Strategies for the improvement of an amperometric cholesterol biosensor based on electropolymerization in flow systems: use of charge-transfer mediators and platinization of the electrode.

Different configurations based on an amperometric biosensor with cholesterol oxidase entrapped in a polypyrrole film have been developed with a view to improving the analytical properties of this biosensor. The alternatives considered involve the simultaneous entrapment of the enzyme and a charge-transfer mediator as well as previous platinization of the surface of the Pt electrode. Both artificial (a ferrocene derivative) and natural (flavin nucleotides) mediators were studied as constituents of the charge-transfer process between the enzyme and the electrode. The comparative study of these biosensors, which were prepared in situ in a continuous flow system, made it possible to determine the advantages and disadvantages of each configuration when applied to flow-injection determination of cholesterol.