Yang_2013_Anal.Biochem_437_144

A sensitive amperometric acetylcholinesterase (AChE) biosensor based on platinum nanoparticles (Pt NPs), carboxylic graphene (CGR), and nafion (NF)-modified glassy carbon electrode (GCE) has been developed. The Pt NPs-CGR-NF nanocomposites with excellent conductivity, catalysis, and biocompatibility offered an extremely hydrophilic surface for AChE adhesion. Chitosan (CS) was used as cross-linker to immobilize the AChE on Pt-CGR-NF-modified GCE. NF was used as a protective membrane of the AChE biosensors. The AChE biosensor showed favorable affinity to acetylthiocholine chloride (ATCl) and could catalyze the hydrolysis of ATCl with an apparent Michaelis-Menten constant value of 148muM. Under optimum conditions, the biosensor detected methyl parathion in the linear range from 1.0x10(-13) to 1x10(-10)M and from 1.0x10(-10) to 1x10(-8)M with a detection limit of 5x10(-14)M and detected carbofuran in the linear range from 1.0x10(-12) to 1x10(-10)M and from 1.0x10(-10) to 1x10(-8)M with a detection limit of 5x10(-13)M. The biosensor exhibited good sensitivity, acceptable stability, and reproducibility, thus providing a promising tool for analysis of enzyme inhibitors.