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Analytical and Bioanalytical Chemistry

, Volume 411, Issue 19, pp 4909–4917 | Cite as

A chiral unified chromatography–mass spectrometry method to analyze free amino acids

  • Adrien Raimbault
  • Magdalena Dorebska
  • Caroline WestEmail author
Research Paper
Part of the following topical collections:
  1. Young Investigators in (Bio-)Analytical Chemistry

Abstract

In this project, we aimed at analyzing native (or free) amino acids with supercritical fluid chromatography coupled to mass spectrometric detection, with modern instruments and methods, and maintaining as simple a mobile phase as possible to ensure applicability of the method. The purpose was twofold: (i) a generic method allowing for satisfactory elution of a wide range of amino acids (acidic, basic, or neutral residue) and (ii) resolution of the enantiomeric pairs. The Chiralpak ZWIX (+) and (−) stationary phases were selected as they are well-known for the enantioresolution of amino acids in liquid chromatographic modes. A wide range elution gradient, starting with a large concentration of carbon dioxide (90%) and finishing at 100% solvent (methanol containing 70 mM ammonium formate and 7% water) allowed the elution of 18 native proteinogenic amino acids out of 19 injected. In these conditions, enantioselectivity was achieved for 16 of them. The basic amino acids (arginine, histidine, and lysine) were the most difficult to elute in these conditions, resulting in rather poor peak shapes. Cysteine was never observed in any of the conditions tested. Sample application was attempted with two food supplements, tablets containing a mixture of 17 proteinogenic amino acids and capsules containing taurine and theanine that were not present in the standards used for the method development. The sample preparation method was very simple, involving dissolution of the tablets and capsules in acidified water, filtration, and dilution with methanol. Mass spectrometric detection (electrospray ionization with single-quadrupole mass detection) allowed for unambiguous identification of most amino acids, except for the leucine and isoleucine isomers that were not separated by the generic gradient. The observation of taurine and theanine also suggests that the method should be generally applicable to other native amino acids than the proteinogenic amino acids selected for the development of this method. As peak shapes and signal-to-noise ratios could still be improved, further developments are wanted to upgrade this method. Due to the wide gradient (10 to 100% co-solvent in carbon dioxide), the method cannot truly be called either supercritical fluid chromatography (SFC) or enhanced-fluidity liquid chromatography (EFLC), but should be related to “unified chromatography” (UC), joining SFC and HPLC.

Graphical abstract

Keywords

Amino acids Enhanced-fluidity liquid chromatography Food supplements Supercritical fluid chromatography Unified chromatography 

Notes

Acknowledgements

Adrien Raimbault is grateful for a PhD grant received from the Ministry of Higher Education and Research. Magdalena Dorebska received support from Erasmus program, from the Jagellonian University (Krakow, Poland) and the University of Orleans, France. Caroline West is grateful for the support received from the Institut Universitaire de France (IUF), of which she is a Junior Member. Waters Corporation is acknowledged for the support received through the Centers of Innovation program. Dr. Pilar Franco (Chiral Technologies) is acknowledged for the kind gift of Chiralpak ZWIX columns.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2019_1783_MOESM1_ESM.pdf (1 mb)
ESM 1 (PDF 1016 kb)

References

  1. 1.
    Maftouh M, Granier-Loyaux C, Chavana E, Marini J, Pradines A, Heyden YV, et al. Screening approach for chiral separation of pharmaceuticals: part III. Supercritical fluid chromatography for analysis and purification in drug discovery. J Chromatogr A. 2005;1088:67–81.  https://doi.org/10.1016/j.chroma.2004.12.038.CrossRefGoogle Scholar
  2. 2.
    Zhang Y, Wu D-R, Wang-Iverson DB, Tymiak AA. Enantioselective chromatography in drug discovery. Drug Discov Today. 2005;10:571–7.  https://doi.org/10.1016/S1359-6446(05)03407-0.CrossRefGoogle Scholar
  3. 3.
    Wang MZ, Klee MS, Yang SK. Achiral and chiral analysis of camazepam and metabolites by packed-column supercritical fluid chromatography. J Chromatogr B Biomed Sci Appl. 1995;665:139–46.  https://doi.org/10.1016/0378-4347(94)00502-V.CrossRefGoogle Scholar
  4. 4.
    West C. Enantioselective separations with supercritical fluids—review. Curr Anal Chem. 2014;10:99–120.  https://doi.org/10.2174/1573411011410010009.CrossRefGoogle Scholar
  5. 5.
    Lesellier E, West C. The many faces of packed column supercritical fluid chromatography—a critical review. J Chromatogr A. 2015;1382:2–46.  https://doi.org/10.1016/j.chroma.2014.12.083.CrossRefGoogle Scholar
  6. 6.
    West C. Current trends in supercritical fluid chromatography. Anal Bioanal Chem. 2018;410:6441–57.  https://doi.org/10.1007/s00216-018-1267-4.CrossRefGoogle Scholar
  7. 7.
    Olesik SV. Physicochemical properties of enhanced-fluidity liquid solvents. J Chromatogr A. 2004;1037:405–10.  https://doi.org/10.1016/j.chroma.2004.04.001.CrossRefGoogle Scholar
  8. 8.
    Chester TL. Peer reviewed: chromatography from the mobile-phase perspective. Anal Chem. 1997;69:165A–9A.  https://doi.org/10.1021/ac971559t.CrossRefGoogle Scholar
  9. 9.
    Taguchi K, Fukusaki E, Bamba T. Simultaneous analysis for water- and fat-soluble vitamins by a novel single chromatography technique unifying supercritical fluid chromatography and liquid chromatography. J Chromatogr A. 2014;1362:270–7.  https://doi.org/10.1016/j.chroma.2014.08.003.CrossRefGoogle Scholar
  10. 10.
    Desfontaine V, Losacco GL, Gagnebin Y, Pezzatti J, Farrell WP, González-Ruiz V, et al. Applicability of supercritical fluid chromatography–mass spectrometry to metabolomics. I—optimization of separation conditions for the simultaneous analysis of hydrophilic and lipophilic substances. J Chromatogr A. 2018;1562:96–107.  https://doi.org/10.1016/j.chroma.2018.05.055.CrossRefGoogle Scholar
  11. 11.
    West C, Melin J, Ansouri H, Mengue Metogo M. Unravelling the effects of mobile phase additives in supercritical fluid chromatography. Part I: polarity and acidity of the mobile phase. J Chromatogr A. 2017;1492:136–43.  https://doi.org/10.1016/j.chroma.2017.02.066.CrossRefGoogle Scholar
  12. 12.
    Camel V, Thiébaut D, Caude M, Dreux M. Packed column subcritical fluid chromatography of underivatized amino acids. J Chromatogr A. 1992;605:95–101.  https://doi.org/10.1016/0021-9673(92)85032-O.CrossRefGoogle Scholar
  13. 13.
    Wolrab D, Frühauf P, Gerner C. Direct coupling of supercritical fluid chromatography with tandem mass spectrometry for the analysis of amino acids and related compounds: comparing electrospray ionization and atmospheric pressure chemical ionization. Anal Chim Acta. 2017;981:106–15.  https://doi.org/10.1016/j.aca.2017.05.005.CrossRefGoogle Scholar
  14. 14.
    Sánchez-Hernández L, Bernal JL, del Nozal MJ, Toribio L. Chiral analysis of aromatic amino acids in food supplements using subcritical fluid chromatography and Chirobiotic T2 column. J Supercrit Fluids. 2016;107:519–25.  https://doi.org/10.1016/j.supflu.2015.06.027.CrossRefGoogle Scholar
  15. 15.
    Khater S, Canault B, Azzimani T, Bonnet P, West C. Thermodynamic enantioseparation behavior of phenylthiohydantoin-amino acid derivatives in supercritical fluid chromatography on polysaccharide chiral stationary phases. J Sep Sci. 2018;41:1450–9.  https://doi.org/10.1002/jssc.201701196.CrossRefGoogle Scholar
  16. 16.
    Wolrab D, Frühauf P, Gerner C, Kohout M, Lindner W. Consequences of transition from liquid chromatography to supercritical fluid chromatography on the overall performance of a chiral zwitterionic ion-exchanger. J Chromatogr A. 2017;1517:165–75.  https://doi.org/10.1016/j.chroma.2017.08.022.CrossRefGoogle Scholar
  17. 17.
    Vera CM, Shock D, Dennis GR, Farrell W, Shalliker RA. Comparing the selectivity and chiral separation of d- and l-fluorenylmethyloxycarbonyl chloride protected amino acids in analytical high performance liquid chromatography and supercritical fluid chromatography; evaluating throughput, economic and environmental impact. J Chromatogr A. 2017;1493:10–8.  https://doi.org/10.1016/j.chroma.2017.02.017.CrossRefGoogle Scholar
  18. 18.
    Ishibashi M, Ando T, Sakai M, Matsubara A, Uchikata T, Fukusaki E, et al. High-throughput simultaneous analysis of pesticides by supercritical fluid chromatography/tandem mass spectrometry. J Chromatogr A. 2012;1266:143–8.  https://doi.org/10.1016/j.chroma.2012.09.067.CrossRefGoogle Scholar
  19. 19.
    West C, Fougère L, Lesellier E. Combined supercritical fluid chromatographic tests to improve the classification of numerous stationary phases used in reversed-phase liquid chromatography. J Chromatogr A. 2008;1189:227–44.  https://doi.org/10.1016/j.chroma.2007.12.062.CrossRefGoogle Scholar
  20. 20.
    Zhang T, Holder E, Franco P, Lindner W. Method development and optimization on cinchona and chiral sulfonic acid–based zwitterionic stationary phases for enantiomer separations of free amino acids by high-performance liquid chromatography. J Chromatogr A. 2014;1363:191–9.  https://doi.org/10.1016/j.chroma.2014.06.012.CrossRefGoogle Scholar
  21. 21.
    Grecsó N, Forró E, Fülöp F, Péter A, Ilisz I, Lindner W. Combinatorial effects of the configuration of the cationic and the anionic chiral subunits of four zwitterionic chiral stationary phases leading to reversal of elution order of cyclic β3-amino acid enantiomers as ampholytic model compounds. Enantioseparations. 2016;1467:178–87.  https://doi.org/10.1016/j.chroma.2016.05.041.Google Scholar
  22. 22.
    Orosz T, Forró E, Fülöp F, Lindner W, Ilisz I, Péter A. Effects of N-methylation and amidination of cyclic β-amino acids on enantioselectivity and retention characteristics using cinchona alkaloid- and sulfonic acid-based chiral zwitterionic stationary phases. J Chromatogr A. 2018;1535:72–9.  https://doi.org/10.1016/j.chroma.2017.12.070.CrossRefGoogle Scholar
  23. 23.
    Lajkó G, Ilisz I, Tóth G, Fülöp F, Lindner W, Péter A. Application of cinchona alkaloid-based zwitterionic chiral stationary phases in supercritical fluid chromatography for the enantioseparation of Nα-protected proteinogenic amino acids. J Chromatogr A. 2015;1415:134–45.  https://doi.org/10.1016/j.chroma.2015.08.058.CrossRefGoogle Scholar
  24. 24.
    Lemasson E, Bertin S, Hennig P, Boiteux H, Lesellier E, West C. Development of an achiral supercritical fluid chromatography method with ultraviolet absorbance and mass spectrometric detection for impurity profiling of drug candidates. Part I: optimization of mobile phase composition. J Chromatogr A. 2015;1408:217–26.  https://doi.org/10.1016/j.chroma.2015.07.037.CrossRefGoogle Scholar
  25. 25.
    Akbal L, Hopfgartner G. Effects of liquid post-column addition in electrospray ionization performance in supercritical fluid chromatography–mass spectrometry. J Chromatogr A. 2017;1517:176–84.  https://doi.org/10.1016/j.chroma.2017.08.044.CrossRefGoogle Scholar
  26. 26.
    Desfontaine V, Tarafder A, Hill J, Fairchild J, Grand-Guillaume Perrenoud A, Veuthey J-L, et al. A systematic investigation of sample diluents in modern supercritical fluid chromatography. J Chromatogr A. 2017;1511:122–31.  https://doi.org/10.1016/j.chroma.2017.06.075.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Adrien Raimbault
    • 1
  • Magdalena Dorebska
    • 1
  • Caroline West
    • 1
    Email author
  1. 1.University of OrleansOrléansFrance

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