Abstract.The chemiluminescence reaction of lucigenin with glucose and fructose in the presence of Brij-35 and in alkaline solution was used to develop kinetic methods for the determination of both hexoses individually and mixed together. A stopped-flow mixing module coupled to a photomultiplier tube allowed kinetic and equilibrium measurements, which were used for the individual determination of glucose and fructose. The same measurements were used to analyse the mixture of both sugars using a two equation system, which was resolved by multiple linear regression. The ranges of the calibration graphs obtained by using kinetic and equilibrium measurements were 251000gml1 and 751100gml1, respectively, for glucose and 1350gml1 and 5250gml1, respectively, for fructose. Mixtures of glucose and fructose in ratios between 1:0.4 and 40:1 were satisfactorily resolved.