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In this study, a direct competitive biomimetic enzyme-linked immune-sorbent assay (BELISA) based on a molecularly imprinted nanomembrane as an artificial antibody was developed for the determination of triazophos in real samples. The imprinted film was directly synthesized on the well surface of a 96-well plate using a dummy molecular template under the conditions of thermal polymerization. The developed BELISA using a hapten of triazophos as an enzyme-labeled probe is much more sensitive, simple, quick, steady and inexpensive than the other instrumental and immuno assay methods. Under optimal conditions, the linear range of the method was 0.001-10 000 μg L -1 with a good regression coefficient of 0.977. The sensitivity (IC 50 ) and the limit of detection (LOD) of BELISA were 428 μg L -1 and 0.001 μg L -1 , respectively. This method was performed to detect triazophos in cabbage and apple samples, and showed excellent recovery and relative standard deviations (RSDs) ranging from 70.5 to 119.8% and from 5.2 to 19.7%, respectively. The results correlated well with those obtained using high performance liquid chromatography-tandem mass spectrometry.

Biomimetic enzyme-linked immunoassay based on a molecularly imprinted 96-well plate for the determination of triazophos residues in real samples