Abstract
Gas chromatography (GC) is a commonly used technique in amino acid analysis (AAA). However, one of the requirements of the application of GC for AAA is a need for the polar analytes to be converted into their volatile, thermally stable derivatives. In the last two decades, alkyl chloroformates have become attractive derivatization reagents. The reagents react immediately with most amino acid functional groups in aqueous matrices and the process can easily be coupled with liquid–liquid extraction of the resulting less-polar derivatives into immiscible organic phase. Here, we describe a simple protocol for in situ derivatization of amino acids with heptafluorobutyl chloroformate followed by subsequent chiral as well as nonchiral GC/mass spectrometric analysis on a respective nonpolar fused silica and an enantioselective Chirasil-Val capillary column.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gehrke CW, Kuo CT, Zumwalt R W (1987) Amino acid analysis by gas chromatography. Vol. I-III CRC Press, Boca Raton
Hušek P, Šimek P (2001) Advances in amino acid analysis. LC-GC North America 18: 986–999
Molnar-Perl I (2005) Quantitation of amino acids and amines by chromatography. J Chromatogr–Library, Vol. 70, Elsevier, Amsterdam
Hušek P, Šimek P (2001) Alkyl chloroformates in sample derivatization strategies for GC analysis. Current Pharm Anal 2:21–41
Hušek P et al (2008) Fluoroalkyl chloroformates in treating amino acids for gas chromatographic analysis. J Chromatogr A 1186:391–400
Šimek P, Hušek P, Zahradníčková H (2008) Gas chromatographic-mass spectrometric analysis of biomarkers related to folate and cobalamin status in human serum after dimercaptopropane-sulfonate reduction and heptafluorobutyl chloroformate derivatization. Anal Chem 80:5776–5782
Patzold R, Bruckner H (2005) Chiral separations of amino acids. In: Molnar-Perl I (ed.) Quantitation of amino acids and amines by chromatography. J Chromatogr–Library vol. 70, Elsevier, Amsterdam
Zahradníčková H, Hartvich P, Šimek P, Hušek P (2008) Gas chromatographic analysis of amino acid enantiomers in Carbetocin peptide hydrolysates after fast derivatization with pentafluoropropyl chloroformate. Amino Acids 35: 445–450
Zahradníčková H, Hušek P and Šimek P (2009) GC separation of amino acid enantiomers via derivatization with heptafluorobutyl chloroformate and Chirasil-L-Val column. J Sep Sci 25:3919–3924
Acknowledgments
This work was supported by the Fund for Research Support, EEA/Norway grant No. A/CZ0046/1/0018, the Czech Science Foundation, project No.213/09/2014, and the Internal Grant Agency of Ministry of Health of the Czech Republic, No. NS9755-4/2008.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Šimek, P., Hušek, P., Zahradníčková, H. (2012). Heptafluorobutyl Chloroformate-Based Sample Preparation Protocol for Chiral and Nonchiral Amino Acid Analysis by Gas Chromatography. In: Alterman, M., Hunziker, P. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 828. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-445-2_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-445-2_13
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-444-5
Online ISBN: 978-1-61779-445-2
eBook Packages: Springer Protocols