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5 different bio-tools have been developed for sensitive detection of three organophosphorus insecticides mainly used for the treatment of olive trees: Malaoxon, Omethoate and Methidathion-oxon. The systems are based on the immobilization of electric eel acetylcholinesterase on screen-printed electrode (SPE) and microtiter plates by 3 different methods of immobilization. These developed systems were optimized with laboratory samples and then tested on natural samples of olive oil after a simple liquid-liquid extraction. Among these five systems designed, the amperometric biosensor based on acetylcholinesterase immobilized by bioencapsulation in a sol-gel matrix presented the best performance in terms of operational stability, storage stability, reproducibility, and it proved to be best suited for the determination of insecticides in real samples of olive oil. This biosensor has been associated with a highly selective extraction method based on the use of Molecularly Imprinted Polymers (MIPs) to improve selectivity. The results showed that the association MIPs-biosensor can selectively detect and quantify the three organophosphorus insecticides in such a complex matrix as olive oil, with limits of detection much lower than the maximum residue limit tolerated by international regulations.

Enzymatic Biosensor Associated with Molecularly Imprinted Polymers for Sensitive and Selective Detection of Organophosphorus Insecticides in Olive Oil