Analysis of Hydroxymethylfurfural in Honey-Flavored Syrups by HPLC

Ünal Savsar; Bilal Yılmaz

doi:10.17776/csj.1840832

Analysis of Hydroxymethylfurfural in Honey-Flavored Syrups by HPLC

Abstract

The aim of this study was to develop and validate a rapid, simple and reliable high-performance liquid chromatography (HPLC) method for the accurate determination of hydroxymethylfurfural (HMF) in commercially available honey-flavored syrups. Analyses were performed using a reverse-phase HPLC system equipped with a 250 × 4.6 mm, 5 µm C₁₈ column and a UV detector set at 284 nm. The mobile phase consisted of methanol and acetonitrile (10:90, v/v), delivered at a flow rate of 1.0 mL min^-1. Under these conditions, HMF was quantified in less than 6 minutes, demonstrating the method’s efficiency and suitability for routine analysis. Method validation followed International Council for Harmonisation guidelines. The calibration curve showed linearity across 0.10-100 µg mL^-1 with a correlation coefficient above 0.99. Precision values for both intra-day and inter-day measurements were below 2.37%, while accuracy showed relative error values under 0.87%. Recovery tests, conducted using the standard addition method on three honey-flavored syrups (GrinTuss, Biakaf, Bisolnatur), confirmed the method’s robustness. Overall, the developed HPLC procedure provides a straightforward, sensitive and highly reproducible approach for determining HMF levels, supporting quality control and safety assessment of honey-flavored pharmaceutical syrups. This ensures its applicability for routine monitoring in diverse quality control settings today

Keywords

Supporting Institution

The research reported in this study was funded by the Atatürk University Scientific Research Projects Coordination Unit under project number TYL-2024-14871.

Project Number

TYL-2024-14871

References

Baldwin, E. A., Bai, J., Plotto, A., Cameron, R., Luzio, G., Narciso, J., Manthey, J., Widmer, W., & Ford, B. L. (2012). Effect of extraction method on quality of orange juice: Hand-squeezed, commercial-fresh squeezed and processed. Journal of the Science of Food and Agriculture, 92(10), 2029-2042. https://doi.org/10.1002/jsfa.5587
Orfanakis, E., Vlasiadi, M., Philippidis, A., Zoumi, A., & Velegrakis, M. (2023). Rapid method for assessment of HMF levels in honey by absorption spectroscopy. Food Science and Engineering, 5(1), 26-33. https://doi.org/10.37256/fse.5120243640
Özbay, M., Arslan, F. N., & Görür, G. (2024). The use of HPLC-DAD method in determining honey quality: HMF analysis. Karamanoğlu Mehmetbey University Journal of Engineering and Natural Sciences, 6(2), 41-45. https://doi.org/10.55213/kmujens.1529151
Şahiner, İ., Göncü, A., Yıldırım Vardin, A., & Öğüt, S. (2025). Determination of HMF content and some physicochemical properties in granola bars with honey and molasses. Gıda, 50(5), 913-928. https://doi.org/10.15237/gida.GD25043
Belitz, H. D., & Grosch, W. (1999). Food chemistry (2nd ed.). Springer.
Arribas-Lorenzo, G., & Morales, F. J. (2010). Estimation of dietary intake of 5-hydroxymethylfurfural and related substances from coffee to Spanish population. Food and Chemical Toxicology, 48(2), 644-649. http://dx.doi.org/10.1016/j.fct.2009.11.046
Burdurlu, H. S., & Karadeniz, F. (2002). Maillard reaction products and their importance in food quality. Akademik Gıda, 2(2), 15–21.
Shapla, U. M., Solayman, M., Alam, N., Khalil, M. I., & Gan, S. H. (2018). 5‑Hydroxymethylfurfural (HMF) levels in honey and other food products: effects on bees and human health. Chemistry Central Hournal, 12(35), 1-18. https://doi.org/10.1186/s13065-018-0408-3

Tosi, E., Ciappini, M., Ré, E., & Lucero, H. (2002). Honey thermal treatment effects on hydroxymethylfurfural content. Food Chemistry, 77(1), 71-74. https://doi.org/10.1016/S0308-8146(01)00325-9
Uribarri, J., Del Castillo, M. D., De la Maza, M. P., Filip, R., Gugliucci, A., Luevano-Contreras, C., Macías-Cervantes, M. H., Markowicz Bastos, D. H., Medrano, A., Menini, T., Portero-Otin, M., Rojas, A., Rodrigues Sampaio, G., Wrobel, K., Wrobel, K., & Garay-Sevilla, M. E. (2015). Dietary advanced glycation end products and their role in health and disease. Advances in Nutrition, 6(4), 461-473. https://doi.org/10.3945/an.115.008433
Anonim. (2012). Türk Gıda Kodeksi Bal Tebliği (2012/58). Gıda Tarım ve Hayvancılık Bakanlığı, Ankara.
Rada-Mendoza M, Sanz, M. L., Olano, A., & Villamiel, M. (2004). Formation of hydroxymethylfurfural and furosine during the storage of jams and fruit-based infant foods. Food Chemistry, 85(4), 605-609. https://doi.org/10.1016/j.foodchem.2003.07.002
Kowalski, S., Lukasiewicz, M., Duda-Chodak, A., & Zięć, G. (2013). 5-Hydroxymethyl-2-furfural (HMF) - Heat-induced formation, occurrence in food and biotransformation - A review. Polish Journal of Food and Nutrition Sciences, 63(4), 207-225. https://doi.org/10.2478/v10222-012-0082-4
Zhang, R. L., Zhang, M. M., Pu, Y. F., & Zhu, L. X. (2022). Evolution of nonenzymatic browning during the simulated Msalais‑production process in models of grape juice. Food Science & Nutrition, 10(7), 2132-2140. https://doi.org/10.1002/fsn3.2829
Muslu, A., Üçüncü, O., & Baltacı, C. (2024). Production of 5-hydroxymethylfurfural from non-edible lignocellulosic food wastes. BioResources, 19(3), 5752-5763. https://doi.org/10.15376/biores.19.3.5752-5763
Adu, J. K., Orman, E., Amengor, C. D. K., Peprah, P., Danso, N., & Azumah, J. A. (2022). Estimation of 5-hydroxymethylfurfural levels in carbonated soft drinks and milk products on the markets in the Kumasi Metropolis of Ghana. Scientific African, 17, e01346. https://doi.org/10.1016/j.sciaf.2022.e01346
Janzowski, C., Glaab, V., Samimi, E., Schlatter, J., & Eisenbrand, G. (2000). 5-HMF: Mutagenicity, DNA damage, and reactivity with glutathione. Food and Chemical Toxicology, 38(9), 801-809. https://doi.org/10.1016/s0278-6915(00)00070-3
Özdal, H. R., Yıldız, B., & Arkun, G. (2018). Hydroxymethylfurfural (HMF) formation in milk and dairy products. International Journal of Food Engineering Research (IJFER), 4(1), 13-23.
Meng, D., Zhang, F., Jia, W., Jiao, J., & Zhang, Y. (2024). Understanding the health implications of furan and its derivatives in thermally processed foods: Occurrence, toxicology, and mixture analysis. Current Opinion in Food Science, 57, 101151. https://doi.org/10.1016/j.cofs.2024.101151
Babacanoğlu, C., Tuğbagül Çal Doğan, T., Aydın Dilsiz, S., Bucurgat, M., & Ündeğer Bucurgat, Ü. (2025). In vitro evaluation of the possible genotoxic effects of acrylamide and 5-hydroxymethylfurfural by single cell gel electrophoresis (Comet) assay. European Food Research and Technology, 251, 2779-2791. https://doi.org/10.1007/s00217-025-04793-z
Rufian-Henares, J. A., & De La Cueva, S. P. (2007). Assessment of HMF intake from Spanish food market. Food Research International, 40(7), 807–813. https://doi.org/10.1080/02652030802163406
Dybing, E., O’Brien, J., Renwick, A. G., & Sanner, T. (2008). Risk assessment of dietary exposures to compounds that are genotoxic and carcinogenic-An overview. Toxicology Letters, 180(2), 110-117. https://doi.org/10.1016/j.toxlet.2008.05.007
Aykas, D. P. (2023). Determination of possible adulteration and quality assessment in commercial honey. Foods, 12(3), 523. https://doi.org/10.3390/foods12030523
Gün, R., & Karaoğlu, M. M. (2024). Detection of honey adulteration by characterization of physicochemical properties of honey with glucose-fructose and maltose syrups. European Food Research and Technology, 250, 2255-2272. https://doi.org/10.1007/s00217-024-04535-7
Korkmaz, S. D. (2023). Determination of HMF value and diastase activities in strained honeys sold in markets. Black Sea Journal of Engineering and Science, 6(4), 522-526. https://doi.org/10.34248/bsengineering.1349150
The United States Pharmacopoeia and The National Formulary (30th & 25th ed.). (2007). United States Pharmacopeial Convention.
ICH Topic Q2B: Validation of Analytical Procedures. (1996). The European Agency for the Evaluation of Medicinal Products.

Details

Primary Language

English

Subjects

Separation Science , Analytical Chemistry (Other)

Journal Section

Research Article

Authors

Ünal Savsar
0009-0008-1972-3360
Türkiye

Bilal Yılmaz ^*
0000-0002-8574-7570
Türkiye

Publication Date

April 29, 2026

Submission Date

December 15, 2025

Acceptance Date

April 12, 2026

Published in Issue

Year 2026 Volume: 47 Number: 2

DOI

https://doi.org/10.17776/csj.1840832

IZ

https://izlik.org/JA63TH24WH

Cite

RIS / Bibtex

APA

Savsar, Ü., & Yılmaz, B. (2026). Analysis of Hydroxymethylfurfural in Honey-Flavored Syrups by HPLC. Cumhuriyet Science Journal, 47(2), 343-349. https://doi.org/10.17776/csj.1840832