Gel-based immunotest for simultaneous detection of 2,4,6-trichlorophenol and ochratoxin A in red wine

• Published in: Analytica chimica acta Vol. 672; no. 1; pp. 3 - 8
• Main Authors: Beloglazova, N.V., Goryacheva, I.Yu, Rusanova, T.Yu, Yurasov, N.A., Galve, R., Marco, M.-P., De Saeger, S.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05.07.2010
• Subjects: 2,4,6,-Trichlorophenol; Antibodies; Antibodies - immunology; Chlorophenols - analysis; Chlorophenols - immunology; Chromatography; Cleaning; Color; Colour; Food control; Gels - chemistry; Immunoassay; Mathematical analysis; Ochratoxin A; Ochratoxins - analysis; Ochratoxins - immunology; Rapid test; Red wine; TCP (protocol); Wine - analysis; Wines; Immunoassay; Rapid test; 2,4,6,-Trichlorophenol; Red wine; Ochratoxin A; Food control
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2010.05.024

A new rapid method which allows simultaneous one step detection of two analytes of different nature (2,4,6,-trichlorophenol (TCP) and ochratoxin A (OTA)) in red wine was developed. It was based on a column test with three separate immunolayers: two test layers and one control layer. Each layer consisted of sepharose gel with immobilized anti-OTA (OTA test layer), anti-TCP (TCP test layer) or anti-HRP (control layer) antibodies. Analytes bind to the antibodies in the corresponding test layer while sample flows through the column. Then a mixture of OTA-HRP and TCP-HRP in appropriate dilutions was used, followed by the application of chromogenic substrate. Colour development of the test layer occurred when the corresponding analyte was absent in the sample. HRP-conjugates bound to anti-HRP antibody in the control layer independently of presence or absence of analytes and a blue colour developed in the control layer. Cut-off values for both analytes were 2 μg L −1. The described method was applied to the simultaneous detection of TCP and OTA in wine samples. To screen the analytes in red wine samples, clean-up columns were used for sample pre-treatment in combination with the test column. Results were confirmed by chromatographic methods.