1997 Fiscal Year Final Research Report Summary
Studies on the establishment of the analytical method for, and the distribution and change of the total D-amino acids in mammalian body
| Project/Area Number |
08457592
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Physical pharmacy
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| Research Institution | The University of Tokyo |
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Principal Investigator |
IMAI Kazuhiro The Univ.of Tokyo, Grad.School of Pharm.Sci., Professor, 薬学系研究科, 教授 (50012620)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUSHIMA Takashi The Univ.of Tokyo, Grad.School of Pharm.Sci., Research Associate, 薬学系研究科, 助手 (00272485)
SANTA Tomofumi The Univ.of Tokyo, Grad.School of Pharm.Sci., Research Associate, 薬学系研究科, 助手 (30187306)
HOMMA Hiroshi The Univ.of Tokyo, Grad.School of Pharm.Sci., Associate Professor, 薬学系研究科, 助教授 (50190278)
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| Project Period (FY) |
1996 – 1997
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| Keywords | D-aspartic acid / pineal gland / NBD-F / HPLC / chiral stationary phase / anti D-aspartate antibody |
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| Research Abstract |
During the course of our studies on the determination of D-amino acids in mammals, we found that a large amount (mMorder) of D-aspartic acid (D-Asp) is present in the rat pineal gland. The method we usedisa chiral stationary phase-HPLC separation adn fluorometoric detection of NBD-labelled amino acids. In the present study, to increase the efficiency of the enantioneric separation, we developed 3 new chiral stationary phase containing the same chiral moieties as the previous phases with an extended long alkyl chain from the aminopropylsilica support. One of them, APS-11-OA-2500S was the best for the enantiomeric separation of NBD-amino acids, amino acids ester and beta-amino butyricacid. The interaction between the analyte and the chiral moiety was also studied with NMR,showing some interaction of proton of amino moiety of amino acids and carbonyl moiety of the chiral stationary phase.
The whole body autoradiography of rat injected with ^<14>C labelled D-Asp showed the incorpration of radioactivity to the tissues where the endogenous D-Asp existed.
The polyclonal antibody to D-Asp revealed the localization of the endogenous D-Asp in the cytoplasm of pinealocyte which locate in the distal region of the gland. The specifid location of D-Asp was also observed in the pituitary gland, suggesting the exsitence of some important physiological roles of D-Asp in the mammaliam body.
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