Polyethyleneimine-coated reticulated vitreous carbon electrode with immobilized enzymes as a substrate detector Original Research Article

Polyethyleneimine (PEI) was covalently coupled to carbodiimide-activated reticulated vitreous carbon (RVC). The PEI-coated RVC was activated with glutaraldehyde, and glycerol dehydrogenase and diaphorase were then immobilized. The PEI-coated RVC with immobilized enzymes functioned both as an enzyme reactor and a working electrode in an amperometric detection system where NAD+/NADH was recycled by the immobilized enzymes. The coated RVC electrode showed good properties compared to uncoated RVC, such as a long lifetime and a constant response to a series of injections in a flow-injection system, resulting in a relative standard deviation of 1.4%. The calibration graph was linear from the detection limit 0.1 μM to 2 mM NADH in the absence of recycling.