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The development of a four-band capillary optical immunosensor for the simultaneous determination of mesotrione, hexaconazole, paraquat, and diquat is described. Four distinct bands (each corresponding to a different analyte) are created in the internal walls of a plastic capillary by immobilizing protein conjugates of the analytes. To perform the assay, the capillary is filled with a mixture of anti-analyte-specific antibodies together with a standard or sample containing the analyte(s). After a short incubation, a mixture of the appropriate second antibodies labeled with fluorescein is introduced into the capillary. To measure the fluorescence intensity bound onto each band, the capillary was scanned, perpendicularly to its axis, by a laser light beam. Part of the emitted photons were trapped into the capillary walls and waveguided to a photomultiplier placed at the one end of the capillary. The analytical characteristics of the assays of mesotrione, paraquat, diquat, and hexaconazole were as follows: detection limits of 0.04, 0.06, 0.09, and 0.10 ng/mL, respectively; dynamic ranges up to 9, 6, 12, and 15 ng/ mL, respectively, intra- and interassay CVs less than 10%. The analytical characteristics of the assays were comparable with those of the corresponding single-analyte fluoroimmunoassays performed in microtitration wells, proving the ability of the proposed immunosensor for reliable multianalyte determinations. Moreover, the combination of low-cost disposable plastic capillary tubes with the low consumption of reagents, the short assay time, and the multianalyte feature of the proposed immunosensor indicates its potential for environmental analysis.

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