Enzyme-linked immunosorbent assay for bisphenol A: Assay optimization and its application for surface water analysis

Abstract

A range of ELISAs in indirect and direct formats for the determination of bisphenol A (BPA) was developed. Bisphenol A carboxymethyl ether (BPA-CME), (BPA-CPE) and 4,4-bis(4-hydroxyphenyl)valeric acid were coupled with bovine serum albumin (BSA) (immunogens for the production of polyclonal antibodies), ovalbumin (OVA) and horseradish peroxidase. In general, the indirect assay was more sensitive and specific than the direct one. Using heterologous combinations of immunoreagents in the indirect assay allowed to increase assay sensitivity and specificity. The highest sensitivity was obtained for the antibodies produced against a conjugate of BPA-CPE with BSA and a conjugate of 4,4-bis(4-hydroxyphenyl)valeric acid with OVA. The detection limit of BPA in phosphate buffer was 0.03 ng mL⁻¹. The assay developed was also the most specific towards BPA. Maximum cross-reactivity values did not exceed 11% for 4-cumylphenol, 5% for bisphenol E and 2% for bisphenol S. Finally, the developed assay was used to analyze surface water samples spiked with known amount of BPA. The assay showed good recovery values (85–109%) for surface water with mineralization level lower than 500 mg L⁻¹.

Keywords:

• Bisphenol A
• endocrine disrupters
• ELISA
• surface water

Acknowledgements

The authors express their thanks to Special organic analysis group of A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, for the analysis of water samples.