Sol–gel immobilization of acetylcholinesterase for the determination of organophosphate pesticides in olive oil with biosensors

This paper presents the construction of amperometric biosensors for the detection of organophosphorus insecticides widely used in the treatment of olive trees. The systems are based on the immobilisation of acetylcholinesterase on screen-printed electrodes by bioencapsulation in a sol–gel composite. The enzyme activity was estimated by measuring the thiocholine produced by the enzymatic hydrolysis of the acetylthiocholine chloride using cobalt phtalocyanine as mediator. The developed devices have been used to carry out inhibition studies with three pesticides: malathion, methidathion and dimethoate (in their oxidized form), and tested using standard solutions and real samples of olive oil. These biosensors showed good operational stability as they maintained their initial analytical signal response during 10 successive measurements, and a good reproducibility with a relative standard deviation of 3%. The limits of detection of the developed devices were very compatible with the maximum residue limit tolerated by international regulations, they were as low as 10−9 M for the widely used pesticide malaoxon(oxidized malathion). The developed sensors were successfully used for the determinations of insecticides in real samples of olive oil.