Composite carbon paste biosensor for phenolic derivatives based on in situ electrogenerated polypyrrole binder

Amperometric biosensors based on new composite carbon paste (CPE) electrodes have been designed for the determination of phenolic compounds. The composite CPEs were prepared by in situ generation of polypyrrole (PPy) within a paste containing the enzyme polyphenol oxidase (PPO). The best paste composition (enzyme/pyrrole monomer/carbon particles/Nujol) was determined for a model enzyme, glucose oxidase, according to the enzymatic activity of the resulting electrodes and to the enzyme leakage from the paste during storage in phosphate buffer. The in situ electrogenerated PPy enables improvement in enzyme immobilization within the paste since practically no enzyme was lost in solution after 72 h of immersion. Moreover, the enzyme activity remains particularly stable under storage since the biocomposite structure maintains 80% of its activity after 1-month storage. Following the optimization of the paste composition, PPO-based carbon paste biosensors were prepared and presented excellent analytical properties toward catechol detection with a sensitivity of 4.7 A M(-1) cm(-2) and a response time lower than 20 s. The resulting biosensors were finally applied to the determination of epicatechin and ferulic acid as flavonol and polyphenol model, respectively.