparison with high-performance liquid chromatography (HPLC) for the analysis of APE in river samples and the correlation coefficients obtained between those methods was 0.96. Future work and challenge is to develop a monoclonal antibody specific for AP.
Key words: Alkylphenol Ethoxylate (APE), AlkylPhenol (AP), ELISA (Enzyme-linked Immunosorbent Assay), cost-effective methods
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