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 0.1μM to 2mM NADH in the absence of recycling. Copyright (C) 1998 Elsevier Science B.V.
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