| Summary: | 碩士 === 國立交通大學 === 應用化學系 === 91 === Analysis of biogenic amines by Micellar Electrokinetic Chromatography with Laser-Induced Fluorescence
Student：Yi-Fang Chen Advisor： You-Zung Hsieh
Department of Applied Chemistry
National Chiao Tung University
Abstract
Mono- or polyamines involved in biological activities and that exist in nature are sometimes called by biogenic amines. Some of them are biologically synthesized from amino acids by amino acid decarboxylase degradation and play an important role in living systems. In this study, analysis of fourteen biogenic amines by micellar electrokinetic capillary electrophoresis and laser induce fluorescence were investigated. The selected derivatizing reagent was fluorescein isothiocyanate ( FITC).
In order to achieve an optimum detection sensitivity and separation efficiencies,it was necessary to optimize precolumn derivatization conditions with respect to reaction time, pH, buffer concentration and the molar ratios of FITC to biogenic amines. It must also study the optimum separation conditions. In an optimized condition of micellar electrokinetic chromatography, using 40 mM sodium borate buffer (pH 10.1) with 40 mM sodium dodecyl sufate (SDS), fourteen biogenic amines could be successful separated.
To check the applicability of the method, affective disorder patient’s urine was analyzed. We additional spiked E and NE to identify the existence of E and NE in the urine. Qualitatively analysis, we can find seven kinds of biogenic amines in urines (Put, Oct, 5-HT, E, NE, Dop, Agm).Quantitatively analysis, we can find the different urinary catecholamine amounts of normal people and the patients. Another application in the beers and mastitis milk, we had also found five kinds of biogenic amines in beers (Put, Him, Tyr, PEA, Agm) and one kind of biogenic amines in milk (Put).
In the capillary electrophoresis microchip, we could successfully separate five kinds of biogenic amines (Put, Him, 5-HT, Dop, Agm) within 3 minutes. The optimum running buffer was pH 10.1,20 mM Borate/20 mM SDS.
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