Sensitive two-dimensional determination of small amounts of D-amino acids in mammals and the study on their functions.
D-amino acids are the candidates of novel physiologically active substances and the marker molecules of diseases in mammals. In the present study, the two-dimensional determination of small amounts of D-amino acids in mammals has been performed after sensitive pre-column fluorescence derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). The two-dimensional HPLC system includes the isolation of the target amino acids as D+L mixtures using the micro-ODS column, and the determination of the enantiomers using the chiral column. This method enables the sensitive and selective determination of small amounts of D-amino acids in mammals without the interference of L-amino acids and peptides, and the presence and distribution of D-Leu, D-Ala, D-Pro, D-Thr and D-allo-Thr has been demonstrated in rats and mice. The regulation and the origins of these D-amino acids, and their relationships to the biological rhythms are also discussed.
Hamase
Kenji
K
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan. hamase@phar.kyushu-u.ac.jp
D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis* / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D051379:Mice; D010507:Periodicity; D051381:Rats; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D000596:Amino Acids; D000594:Amino Acid Oxidoreductases