Disposable multichannel immunosensors for 2,4‐dichlorophenoxyacetic acid using acetylcholinesterase as an enzyme label

Abstract

An improved version of the disposable multichannel immunochemical biosensor for the herbicide 2,4‐dichlorophenoxyacetic acid (2,4‐D) based on a screen‐printed amperometric transducer and monoclonal antibodies (MAb) against 2,4‐D is reported. Entrapment within a thin Nafion film was used for the direct immobilization of MAb at the electrode surface. The amount of the tracer (2,4‐D conjugated to acetylcholinesterase) bound in a competitive immunochemical reaction was determined amperometrically using acetylthiocholine iodide as substrate. The measuring procedure (times of incubation with tracer and substrate, pH, tracer concentration) was optimized. The sensor was able to detect less than 0.01 μg/L of free 2,4‐D in water. One analysis (8 samples) was completed in 30 min (20 min for immunochemical reaction, 5 min incubation with substrate, 5 min measurement). The performance of the immunosensor (two configurations) was evaluated on real samples (tap water) with added 2,4‐D. The determined amounts (mean values 0.097 to 0.105 and 0.89 to 1.13) corresponded well with the added contents of 2,4‐D (0.100 and 1.00 μg/L, respectively).