Chromatographia

, Volume 41, Issue 1–2, pp 6–14 | Cite as

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

  • N. C. van de Merbel
  • M. Stenberg
  • R. Öste
  • G. Marko-Varga
  • L. Gorton
  • H. Lingeman
  • U. A. Th. Brinkman
Originals

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 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.

Key Words

Column liquid chromatography Two-dimensional separation D- and L-amino acids Enantioseperation 

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Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1995

Authors and Affiliations

  • N. C. van de Merbel
    • 1
  • M. Stenberg
    • 2
  • R. Öste
    • 2
  • G. Marko-Varga
    • 3
  • L. Gorton
    • 3
  • H. Lingeman
    • 1
  • U. A. Th. Brinkman
    • 1
  1. 1.Department of Analytical ChemistryFree UniversityAmsterdamThe Netherlands
  2. 2.Department of Applied Nutrition and Food ChemistryUniversity of LundLundSweden
  3. 3.Department of Analytical ChemistryUniversity of LundLundSweden

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