[An application of electrochemical detector to dental pharmacology].

Abstract

The electrochemical detection (ECD) is highly sensitive and can distinguish selectively electrochemically active materials. A sensitive and selective ECD is widely used for high-performance liquid chromatography (HPLC) of phenolic compounds, such as catecholamines, indoleamines and phenolic steroids in biological materials. In the present study, the HPLC/ECD system was applied for the simultaneous determination of various dental drugs, such as phenol, guaiacol, m-cresol, o-chlorophenol, p-chlorophenol, m-chlorophenol, eugenol, pyrogallol and resorcin. As a compromise between sensitivity and stability, the applied voltage was set at 1.2 V vs the reference electrode. These drugs were rapidly and selectively separated on a reversed-phase column using 10 mM acetate buffer (pH 4.0)-methanol (7:3, v/v) as a mobile phase. The detection limits of these drugs were determined to be 0.25 pmol per injection. These results imply that the HPLC/ECD method should be capable of determining these dental drugs with high sensitivity. The ECD was also applied to determine the enzyme activity. A highly sensitive and specific assay for alkaline and acid phosphatases and N-acetyl-beta-glucosaminidase in biological materials, such as plasma, saliva and bone, has been established. Phenol, formed enzymatically from the substrate, was determined by HPLC/ECD. The retention time of phenol was 7 min and no other peaks were observed. The method is rapid and sensitive with a detection limit for phenol of as little as 0.5 pmol. Thus, as little as 0.5 microliters of rat plasma or 10 microliters of human saliva is all that is required for both alkaline and acid phosphatase assays. The assay is accurate and reproducible. Using this assay, alkaline and acid phosphatase activities in human saliva and in rat saliva elicited by pilocarpine and isoproterenol were determined. It will be possible to apply this new assay method to extremely small biological samples. Thus, the application of the HPLC/ECD system to determine dental drugs and enzyme activity with high sensitivity should prove to be of great value in the field of dental pharmacology.