Determination ofD-amino acids. II. Use of a bifunctional reagent, 1,5-difluoro-2,4-dinitrobenzene (original) (raw)

Abstract

1-fluoro-2,4-dinitrophenyl-5-l-alanine amide has been synthesized in high yield (76%) from 1,5-difluoro-2,4-dinitrobenzene andl-Ala-NH2. This compound contains a reactive fluorine atom which can be used for the reaction with a mixture ofl- andd-amino acids. The resulting diastereomers which are obtained in quantitative yield can be separated and estimated by HPLC. With the five amino acids studied (Ala, Asp, Glu, Met and Phe),l-diastereomers were eluted from the reverse-phase column befored-diastereomers. This behavior can be explained by a stronger intramolecular hydrogen bonding in the latter diastereomer. When artificial mixtures of the five amino acids containing known proportions ofl- andd-isomers were derivatized with the reagent and the reaction products analyzed by HPLC, it was possible to determine the relative content of each isomer in nanomole range.

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Abbreviations

DMSO:

dimethylsulfoxide

DNDEAP-a a:

N-(2,4-dinitro-5-diethylaminophenyl)-amino acid

DNP:

2,4-dinitrophenyl

FDAA=F-DNP-l-Ala-NH2 :

1-fluoro-2,4-dinitrophenyl-5-l-alanine amide

FFDNB:

1,5-difluoro-2,4-dinitrobenzene

HO-DAA:

1-hydroxy-2,4-dinitrophenyl-5-l-alanine amide

HPLC:

high performance liquid chromatography

l-Ala-NH2-HCl:

l-Alanine amide hydrochloride

TEAP:

triethylammonium phosphate

TLC:

thin-layer chromatography

References

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Author notes

1. Peter Marfey
   Present address: Department of Biological Sciences, State University of New York at Albany, 12222, Albany, N.Y., USA

Authors and Affiliations

1. Department of Chemistry, Carlsberg Laboratory, Gamble Carlsberg Vej 10, DK-2500, Copenhagen Valby
   Peter Marfey

Additional information

Other abbreviations are according to the guideline of the IUPAC-IUB. Commission of Biochemical Nomenclature.

Accepted by:K. Klenow. E. Lund andS.O. Andersen

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Marfey, P. Determination ofD-amino acids. II. Use of a bifunctional reagent, 1,5-difluoro-2,4-dinitrobenzene.Carlsberg Res. Commun. 49, 591 (1984). https://doi.org/10.1007/BF02908688

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• DOI: https://doi.org/10.1007/BF02908688

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