Determination of D- and L-amino acids in mouse kidney by high-performance liquid chromatography.
A method for the enantiomeric analysis of amino acids of mammalian tissues is described. An excellent resolution of D- and L-enantiomers of common protein amino acids was achieved by employing a combination of thin-layer chromatography and high-performance liquid chromatography. D-Enantiomers and L-enantiomers of glutamate, aspartate, glutamine, asparagine, serine, threonine, alanine, proline, tyrosine, valine, methionine, isoleucine, leucine, phenylalanine and histidine, as well as glycine were derivatized with 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide. The amino acid diastereomers were separated by two-dimensional thin-layer chromatography. Each amino acid diastereomer was then analysed by reversed-phase high-performance liquid chromatography for the resolution of D- and L-enantiomers. Very sharp peaks were obtained using a conventional octadecylsilyl-bonded column, and the possibility of analysing these amino acids (except tyrosine and histidine) in subnanomole amounts was indicated. The method was used to demonstrate the presence of D-enantiomers of alanine, proline and serine in mouse kidney.
Nagata
Y
Y
Department of Life Science, Himeji Institute of Technology, Japan.
Yamamoto
K
K
Shimojo
T
T
D000409:Alanine / Q000032:analysis; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods; D007668:Kidney / Q000737:chemistry*; D051379:Mice; D011392:Proline / Q000032:analysis; D012694:Serine / Q000032:analysis; D013237:Stereoisomerism
D000596:Amino Acids; D012694:Serine; D011392:Proline; D000409:Alanine