Fluorometric Determination of 1,2,3,4-Tetrahydro-6,7-dihydroxyisoquinoline in Biological Materials by HPLC
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概要
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In the belief that endogenous 1,2,3,4-tetrahydro-6,7-dihydroxyisoquinoline (DA-Fp) could be a potential marker involved in the etiology of various diseases such as Parkinsonism, we attempted to develop a fluorescence method for DA-Fp. It was synthesized by condensation of dopamine with formaldehyde according to an established method.Periodate was identified by screening from various oxidation reagents as a fluorescence reagent to DA-Fp. Optimal reaction conditions were obtained with 0.25 mM NaIO_4 in 0.1 M phosphate buffer (pH 8.0) at 37℃ for 15 min. The fluorescence spectrum of the derivative showed that we had found a new reaction specific for DA-Fp. This reaction we coupled on-line to high performance liquid chromatography (HPLC), which enabled us to achieve a highly sensitive method for determining DA-Fp. A working curve was linear from 2 to 800 pmol of DA-Fp per injection.To determine DA-Fp in biological materials, the pretreatment before HPLC was optimized by hydrolysis of its conjugate and suppression of the artifact with l-phenylephrine. Urinary excretion of DA-Fp in men was measured by this new present method. The urinary excretion of endogenous DA-Fp increased in a rabbit given L-DOPA. The DA-Fp concentration was determined in rat brain. The significance of DA-Fp in these biological materials is discussed and evaluated. We conclude that the present method will be useful for studying tetrahydroisoquinolines involved in meny diseases.
- 1997-11-15
著者
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Yoshioka Masanori
Faculty of Pharmaceutical Sciences, Setsunan University
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Shirahata Akira
Faculty Of Pharmaceutical Science Josai University
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Tamura Z
Faculty Of Pharmaceutical Sciences The University Of Tokyo
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Yoshioka Masanori
Faculty Of Pharmaceutical Sciences Setsunan University
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Shirahata Akira
Faculty Of Pharmaceutical Sciences Josai University
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TAMURA Zenzo
Faculty of Pharmaceutical Sciences, The University of Tokyo
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Tamura Zenzo
Faculty Of Pharmaceutical Sciences University Of Tokyo
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