Abstract
A novel sandwich structured SiO 2 gel/cytochrome c (Cyt c)/SiO 2 gel was designed and constructed on conductive boron-doped diamond (BDD) film substrate. A SiO 2 gel membrane was first in situ deposited on the pretreated positive charged H-terminated BDD electrode with a simple and artful surface vapor sol–gel method. Cyt c was subsequently immobilized onto the SiO 2 membranes by electrostatic attraction, followed by another SiO 2 gel layer in situ depositing on it. The SiO 2 interlayer was conceived to play an important role in the resultant sandwich structured SiO 2/Cyt c/SiO 2/BDD electrode as a selective “semi-open” medium, which guaranteed the immobilized Cyt c to maintain high stability and perform good electrochemistry and biocatalysis responses. The bioactivity of Cyt c was well protected and the immobilized biomolecule even didn't denature at extremely high or low pH condition. More attractively, Cyt c in the sandwich structured electrode could be further oxidized into highly reactive Cyt c π-cation by two-step electrochemical oxidation, which could oxidize NO 2 − into NO 3 − in the solution. A sensitive determination approach of nitrite was accordingly built up based on this biocatalytic oxidative interaction for the first time and a possible mechanism of the interaction was herein proposed.
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