Acetylthiocholine/acetylcholine and thiocholine/choline electrochemical biosensors/sensors based on an organically modified sol–gel glass enzyme reactor and graphite paste electrode

Abstract

Electrochemical sensors for acetylthiocholine and acetylcholine are described. The non-mediated electrochemistry of acetylthiocholine and thiocholine is studied on the surface of graphite paste electrode and results show that acetylthiocholine is directly oxidized/reduced at >0.32 V vs. Ag/AgCl in both acidic and basic medium. In basic medium, both cathodic and anodic peak currents are less as compared to that of the same amount in acidic medium, which shows that the kinetics of non-enzymatic hydrolysis of acetylcholine into electroactive thiocholine is faster in acidic medium and slower in basic medium. Thiocholine is directly oxidized/reduced at >0.35 V vs. Ag/AgCl with relatively larger anodic current compared to cathodic peak current similar to that of acetylcholine results recorded in acidic medium (pH 6.0). The electrochemical sensor/biosensors for acetylthiocholine/acetylcholine and thiocholine/choline are developed using two enzyme reactors: (1) acetylcholinesterase (AChE) encapsulated organically modified sol–gel glass, and (2) choline oxidase (ChO) immobilized within mediators (tetracyanoquinodimethane (TCNQ), tetrathiafulvalene (TTF), and dimethyl ferrocene (dmFc))-modified graphite paste electrodes. The AChE-immobilized into organically modified sol–gel glass behaves as the reactor for enzymatic hydrolysis of acetylthiocholine/acetylcholine into thiocholine/choline, whereas mediator- and ChO-modified paste electrodes are used for the detection of thiocholine/choline through mediated mechanism. The electrochemistry of AChE-generated thiocholine is studied at the mediator-modified electrodes in the presence and absence of ChO. It is observed that thiocholine undergoes both mediated and non-mediated oxidation in the absence of ChO as well as oxidation through enzyme-catalyzed mediated reactions. The results based on cyclic voltammetry on the oxidation of thiocholine at the surface of mediator-modified electrodes in the presence and absence of ChO are reported. In the presence of the ChO large anodic current is observed near the mediator's redox potentials as compared to the anodic current in the absence of enzyme, which shows mediated bioelectrochemistry of thiocholine. The typical response curves for the detection of thiocholine/choline using mediators and ChO-modified electrodes below 0.24 V vs. Ag/AgCl in 0.1 M Tris–HCl buffer pH 8.0 are reported. Comparative analytical performance on the mediated electrochemical responses of the biosensors is discussed.