This study demonstrates the application of an online electrochemical system (OECS) as an in vivo method to investigate the dynamic change of microdialysate ascorbate in the olfactory bulb (OB) of rats during the acute period of olfactory dysfunction induced by intraperitoneal (i.p.) injection of 3-methylindole (3-MI). The OECS is developed by directly coupling an electrochemical detector to in vivo microdialysis for the direct monitoring of ascorbate. The system benefits from the good electrochemical activity of single-walled carbon nanotubes towards the oxidation of ascorbate and exhibits high selectivity, good stability, reproducibility and linearity for the measurement of ascorbate in the OB under physiological conditions. With this method, the basal level of microdialysate ascorbate in the OB is determined to be 48.64 +/- 5.44 mu M. The administration of 3-MI clearly increases the microdialysate ascorbate in the OB after 3-MI treatments and this increase is obviously alleviated by intravenous administration of ascorbate and glutathione (GSH) within 10 min after i.p. injection of 3-MI. These observations with the OECS suggest that ascorbate may be involved in chemical processes during the early stages of 3-MI-induced olfactory dysfunction. This study essentially validates the OECS as an in vivo method for effective measurement of ascorbate in the OB in rat brain and such a method will find interesting applications in investigating chemical process associated with ascorbate underlying olfactory dysfunction.