A sandwich structured SiO(2)/cytochrome c/SiO(2) on a boron-doped diamond film electrode as an electrochemical nitrite biosensor.

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 pi-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.