| Summary: | 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.
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