Gel-based immunotest for simultaneous detection of 2,4,6-trichlorophenol and ochratoxin A in red wine.
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 microg 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.
Beloglazova
N V
NV
Saratov State University, Chemistry Institute, Department of Common and Inorganic Chemistry, Astrakhanskaya 83, 410012 Saratov, Russia.
Goryacheva
I Yu
IY
Rusanova
T Yu
TY
Yurasov
N A
NA
Galve
R
R
Marco
M-P
MP
De Saeger
S
S
D000906:Antibodies / Q000276:immunology; D002733:Chlorophenols / Q000032:analysis* / Q000276:immunology; D005782:Gels / Q000737:chemistry; D007118:Immunoassay; D009793:Ochratoxins / Q000032:analysis* / Q000276:immunology; D014920:Wine / Q000032:analysis*
D000906:Antibodies; D002733:Chlorophenols; D005782:Gels; D009793:Ochratoxins; C025589:ochratoxin A; C024564:2,4,6-trichlorophenol