1990 Fiscal Year Final Research Report Summary
Development of Specific and Sensitive Fluorogenic Reagents for Bioactive Amines and Their Applications
| Project/Area Number |
01571172
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Physical pharmacy
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| Research Institution | Hokkaido University |
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Principal Investigator |
MIURA Toshiaki Hokkaido University, College of Medical Technology, Associate Professor, 医寮技術短期大学部, 助教授 (10091505)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Bioactive amines / Specific fluorogenic reagents / Orthophthlaldehyde analogs / Histamine / Glutathione / Arginin / Ornithine decarboxylase / Flow injection analysis |
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| Research Abstract |
In this research project, specific, sensitive and rapid methods were established for the assay of several bioactive amines by using their specific fluorogenic reagents we had developed.
1. High-performance liquid chromatographic(HPLC) method was developed for the determination of putrescine in biological samples by using 1, 2-dibenzoyl-1, 4-dihydrobenzene as a precolumn fluores-cence labelling reagent. This HPLC method was shown to be applicable to the assay of ornithine decarboxylase(ODC) in rat livers.
2. By using 4-bromo-1, 2-benzenedicarbaldehyde(BBDA), a specific fluorogenic reagent for histamine, we developed a simple fluorometric method for the determination of histamine in biological samples. This reagent was applied to the flow injection analysis(FIA) method, which allows 40 samples to be assayed within 1 hour. The detection limit(S/N=5) of this FIA method for histamine was demonstrated to be 0.5 pmo1/injection.
HPLC method for assay of histamine was also developed by using 2-acetylbenzaldehyde(ABA) as a postcolumn fluorescence labelling reagent. This method was highly sensitive and applicable to the assay of histamine in tissue extract only after deproteinization.
3. Under the appropriate reaction conditions used ABA could be employed as a specific reagent for glutathione. Thus, this reagent was employed to the simple fluorometric method and the rapid FIA method for the assay of glutathione.
4. By using 2, 3-naphthalenedicarbaldehyde, a specific fluorogenic reagent for arginine, sensitive and rapid FIA method was developed for the assay of arginine in serum. Furthermore, 2, 5-dimethoxytetrahydrofuran was found to be specific fluorogenic reagent for tryptophane and applicable to the fluorometric method for the assay of tryptophane in amino acid mixtures.
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