Amperometric biosensor with HRP immobilized on a sandwiched nano-Au / polymerized m-phenylenediamine film and ferrocene mediator.

An amperometric biosensor has been developed for the determination of H(2)O(2) in plant samples. Horseradish peroxidase (HRP) is immobilized on a sandwiched nano-Au particle / m-phenylenediamine polymer film by glutaraldehyde cross-linking. The film is formulated on the carbon paste electrode (CPE) blended with ferrocene as an electron transfer mediator. On the low concentration range, the current response is related to the H(2)O(2) concentration linearly from 0 to 8x10(-6) M with a detection limit of 1.3x10(-7) M. On a wider concentration range of 8x10(-6) to 1.4x10(-4) M, the reciprocal of current response is linearly related to the reciprocal of H(2)O(2) concentration. The apparent Michaelis-Menten constant (K(m)(app)) was calculated to be 0.0334 mM. The sensor has been tested by determining H(2)O(2) concentration in plant leaf samples.