Analytical and Bioanalytical Chemistry

, Volume 408, Issue 7, pp 1871–1878 | Cite as

Analysis of saccharides in beverages by HPLC with direct UV detection

  • Thomas Schmid
  • Benedikt Baumann
  • Markus Himmelsbach
  • Christian W. Klampfl
  • Wolfgang Buchberger
Research Paper


The present study demonstrates the suitability of direct UV detection for saccharide analysis in HPLC. Under highly alkaline conditions, the non-UV absorbing saccharides are converted by a photo-initiated chemical reaction in the detection cell into malonenolate, which can be detected at 266 nm. A straightforward method for such direct UV detection of saccharides after their separation by anion-exchange chromatography was developed and successfully applied to several beverage samples. Investigation and optimization of the influencing factors using design of experiment resulted in a baseline separation of glucose, fructose, and sucrose within 6 min and LOD values below 0.2 mg L−1. In addition, a fast, simple and cost-effective flow injection method was developed to estimate the total saccharide concentration. The results of this method applied to beverage samples are in good agreement with the chromatographic method as well as to the saccharide concentration stated by the manufacturer. Finally, a comparison of different commercially available UV detectors and detector cells revealed that sensitive detection requires the use of recently introduced flow cells with extended path length.

Graphical Abstract

Online direct UV detection of saccharides in HPLC, which is possible due to a photo-initiated conversion of the saccharides into malonenolate


Anion-exchange chromatography Direct UV detection HPLC Saccharide 



The authors thank Andreas Kitzler from Agilent Technologies for technical support regarding HPLC instrumentation.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Martínez Montero C, Rodríguez Dodero M, Guillén Sánchez D, Barroso C. Analysis of low molecular weight carbohydrates in food and beverages: a review. Chromatographia. 2004;59:15–30.Google Scholar
  2. 2.
    Galant AL, Kaufman RC, Wilson JD. Glucose: detection and analysis. Food Chem. 2015;188:149–60.CrossRefGoogle Scholar
  3. 3.
    Ruiz-Matute A, Hernández-Hernández O, Rodríguez-Sánchez S, Sanz M, Martínez-Castro I. Derivatization of carbohydrates for GC and GC–MS analyses. J Chromatogr B. 2011;879:1226–40.CrossRefGoogle Scholar
  4. 4.
    Harvey DJ. Derivatization of carbohydrates for analysis by chromatography; electrophoresis and mass spectrometry. J Chromatogr B. 2011;879:1196–225.CrossRefGoogle Scholar
  5. 5.
    El Rassi Z. Carbohydrate analysis by modern chromatography and electrophoresis. 1st ed. Amsterdam: Elsevier; 2002.Google Scholar
  6. 6.
    Jandera P. Stationary and mobile phases in hydrophilic interaction chromatography: a review. Anal Chim Acta. 2011;692:1–25.CrossRefGoogle Scholar
  7. 7.
    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. doi: 10.1155/2012/487564 .Google Scholar
  8. 8.
    Rohrer JS, Basumallick L, Hurum D. High-performance anion-exchange chromatography with pulsed amperometric detection for carbohydrate analysis of glycoproteins. Biochem Mosc. 2013;78:697–709.CrossRefGoogle Scholar
  9. 9.
    García-Cañas V, Simó C, Castro-Puyana M, Cifuentes A. Recent advances in the application of capillary electromigration methods for food analysis and foodomics. Electrophoresis. 2014;35:147–69.CrossRefGoogle Scholar
  10. 10.
    Rovio S, Yli-Kauhaluoma J, Sirén H. Determination of neutral carbohydrates by CZE with direct UV detection. Electrophoresis. 2007;28:3129–35.CrossRefGoogle Scholar
  11. 11.
    Sarazin C, Delaunay N, Costanza C, Eudes V, Mallet J, Gareil P. New avenue for Mid-UV-range detection of underivatized carbohydrates and amino acids in capillary electrophoresis. Anal Chem. 2011;83:7381–7.CrossRefGoogle Scholar
  12. 12.
    Oliver JD, Gaborieau M, Hilder EF, Castignolles P. Simple and robust determination of monosaccharides in plant fibers in complex mixtures by capillary electrophoresis and high performance liquid chromatography. J Chromatogr A. 2013;1291:179–86.CrossRefGoogle Scholar
  13. 13.
    Oliver JD, Rosser AA, Fellows CM, Guillaneuf Y, Clement J, Gaborieau M, et al. Understanding and improving direct UV detection of monosaccharides and disaccharides in free solution capillary electrophoresis. Anal Chim Acta. 2014;809:183–93.CrossRefGoogle Scholar
  14. 14.
    Schmid T, Himmelsbach M, Oliver JD, Gaborieau M, Castignolles P, Buchberger W. Investigation of photochemical reactions of saccharides during direct ultraviolet absorbance detection in capillary electrophoresis. J Chromatogr A. 2015;1388:259–66.CrossRefGoogle Scholar
  15. 15.
    Schmid T, Himmelsbach M, Buchberger WW. Investigation of photochemical reaction products of glucose formed during direct UV detection in capillary electrophoresis. Electrophoresis. 2015. doi: 10.1002/elps.201500283 .Google Scholar
  16. 16.
    Varandas S, Teixeira MJ, Marques JC, Aguiar A, Alves A, Bastos MM. Glucose and fructose levels on grape skin: interference in Lobesia botrana behaviour. Anal Chim Acta. 2004;513:351–5.CrossRefGoogle Scholar
  17. 17.
    Aristoy MC, Orlando L, Navarro JL, Sendra JM, Izquierdo L. Characterization of Spanish orange juice for variables used in purity control. J Agric Food Chem. 1989;37:596–600.CrossRefGoogle Scholar
  18. 18.
    Villamiel M, Martínez-Castro I, Olano A, Corzo N. Quantitative determination of carbohydrates in orange juice by gas chromatography. Z Lebensm Unters Forsch. 1998;206:48–51.CrossRefGoogle Scholar
  19. 19.
    Fuleki T, Pelayo E, Palabay RB. Sugar composition of varietal juices produced from fresh and stored apples. J Agric Food Chem. 1994;42:1266–75.CrossRefGoogle Scholar
  20. 20.
    Karadeniz F, Ekşi A. Sugar composition of apple juices. Eur Food Res Technol. 2002;215:145–8.CrossRefGoogle Scholar
  21. 21.
    Vochyánová B, Opekar F, Tůma P, Štulík K. Rapid determinations of saccharides in high-energy drinks by short-capillary electrophoresis with contactless conductivity detection. Anal Bioanal Chem. 2012;404:1549–54.CrossRefGoogle Scholar
  22. 22.
    Takayanagi T, Yokotsuka K. Relationship between sucrose accumulation and sucrose-metabolizing enzymes in developing grapes. Am J Enol Vitic. 1997;4:403–7.Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Thomas Schmid
    • 1
  • Benedikt Baumann
    • 1
  • Markus Himmelsbach
    • 1
  • Christian W. Klampfl
    • 1
  • Wolfgang Buchberger
    • 1
  1. 1.Institute of Analytical ChemistryJohannes Kepler University LinzLinzAustria

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