New microdialysis-electrochemical device for simultaneous determination of ascorbic acid and 5-hydroxyindole-3-acetic acid in rat striatum

Abstract

A new device combining microdialysis with electrochemical microsensor was developed. It can be applied to monitor the biomolecules in the brain for biological and pharmaceutical research. In this paper, the device was applied to simultaneously determine ascorbic acid (AA) and 5-hydroxyindole-3-acetic acid (5-HIAA) in rat striatum. The microsensor used for the device was poly (sulphosalicylic acid) microsensor, which exhibited a good electrocatalytic effect on oxidization of AA and 5-HIAA. The oxidation currents measured by differential pulse voltammetry (DPV) were linear for AA in the range of 0.02–1.0 mmol l −1, and for 5-HIAA from 0.5 to 10.0 μmol l −1 ( r=0.9998 and 0.9991, respectively). The detection limits were calculated to be 0.01 mmol l −1 for AA and 0.25 μmol l −1for 5-HIAA (S/N=3). Studies also showed that co-existing substances in biological fluids did not interfere with AA and 5-HIAA determination when using this microsensor. Since, the substances in the microdialysate are easily oxidized by air, the microdialysate in this device was under the protection of N 2. It was found that the concentrations of AA and 5-HIAA in rat striatum were 215±5 and 6.21±0.61 μmol l −1 (mean±S.E.M., n=7), respectively with this device under the protection of N 2. In addition, the effect of the nitric oxide donor, sodium nitroprusside (SNP), on 5-HIAA in the rat striatum was investigated. It was found that a high concentration of SNP (1.0 mmol l −1) resulted in a 34% increase in 5-HIAA level.