Quaternary ammonium functionalized clay film electrodes modified with polyphenol oxidase for the sensitive detection of catechol.
Naturally occurring Cameroonian smectite clay has been grafted with trimethylpropylammonium (TMPA) groups and the resulting organoclay has been deposited onto a glassy carbon electrode surface as a suitable immobilization matrix for polyphenol oxidase (PPO). High sensitivity of the electrochemical device to catechol biosensing can be achieved when the enzyme was impregnated within the organoclay film subsequent to its deposition due to favorable electrostatic interaction between PPO and the TMPA-clay layer. The bioelectrode preparation method was also compatible with the use of a mediator (i.e., ferrocene) and the best performance was obtained with a three-layer configuration made of glassy carbon coated with a first layer of ferrocene (Fc), which was then covered with the PPO-impregnated TMPA-clay layer, and finally overcoated with an enzyme-free TMPA-clay film acting as a protecting overlayer to avoid leaching of the biomolecule in solution. The electrochemical behavior of the modified film electrodes was first characterized by cyclic voltammetry and, then, they were evaluated for the amperometric biosensing of the model analyte catechol in batch conditions and in flow injection analysis. Various experimental parameters likely to influence the biosensor response have been investigated, including the electrode preparation mode (composition configuration, thickness), the usefulness of a mediator, the operating potential and pH of the medium, as well as the advantageous features of the TMPA-clay in comparison to related film electrodes based on non-functionalized clays. The organoclay was found to provide a favorable environment to enzyme activity and the multilayer configuration of the film electrode to provide a biosensor with good characteristics, such as an extended linear range for catechol detection (2 x 10(-8) to 1.2 x 10(-5)M) and a detection limit in the nanomolar range (9 x 10(-9)M).
Mbouguen
Justin Kemmegne
JK
Laboratoire de Chimie Physique et Microbiologie pour l'Environnement, Unité Mixte de Recherche UMR 7564, CNRS, Nancy-Université, 405, rue de Vandoeuvre, F-54600 Villers-les-Nancy, France.
Ngameni
Emmanuel
E
Walcarius
Alain
A
D000538:Aluminum Silicates / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004156:Catechol Oxidase / Q000737:chemistry*; D002396:Catechols / Q000032:analysis*; D000077215:Clay; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008567:Membranes, Artificial; D008839:Microelectrodes; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000538:Aluminum Silicates; D002396:Catechols; D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D000644:Quaternary Ammonium Compounds; C520881:trimethylpropylammonium; D004156:Catechol Oxidase; C034221:catechol; D000077215:Clay