Determination of D- and L-amino acids in biological samples by two-dimensional column liquid chromatography |
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| Authors: | N. C. van de Merbel M. Stenberg R. Öste G. Marko-Varga L. Gorton H. Lingeman U. A. Th. Brinkman |
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| Affiliation: | (1) Department of Analytical Chemistry, Free University, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands;(2) Department of Applied Nutrition and Food Chemistry, University of Lund, P.O. Box 124, 22100 Lund, Sweden;(3) Department of Analytical Chemistry, University of Lund, P.O. Box 124, 22100 Lund, Sweden;(4) Present address: Pharma Bio-Research International B.V., P.O. Box 200, 9470 AE Zuidlaren, The Netherlands |
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| Abstract: | Summary A two-dimensional, column liquid chromatographic system is used for the determination of the D- and L-enantiomers of amino acids in biological samples. Separation of the amino acids is first on ion-exchange column by gradient elution with a sodium citratesodium chloride buffer. Enantioseparation is by subsequent injection of 3  l heart-cuts of the individual amino acids onto a second column with a chiral crown ether stationary phase. Finally, fluorescence detection is after post-column labelling of the amino acids using ano-phthalaldehyde-2-mercaptoethanol reagent solution. The high separation power and selectivity of the system allow processing of complex samples with hardly any additional treatment and the determination of small quantities of D-amino acids in the presence of excess L-form. Applicability of the system is illustrated by the determination of D- and L-aspartate, serine, glutamate and alanine in various complex biological samples, such as protein hydrolysates, urine and biotechnological and food samples. Data are given on detectability, repeatability and linearity. |
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| Keywords: | Column liquid chromatography Two-dimensional separation D- and L-amino acids Enantioseperation |
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