Abstract
Honey is a complex mixture of carbohydrates, in which the monosaccharides glucose and fructose are the most abundant compounds. Currently, more than 20 oligosaccharides have been identified in different varieties of honey normally at quite low concentration. A method was developed and validated using high-performance anion-exchange chromatography coupled to a mass spectrometry detector to investigate the composition of carbohydrates in honey samples. The method was tested for linearity range, trueness, instrumental and method detection and quantification limits, repeatability, and reproducibility. It was applied to determine seven monosaccharides, eight disaccharides, four trisaccharides, and one tetrasaccharide in various honey samples. The present work describes the composition of sugars in unifloral, multifloral, and some honeydew honey, which were produced and collected by beekeepers in the Trentino Alto-Adige region. Statistical techniques have been used to establish a relationship based on levels of carbohydrates among different Italian honey. The results emphasize that mono- and oligosaccharide profiles can be useful to discriminate different honeys according to their floral characteristics and inter-annual variability.
Similar content being viewed by others
References
Da Costa Leite JM, Trugo LC, Costa LSM, Quinteiro LMC, Barth OM, Dutra VML, et al. Determination of oligosaccharides in Brazilian honeys of different botanical origin. Food Chem. 2000;70:93–8. https://doi.org/10.1016/S0956-7135(99)00115-2.
Pita-Calvo C, Guerra-Rodríguez ME, Vázquez M. Analytical methods used in the quality control of honey. J Agric Food Chem. 2017;65:690–703. https://doi.org/10.1021/acs.jafc.6b04776.
Thrasyvoulou A, Tananaki C, Goras G, Karazafiris E, Dimou M, Liolios V, et al. Legislación de criterios y normas de miel. J Apic Res. 2018;57:88–96.
Al-Qassemi R, Robinson RK. Some special nutritional properties of honey – a brief review. Nutr Food Sci. 2003;33:254–60. https://doi.org/10.1108/00346650310507073.
Buba F, Gidado A, Shugaba A. Analysis of biochemical composition of honey samples from north-East Nigeria. Biochem Anal Biochem. 2013;2. https://doi.org/10.4172/2161-1009.1000139.
Arias VC, Castells RC, Malacalza N, Lupano CE, Castells CB. Determination of oligosaccharide patterns in honey by solid-phase extraction and high-performance liquid chromatography. Chromatographia. 2003;58:797–801. https://doi.org/10.1365/s10337-003-0115-6.
De La Fuente E, Ruiz-Matute AI, Valencia-Barrera RM, Sanz J, Martínez Castro I. Carbohydrate composition of Spanish unifloral honeys. Food Chem. 2011;129:1483–9. https://doi.org/10.1016/j.foodchem.2011.05.121.
Ruiz-Matute AI, Brokl M, Soria AC, Sanz ML, Martínez-Castro I. Gas chromatographic-mass spectrometric characterisation of tri- and tetrasaccharides in honey. Food Chem. 2010;120:637–42. https://doi.org/10.1016/j.foodchem.2009.10.050.
Doner LW. The sugars of honey—a review. J Sci Food Agric. 1977;28:443–56. https://doi.org/10.1002/jsfa.2740280508.
Escuredo O, Dobre I, Fernández-González M, Seijo MC. Contribution of botanical origin and sugar composition of honeys on the crystallization phenomenon. Food Chem. 2014;149:84–90. https://doi.org/10.1016/j.foodchem.2013.10.097.
Ouchemoukh S, Schweitzer P, Bachir Bey M, Djoudad-Kadji H, Louaileche H. HPLC sugar profiles of Algerian honeys. Food Chem. 2010;121:561–8. https://doi.org/10.1016/j.foodchem.2009.12.047.
Zhou Y, Xu DS, Liu L, Qiu FR, Chen JL, Xu GL. A LC-MS/MS method for the determination of stachyose in rat plasma and its application to a pharmacokinetic study. J Pharm Biomed Anal. 2016;123:24–30. https://doi.org/10.1016/j.jpba.2015.11.041.
Al Somal N, Coley KE, Molan PC, Hancock BM. Susceptibility of helicobacter pylori to the antibacterial activity of manuka honey. J R Soc Med. 1994;87:9–12. https://doi.org/10.1177/014107689408700106.
Montilla A, Van De Lagemaat J, Olano A, Del Castillo MD. Determination of oligosaccharides by conventional high-resolution gas chromatography. Chromatographia. 2006;63:453–8. https://doi.org/10.1365/s10337-006-0770-5.
Trugo LC, Farah A, Cabral L. Oligosaccharide distribution in Brazilian soya bean cultivars. Food Chem. 1995;52:385–7. https://doi.org/10.1016/0308-8146(95)93286-Z.
Zakharova AM, Grinshtein IL, Kartsova LA. Determination of carbohydrates and sweeteners in foods and biologically active additives by high-performance liquid chromatography. J Anal Chem. 2013;68:1081–4. https://doi.org/10.1134/S1061934813100122.
Goodall I, Dennis MJ, Parker I, Sharman M. Contribution of high-performance liquid chromatographic analysis of carbohydrates to authenticity testing of honey. J Chromatogr A. 1995;706:353–9. https://doi.org/10.1016/0021-9673(94)01074-O.
Martínez Montera C, Rodríguez Dodero MC, Guillén Sánchez DA, Barroso CG. Analysis of Low molecular weight carbohydrates in food and beverages: a review. Chromatographia. 2004;59:15–30. https://doi.org/10.1365/s10337-003-0134-3.
Swallow KW, Low NH. Analysis and quantitation of the carbohydrates in honey using high-performance liquid chromatography. J Agric Food Chem. 1990;38:1828–32. https://doi.org/10.1021/jf00099a009.
Sanz ML, Polemis N, Morales V, Corzo N, Drakoularakou A, Gibson GR, et al. In vitro investigation into the potential prebiotic activity of honey oligosaccharides. J Agric Food Chem. 2005;53:2914–21. https://doi.org/10.1021/jf0500684.
Anjos O, Campos MG, Ruiz PC, Antunes P. Application of FTIR-ATR spectroscopy to the quantification of sugar in honey. Food Chem. 2015;169:218–23. https://doi.org/10.1016/j.foodchem.2014.07.138.
Jan Mei S, Mohd Nordin MS, Norrakiah AS. Fructooligosaccharides in honey and effects of honey on growth of Bifidobacterium longum BB 536. Int Food Res J. 2010;17:557–61.
Mateo R, Bosch-Reig F. Sugar profiles of Spanish unifloral honeys. Food Chem. 1997;60:33–41. https://doi.org/10.1016/S0308-8146(96)00297-X.
Cotte JF, Casabianca H, Chardon S, Lheritier J, Grenier-Loustalot MF. Application of carbohydrate analysis to verify honey authenticity. J Chromatogr A. 2003;1021:145–55. https://doi.org/10.1016/j.chroma.2003.09.005.
Cano CB, Felsner ML, Bruns RE, Matos JR, Almeida-Muradi LB. Optimization of mobile phase for separation of carbohydrates in honey by high performance liquid chromatography using a mixture design. J Braz Chem Soc. 2006;17:588–93. https://doi.org/10.1590/S0103-50532006000300024.
Weston RJ, Brocklebank LK. The oligosaccharide composition of some New Zealand honeys. Food Chem. 1999;64:33–7. https://doi.org/10.1016/S0308-8146(98)00099-5.
Ruiz-Matute AI, Sanz ML, Martinez-Castro I. Use of gas chromatography-mass spectrometry for identification of a new disaccharide in honey. J Chromatogr A. 2007;1157:480–3. https://doi.org/10.1016/j.chroma.2007.05.056.
Terrab A, Vega-Pérez JM, Díez MJ, Heredia FJ. Characterisation of northwest Moroccan honeys by gas chromatographic-mass spectrometric analysis of their sugar components. J Sci Food Agric. 2002;82:179–85. https://doi.org/10.1002/jsfa.1011.
Bogdanov S. Harmonised methods of the International Honey Commission. Bee Prod Sci. 2009;1–62. https://doi.org/10.1007/s13398-014-0173-7.2.
Morales V, Sanz ML, Olano A, Corzo N. Rapid separation on activated charcoal of high oligosaccharides in honey. Chromatographia. 2006;64:233–8. https://doi.org/10.1365/s10337-006-0842-6.
Bruggink C, Maurer R, Herrmann H, Cavalli S, Hoefler F. Analysis of carbohydrates by anion exchange chromatography and mass spectrometry. J Chromatogr A. 2005;1085:104–9 https://doi.org/10.1016/j.chroma.2005.03.108.
Corradini C, Cavazza A, Bignardi C. High-performance anion-exchange chromatography coupled with pulsed electrochemical detection as a powerful tool to evaluate carbohydrates of food interest: principles and applications. Int J Carbohydr Chem. 2012;2012:1–13. https://doi.org/10.1155/2012/487564.
Gambaro A, Barbaro E, Zangrando R, Barbante C. Simultaneous quantification of microcystins and nodularin in aerosol samples using high-performance liquid chromatography/negative electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom. 2012;26:1497–506. https://doi.org/10.1002/rcm.6246.
Barbaro E, Kirchgeorg T, Zangrando R, Vecchiato M, Piazza R, Barbante C, et al. Sugars in Antarctic aerosol. Atmos Environ. 2015;118:135–44. https://doi.org/10.1016/j.atmosenv.2015.07.047.
Morales V, Corzo N, Sanz ML. HPAEC-PAD oligosaccharide analysis to detect adulterations of honey with sugar syrups. Food Chem. 2008;107:922–8. https://doi.org/10.1016/j.foodchem.2007.08.050.
Rybak-Chmielewska H, Szczesna T. Determination of saccharides in multifloral honey by means of HPLC. J Apic Sci. 2003;47:93.
GE Healthcare. Application Note AA 28-9372-07. Column efficiency testing. Uppsala: GE Healthcare; 2010;1–5.
Samanidou VF. Basic LC method development and optimization. Anal Sep Sci. 2015;25–42. https://doi.org/10.1002/9783527678129.assep002.
Ardrey RE. Liquid chromatography–mass spectrometry: an introduction. Chichester: Wiley; 2003.
Tezcan F, Kolayli S, Ulusoy HSE, Erim FB. Evaluation of organic acid, saccharide composition and antioxidant properties of some authentic Turkish honeys. J Food Nutr Res. 2011;50:33–40.
Puscas A, Hosu A, Cimpoiu C. Application of a newly developed and validated high-performance thin-layer chromatographic method to control honey adulteration. J Chromatogr A. 2013;1272:132–5. https://doi.org/10.1016/j.chroma.2012.11.064.
Tůma P, Málková K, Samcová E, Štulík K. Rapid monitoring of mono- and disaccharides in drinks, foodstuffs and foodstuff additives by capillary electrophoresis with contactless conductivity detection. Anal Chim Acta. 2011;698:1–5. https://doi.org/10.1016/j.aca.2011.04.055.
Bliesner DM. Validating chromatographic methods a practical guide. Hoboken: Wiley; 2006.
Val A, Huidobro JF, Sánchez MP, Muniategui S, Fernández-Muiño MA, Sancho MT. Enzymatic determination of galactose and lactose in honey. J Agric Food Chem. 1998;46:1381–5. https://doi.org/10.1021/jf970483z.
Anjos O, Santos AJA, Paixão V, Estevinho LM. Physicochemical characterization of Lavandula spp. honey with FT-Raman spectroscopy. Talanta. 2018;178:43–8. https://doi.org/10.1016/j.talanta.2017.08.099.
Gómez Bárez JA, Garcia-Villanova RJ, Elvira Garcia S, González Paramás AM. Optimization of the capillary gas chromatographic analysis of mono- and oligosaccharides in honeys. Chromatographia. 1999;50:461–9. https://doi.org/10.1007/BF02490743.
Azeredo LDC, Azeredo MAA, De Souza SR, Dutra VML. Protein contents and physicochemical properties in honey samples of Apis mellifera of different floral origins. Food Chem. 2003;80:249–54. https://doi.org/10.1016/S0308-8146(02)00261-3.
Acknowledgments
This work was co-founded by Edmund Mach Foundation under the Research Centre International Program for PhD projects. The authors are grateful to Franco Abballe and Diego Visentin (Thermo Fischer Scientific) for technical instrumental support and their help and cooperation during the activities. We would like to thank the apiarist associations located in the Trentino Alto-Adige region (Italy) and the local beekeeper farms for providing the honey samples. The authors thank Elga LabWater, High Wycombe UK for supplying the pure water systems used in this study. Grateful acknowledgment is also extended to Dr. Massimiliano Vardè (CNR-ISP) for his useful support on organizing the references over the entire manuscript. The authors are grateful to Dr. Nicole Emilia Byrne for her intense English revision of the entire manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interests
The authors declare that they have no conflict of interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(PDF 174 kb)
Rights and permissions
About this article
Cite this article
Tedesco, R., Barbaro, E., Zangrando, R. et al. Carbohydrate determination in honey samples by ion chromatography–mass spectrometry (HPAEC-MS). Anal Bioanal Chem 412, 5217–5227 (2020). https://doi.org/10.1007/s00216-020-02732-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-020-02732-3