Abstract: | Abstract A reagentless fiber optic biosensor specific for NADH, associated with flow injection analysis and based on bacterial bioluminescence is described. Only a buffer solution as flowing stream is required for the probe which works without supply of its coreactants (FMN and N-decyl-aldehyde). These are coentrapped in a poly(vinyl)alcohol (PVA) matrix which allows their internal release in the vicinity of the immobilized enzymes. Two PVA matrices differing by the reticulation process have been tested : first, by polymerization with glutaraldehyde and secondly, by a cyclic freezing-thawing process. The self-containment working time was estimated at 1 and 1.5 h of continuous measurements, respectively. NADH was determined using flow injection analysis. The sensor gave excellent reproducibility (RSD ≤ 3%) in the linear dynamic range 5 — 500 pmol with an average cycle-time of 2.5 min. |