Abstract
In this paper, two acetylcholinesterase (AChE) immobilization methods on the surface of a glassy carbon electrode (GCE) were compared. AChE-employed homogenate from chicken brain was home-made in the laboratory. The immobilization methods employed a cross-linking method with glutaraldehyde as a cross-linking agent, bovine serum albumin (BSA) as a protectant, and sol-gel method with tetraethoxysilane (TEOS). AChE was immobilized on chitosan membranes by these two immobilization methods. Then the enzyme membrane was fixed on the surface of GCE to prepare an AChE-GCE amperometric biosensor for the detection of organophosphorus pesticide. Measuring the activity of immobilization AChE by the oxidation current of thiocholine (TCh), produced by hydrolysis of the acetylthiocholine iodide (ATChI) substrate, the results showed that the activity of AChE-employed sol-gel method with TEOS was higher than the cross-linking method with glutaraldehyde. Biosensor sensitivity and reproducibility were statistically evaluated for the different immobilization methods and the most suitable immobilization procedure was selected.
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