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Determination of D- and L-amino acids in biological samples by two-dimensional column liquid chromatography

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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 citrate-sodium chloride buffer. Enantioseparation is by subsequent injection of 3 µl 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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Author notes

  1. N. C. van de Merbel

    Present address: Pharma Bio-Research International B.V., P.O. Box 200, 9470 AE, Zuidlaren, The Netherlands

Authors and Affiliations

  1. Department of Analytical Chemistry, Free University, De Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands

    N. C. van de Merbel, H. Lingeman & U. A. Th. Brinkman

  2. Department of Applied Nutrition and Food Chemistry, University of Lund, P.O. Box 124, 22100, Lund, Sweden

    M. Stenberg & R. Öste

  3. Department of Analytical Chemistry, University of Lund, P.O. Box 124, 22100, Lund, Sweden

    G. Marko-Varga & L. Gorton

Authors
  1. N. C. van de Merbel
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  2. M. Stenberg
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  3. R. Öste
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  4. G. Marko-Varga
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  5. L. Gorton
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  6. H. Lingeman
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  7. U. A. Th. Brinkman
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van de Merbel, N.C., Stenberg, M., Öste, R. et al. Determination of D- and L-amino acids in biological samples by two-dimensional column liquid chromatography. Chromatographia 41, 6–14 (1995). https://doi.org/10.1007/BF02687993

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  • DOI: https://doi.org/10.1007/BF02687993

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