We studied the influence of graded degrees of hypertonic glucose or sucrose on the generation of oxygen-derived free radicals by rat neutrophils. Hypertonic glucose and sucrose exerted dose- and time-dependent inhibition of chemiluminescence amplified by luciferin analog (CLA-DCL) and luminol (L-DCL) in response to fMLP. Hypertonic glucose was more effective to this chemiluminescence inhibition than hypertonic sucrose was. This inhibition of hypertonicity was more effective in CLA-DCL than in L-DCL. Although the production of superoxide anion measured by the reduction of ferricytochrome c was more inhibited by hypertonic glucose than by hypertonic sucrose, the myeloperoxidase activity was not affected by either glucose or sucrose hyperosmolarity. These data suggest that hyperosmotic state by itself and an additional direct glucose-toxicity may contribute to the impaired neutrophil function in the diabetic state.