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Free Amino Acid Analysis in Honey Samples by Hydrophilic Interaction Liquid Chromatography with UV Detection Using Precolumn Derivatization with Dansyl Chloride

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Abstract

A new hydrophilic interaction chromatography (HILIC) method was developed for free amino acid analysis in honey samples after pre-column derivatization of amino acids with dansyl chloride (Dns-Cl). This method consisted of the sample pretreatment with a modified QuEChERS extraction and liquid chromatographic analysis with UV detection. The derivatized amino acids were separated on a HILIC column (NH2-HILIC 150 × 4.6 mm). The separation of 9 derivatized amino acids was successfully achieved with gradient elution. The derivatization process allowed high sensitivity for the amino acids while the rapid analysis in the HILIC mode within 17 min of elution time was obtained. Full separation and determination of the Dns-amino acids were achieved in relevant honey samples. Mean percentage recoveries of Dns-amino acids from the samples were 82.4% or higher with a good standard deviation of < 4%.

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References

  1. Alpert AJ (1990) Hydrophilic-interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. J Chromatogr A 499:177–196. https://doi.org/10.1016/S0021-9673(00)96972-3

    Article  CAS  Google Scholar 

  2. Gama MR, da Costa Silva RG, Collins CH, Bottoli CBG (2012) Hydrophilic interaction chromatography. Trends Anal Chem 37:48–60. https://doi.org/10.1016/j.trac.2012.03.009

    Article  CAS  Google Scholar 

  3. Cavazzini A, Catani M, Felinger A (2017) Liquid chromatography: fundamentals and ınstrumentation. 2nd edn.

  4. Marrubini G, Appelblad P, Maietta M, Papetti A (2018) Hydrophilic interaction chromatography in food matrices analysis: an updated review. Food Chem 257:53–66. https://doi.org/10.1016/j.foodchem.2018.03.008

    Article  CAS  PubMed  Google Scholar 

  5. Bernal J, Ares AM, Pól J, Wiedmer SK (2011) Hydrophilic interaction liquid chromatography in food analysis. J Chromatogr A 1218(42):7438–7452. https://doi.org/10.1016/j.chroma.2011.05.004

    Article  CAS  PubMed  Google Scholar 

  6. Dejaegher B, Vander Heyden Y (2010) HILIC methods in pharmaceutical analysis. J Sep Sci 33:697–715. https://doi.org/10.1002/jssc.200900742

    Article  CAS  Google Scholar 

  7. Donato P, Micalizzi G, Oteri M, Rigano F, Sciarrone D, Dugo P, Mondello L (2018) Comprehensive lipid profiling in the Mediterranean mussel (Mytilus galloprovincialis) using hyphenated and multidimensional chromatography techniques coupled to mass spectrometry detection. Anal Bioanal Chem 410:3297–3313. https://doi.org/10.1007/s00216-018-1045-3

    Article  CAS  PubMed  Google Scholar 

  8. Hemström P, Irgum K (2006) Hydrophilic interaction chromatography. J Sep Sci 29:1784–1821. https://doi.org/10.1002/jssc.200600199

    Article  CAS  PubMed  Google Scholar 

  9. Guo Y, Gaiki S (2011) Retention and selectivity of stationary phases for hydrophilic interaction chromatography. J Chromatogr A 1218(35):5920–5938. https://doi.org/10.1016/j.chroma.2011.06.052

    Article  CAS  PubMed  Google Scholar 

  10. Buszewski B, Noga S (2012) Hydrophilic interaction liquid chromatography (HILIC)—a powerful separation technique. Anal Bioanal Chem 402:231–247. https://doi.org/10.1007/s00216-011-5308-5

    Article  CAS  PubMed  Google Scholar 

  11. Aydoğan C, El Rassi Z (2016) Monolithic stationary phases with incorporated fumed silica nanoparticles. Part I. Polymethacrylate-based monolithic column with incorporated bare fumed silica nanoparticles for hydrophilic interaction liquid chromatography. J Chromatogr A 1445:55–61. https://doi.org/10.1016/j.chroma.2016.03.075

    Article  CAS  PubMed  Google Scholar 

  12. Aristoy MC, Toldra F. Amino acids in “Food Analysis by HPLC”. In: Nollet LML, Toldra F (eds) 3rd edn., CRC Press, USA

  13. Jaworska M, Stańczyk M, Wilk M, Kłaczkow G, Anuszewska E, Barzał J, Rzepecki P (2012) New approach for amino acid profiling in human plasma by selective fluorescence derivatization. Amino Acids 43:1653–1661. https://doi.org/10.1007/s00726-012-1243-9

    Article  CAS  PubMed  Google Scholar 

  14. Pereira V, Pontes M, Câmara JS, Marques JC (2008) Simultaneous analysis of free amino acids and biogenic amines in honey and wine samples using in loop orthophthalaldeyde derivatization procedure. J Chromatogr A 1189:435–443. https://doi.org/10.1016/j.chroma.2007.12.014

    Article  CAS  PubMed  Google Scholar 

  15. Rebane R, Herodes K (2010) A sensitive method for free amino acids analysis by liquid chromatography with ultraviolet and mass spectrometric detection using precolumn derivatization with diethyl ethoxymethylenemalonate: application to the honey analysis. Anal Chim Acta 672:79–84. https://doi.org/10.1016/j.aca.2010.04.014

    Article  CAS  PubMed  Google Scholar 

  16. Rebane R, Herodes K (2012) Comparison of three buffer solutions for amino acid derivatization and following analysis by liquid chromatography electrospray mass spectrometry. J Chromatogr A 1245:134–142. https://doi.org/10.1016/j.chroma.2012.05.039

    Article  CAS  PubMed  Google Scholar 

  17. Aydoǧan C, Andaç M, Bayram E, Say R, Denizli A (2012) Molecularly imprinted cryogel for l-glutamic acid separation. Biotechnol Progr 28(2):459–466. https://doi.org/10.1002/btpr.1517

    Article  CAS  Google Scholar 

  18. Aydoğan C (2018) Chiral separation and determination of amino acid enantiomers in fruit juice by open-tubular nano liquid chromatography. Chirality 30:1144–1149. https://doi.org/10.1002/chir.23006

    Article  CAS  PubMed  Google Scholar 

  19. Minocha R, Long S (2004) Simultaneous separation and quantitation of amino acids and polyamines of forest tree tissues and cell cultures within a single high-performance liquid chromatography run using dansyl derivatization. J Chromatogr A 1035:63–73. https://doi.org/10.1016/j.chroma.2004.02.026

    Article  CAS  PubMed  Google Scholar 

  20. Gros C, Labouesse B (1969) Study of the dansylation reaction of amino acids, peptides and proteins. Eur J Biochem 7:463–470. https://doi.org/10.1111/j.1432-1033.1969.tb19632

    Article  CAS  PubMed  Google Scholar 

  21. Tapuhi Y, Schmidt DE, Lindner W, Karger BL (1981) Dansylation of amino acids for high-performance liquid chromatography analysis. Anal Biochem 115:123–129

    Article  CAS  Google Scholar 

  22. Eurachem (1998) The fitness for purpose of analytical methods, a laboratory guide to method validation and related topics. Teddington

  23. Gómez-Alonso S, Hermosín-Gutiérrez I, García-Romero E (2007) Simultaneous HPLC analysis of biogenic amines, amino acids, and ammonium ion as aminoenone derivatives in wine and beer samples. J Agric Food Chem 55(3):608–613. https://doi.org/10.1021/jf062820m

    Article  CAS  PubMed  Google Scholar 

  24. McCalley DV (2010) Study of the selectivity, retention mechanisms and performance of alternative silica-based stationary phases for separation of ionised solutes in hydrophilic interaction chromatography. J Chromatogr A 1217(20):3408–3417. https://doi.org/10.1016/j.chroma.2010.03.011

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This study was partially funded by the Scientific Research Projects Coordination Unit of Bingöl University (Grant Number BAP-MMF.2018.00.002).

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Authors and Affiliations

  1. Central Laboratory, Bingöl University, Bingöl, Turkey

    Ertuğ Kırkan

  2. Department of Food Engineering, Bingöl University, Bingöl, Turkey

    Cemil Aydoğan

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  1. Ertuğ Kırkan
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  2. Cemil Aydoğan
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Correspondence to Cemil Aydoğan.

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Kırkan, E., Aydoğan, C. Free Amino Acid Analysis in Honey Samples by Hydrophilic Interaction Liquid Chromatography with UV Detection Using Precolumn Derivatization with Dansyl Chloride. Chromatographia 84, 127–133 (2021). https://doi.org/10.1007/s10337-020-03991-z

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