THE APPLICATION OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY/DUAL ELECTROCHEMICAL DETECTOR: THE DETECTION OF BRAIN NEUROTRANSMITTERS

• Main Authors: TASI MING HSUN, 蔡明勳
• Other Authors: LIU TUNG SHENG
• Format: Others
• Language: zh-TW
• Published: 1994
• Online Access: http://ndltd.ncl.edu.tw/handle/03160099652409160312

碩士 === 東海大學 === 應用化學系 === 82 === High-performance liquid chromatography with dual electrochemical detector was applied to the simultaneous measurement of norephr- ine, epinephrine, dopamine, 3,4-dihydroxyphenyl acetic acid, homovanillic acid, 5-hydroxyindole acetic acid, 3-methoxytyramine , and 5-hydroxytryamine in mouse brain homogenate. In this study, an optimal reduction potential and redox ratio of all analytes were determined. This technique provides good resolution of these analystes and offers a selective detection on the basic of their retention behavior and electrochemical reversibilities. In addition,we found that the dopamine peaks in the hippocampus and locus coeruleus were contaminated whereas those in the striatum and substantia nigra were not. Hence, conventional HPLC with electrochemical detection assays in the hippocampus and locus coeruleus may lead to unreliable measurements. It can improved by determining dopamine from the cathodic chromatogram. The required brain tissue samples could be less than 4 mg. Iscratic separation of these analytes by a conventional ODS column was achieved within 15 mins, hence over 90 analyses could be performed in a single working day in our automatic HPLC system. Further improvement in detection limits and resolution can be made by using microbore column instead of conventional columns. This simple, efficient and reliable HPLC method can be used as a basic research tool for assaying biogenic amines and their metabolites in mouse brain homogenates.