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Amino Acids

, Volume 17, Issue 3, pp 293–300 | Cite as

Determination of the optical purity of threonine and hydroxyproline by capillary gas chromatography on a chiral stationary phase

  • B. Franssou
  • U. Ragnarsson
Full Papers

Summary

Experimental conditions for the derivatization and resolution by GLC of all stereoisomers of threonine and 4-hydroxyproline are reported. Threonine was in two steps converted toN,O-bisisobutoxycarbonyl 2,2,2-trifluoroethyl ester derivatives, the second of which was performed under anhydrous conditions. As such the enantiomers could pairwise be separated by capillary gas chromatography on a Chirasil-Val column. SinceL- andD-threonine eluted much earlier than the corresponding allo forms, quantitative determination of the allothreonine content inD- orL-threonine down to the one percent level could be simply accomplished but also enantiomeric impurities could be determined. Unlike for threonine, the corresponding 4-hydroxyproline isomers could not all be resolved asN,O-bisisobutoxycarbonyl 2,2,2-trifluoroethyl esters on this column. Although diastereomers could still be separated, the allo pair cochromatographed and the resolution for theL- andD-isomers was low. Complete separation of the 4-hydroxyproline isomers could be accomplished asN,O-bisprotected isobutyl amides, the formation of which required three derivatization steps. These were used for the determination of allohydroxyproline.

Keywords

Amino acids Allohydroxyproline Allothreonine Chiral separation Chirasil-Val Hydroxyproline Threonine 

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References

  1. Abe I. Fujimoto N, Nakahara T (1995a) 2,2,2-Trifluoroethyl chloroformate as a rapid derivatizing reagent of amino acids for fast enantiomer separation by gas chromatography. Chem Lett: 113–114Google Scholar
  2. Abe 1, Kawazuma M, Fujimoto N, Nakahara T (1995b)N-Alkyloxycarbonyl isobutylamides as readily prepared diamide derivatives of amino acids for separation of enantiomeric isomers by chiral phase capillary gas chromatography. Chem Lett: 329–330Google Scholar
  3. Elemes Y, Ragnarsson U (1996) Synthesis and spectroscopic characterization of derivatives of proteinogenic amino acids, simultaneously labelled with13C,15N and2H in the backbone. Chem Commun: 935–936 (and references therein)Google Scholar
  4. Frank H, Nicholson GJ, Bayer E (1977) Rapid gas chromatographic separation of amino acid enantiomers with a novel chiral stationary phase. J Chromatogr Sci 15: 174–176PubMedGoogle Scholar
  5. Fransson B, Ragnarsson U (1997) Determination of the optical purity for serine and threonine by gas Chromatography on a chiral column. Chem Lett: 779–780Google Scholar
  6. Gil-Av E, Feibush B, Charles-Sigler R (1966) Separation of enantiomers by gas liquid chromatography with an optically active stationary phase. Tetrahedron Lett: 1009–1015Google Scholar
  7. Husek P (1991) Rapid derivatization and gas chromatographic determination of amino acids. J Chromatogr 552: 289–299Google Scholar
  8. Husek P (1998) Chloroformates in gas chromatography as general purpose derivatizing agents. J Chromatogr B717: 57–91Google Scholar
  9. Koenig WA, Parr W, Lichtenstein HA, Bayer E, Oro J (1970) Gas chromatographic separation of amino acids and their enantiomers: Non-polar stationary phases and a new optically active phase. J Chromatogr Sci 8: 183–186Google Scholar
  10. König WA (1987) The practice of enantiomer separation by capillary gas chromatography. Huethig, HeidelbergGoogle Scholar
  11. Lloyd-Williams P, Sanchez A, Carulla N, Ochoa T, Giralt E (1997) Synthetic studies on threonines. The preparation of protected derivatives ofD-allo- andL-allo-threonine for peptide synthesis. Tetrahedron 53: 3369–3382Google Scholar
  12. Nakaparksin S, Birrell P, Gil-Av E, Oro J (1970) Gas chromatography with optically active stationary phases: resolution of amino acids. J Chromatogr Sci 8: 177–182Google Scholar
  13. Sutherland A, Willis CL (1997) Enantioselective syntheses ofα-amino-β-hydroxy acids, [15N]-L-allothreonine and [15N]-L-threonine. Tetrahedron Lett 38: 1837–1840Google Scholar

Copyright information

© Springer-Verlag 1999

Authors and Affiliations

  • B. Franssou
  • U. Ragnarsson
    • 1
  1. 1.Department of BiochemistryUniversity of UppsalaUppsalaSweden

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