| Project/Area Number |
62570972
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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 | Kitasato University |
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Principal Investigator |
KINOSHITA Toshio School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (70053816)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Junko School of Pharmaceutical Sciences, Research Assistant, 薬学部, 助手 (60220646)
NIMURA Noriyuki School of Pharmaceutical Sciences, Assistant Professor, 薬学部, 講師 (50118832)
平賀 やよい 北里大学, 薬学部, 助手 (80050706)
加藤 武彦 北里大学, 薬学部, 助手 (50050589)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | amino acid / protein / peptide / amino acid analysis / fluorescence analysis / high performance liquid chromatography / o-phthalaldehyde / post-column detection / 蛋白質同定 / 高速液体クロマトグラフィー(HPLC) / o^-フタルアルデヒド / 高速液体クロマトグラフィー / O-フタルアルデヒド / ポストカラム反応 / イミノ酸 / プロリン |
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| Research Abstract |
This research project developed a high intelligent amino acidanalyzer consisted of following components. (1) Separation system; Amino acid separation was based on a reversed-phase ion-pair high performance liquid chromatography using a octadecylsilyl silica gel column with aqueous solution of sodium dodecyl sulfate as a mobile phase. In this separation system, 17 protein amino acids and ammonia were completely resolved within only 12 min. The analyzing time was 4 to 5 times shorter than conventional ion-exchange separation. (2) Detection system; Amino acids eluted from the above separation system were converted to fluorescence compounds in a online reaction system. The fluorescence reaction comprised of two steps, oxidative cleavage of imino acids such as proline to primary amino compounds by sodium hypochlorite, and fluorescence development with o- phthalaldehyde/N-acetyl-L-cysteine reagent. The use of this system facilitated the detection of imino acids at the same concentration level as that of amino acids. The detection limit for all imino and amino acids was a few picomoles. (3) Data processing system; By use of computer aided data processing system, protein samples were directly identified. The computer program contained a processing part for chromatgraphic data and calculating part for correlation factor based on amino acid contents of known proteins.
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