Analysis of Nonylphenol: Advances and Improvements in the Immunochemical Determination Using Antibodies Raised against the Technical Mixture and Hydrophilic Immunoreagents
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- 作者：M.-Carmen Esté ; vez ; Mark Kreuzer ; Francisco Sá ; nchez-Baeza ; and M.-Pilar Marco
- 刊名：Environmental Science & Technology
- 出版年：2006
- 出版时间：January 15, 2006
- 年：2006
- 卷：40
- 期：2
- 页码：559 - 568
- 全文大小：208K
- 年卷期：v.40,no.2(January 15, 2006)
- ISSN：1520-5851
文摘
The development of an enzyme-linked immunosorbentassay (ELISA) for the detection of technical nonylphenol(NP) is reported. The preparation of specific antibodies hasbeen addressed using an immunizing hapten with a four-carbon atom spacer arm placed at the ortho position thatpreserves both the hydroxyl group and the complexity ofthe branched nonyl chain mixture of the technical NP. Thesynthesis of the immunizing hapten 5-(2-hydroxy-5-nonylphenyl)-pentanoic acid has been accomplishedthrough a four-step synthetic pathway using the NPcommercial technical mixture as the starting material.Three types of competitor haptens have also been prepareddepending on the location of the spacer arm: in orthoposition to the phenol group (type A), attached to the oxygenatom (type B), and in para position, substituting thenonyl chain (type C). Drawbacks produced by thehydrophobicity of the NP or of the hapten derivativeshave been circumvented by using a highly hydrophilic carriermolecule such as a high-molecular-weight aminodextranas a coating support for antigen in an indirect ELISA format.A reproducible and sensitive indirect competitive ELISAhas been finally obtained, reaching a limit of detection of2.3 ± 0.9 
g L-1 and an IC50 value of 29 ± 5 g L-1(both N = 16). A coefficient of variation of 11% for assaysperformed on different days (N = 5; IC50 = 30 ± 3 gL-1) demonstrates the assay reproducibility. The assay alsorecognizes the nonylphenol polyethoxylates to a differentdegree depending on the length of the ethoxylate chain.Recovery values in the range between 96 and 100% havebeen obtained using spiked blind aqueous samplesalthough the sample preparation procedure used hasbeen shown to have a great influence on the methodaccuracy. A preliminary evaluation of the analytical protocolestablished has been performed using real water samples.
g L-1 and an IC50 value of 29 ± 5 g L-1(both N = 16). A coefficient of variation of 11% for assaysperformed on different days (N = 5; IC50 = 30 ± 3 gL-1) demonstrates the assay reproducibility. The assay alsorecognizes the nonylphenol polyethoxylates to a differentdegree depending on the length of the ethoxylate chain.Recovery values in the range between 96 and 100% havebeen obtained using spiked blind aqueous samplesalthough the sample preparation procedure used hasbeen shown to have a great influence on the methodaccuracy. A preliminary evaluation of the analytical protocolestablished has been performed using real water samples.