Development of a disposable mimetic biosensor for organophosphorus and carbamate pesticides aiming quality control of water supply

Acetylcholinesterase enzyme (AChE) has been widely applied in biosensor development for pesticide detection. However, the application of those biosensors is limited due to their lack of robustness associated with low enzymatic stability and AChE high cost. An alternative to enzymes, it is the use of synthetic molecules which mimic enzymatic behavior, besides its low cost and superior stability, those are classified as biomimetic molecules. In this work is presented three potential catalysts: [4-(1E)ethanehydrazonoyl]benzoic acid, pralidoxime iodide (2-PAM), and polyhydroxamicalkalnoate polymer (PHA) with alpha nucleophile functional groups, such as hydrazone, oxime, and hydroxamic acid, respectively. The aforementioned molecules were evaluated based on two criteria for classification as AChE biomimetic: 1) catalysis of acetylthiocholine substrate (ATCh) hydrolysis and 2) interaction with organophosphorus pesticide. Spectrophotometrical Ellman´s method revealed that PHA polymer was the most efficient one, since its second order rate constant was 13867 mol-1 L s-1, which was 5 times superior than 2-PAM one and 232 times higher than hydrazone. Then, each molecule was analyzed through spectrophotometric method to verify the interaction with paraoxon organophosphorus pesticide. Only the PHA exhibited such interaction in the studied conditions. Theoretical calculations showed how the approach between PHA and ATCh, and also with paraoxon occurred. Regarding the obtained results, PHA was the chosen biomimetic molecule to be applied in the electrochemical sensor for organophosphorus detection. First of all, PHA was immobilized on the surface of disposable screen printed electrode through blend formation with polyethylene glycol methyl ether (mpEG), avoiding PHA dissolution. The oxidation of thiocholine generated towards ATCh catalysis performed by immobilized PHA was verified at 0.46 V vs Ag/AgCl by differential pulse voltammetry. The linear equation obtained through calibration curve for paraoxon concentration range of 1.0 to 10.0 µmol L-1 was I(%) = [4.8x106 C (mol L-1) + 0.8169]×100 (R2 = 0.99969), with limit of detection of 3.63x10-7 mol L-1. Water supply samples were analyzed considering the added paraoxon concentrations of 3.0; 6.0 e 8.0 µmol L-1, recovering 115%; 106% and 94.5%, respectively. The presented results showed that a biomimetic electrochemical sensor was successfully developed for detection of paraoxon organophosphorus pesticide. (AU)