Abstract A novel approach to the design and use of an air-segmented flow injection (ASFI) system is described. The ASFI system possesses features of both classical flow injection (FI) and air-segmented continuous flow (ASCF) methods. The entire air-segmented sample zone is injected through a simple 6-way valve into the carrier stream. Preliminary studies of dye dispersion and spectrophotometric glucose determinations in wine and serum samples are presented. These results indicate that the proposed system has advantages for most determinations in sensitivity, sampling rate and solution consumption when compared to the other flow analysis techniques. Analytical results are also compared with those obtained by kinetic-based determinations.