Single Laboratory Validation of Four Methods for Quantification of HMF in Honey

Abstract

Hydroxymethylfurfural or 5-(hydroxymethyl)-2-furaldehyde (HMF) has been absent or found in honey naturally at very low amount. It is formed in honey mainly by heating process and improper storage conditions. HMF has been reported to have cytotoxic, carcinogenic, and mutagenic effects and thus regulatory agencies in many countries impose restrictions on its maximum levels in honey. Validated methods have been required for effective and specific detection and quantification of HMF in food samples. In this study, a single laboratory validation study was conducted on four quantification methods: direct spectral analysis, High Performance Liquid Chromatography (HPLC) analysis, Seliwanoff and Winkler methods. All methods showed linearity with the lowest R2 value of 0.992. Two method performance parameters, accuracy, and precision were satisfied by each four methods with recovery values at 98.2%, 100.2%, 102.5% and 103.3% and RSDr (relative standard deviation) % values at 6.97%, 6.19%, 2.87% and 0.90% for spectral analysis, Seliwanoff, HPLC and Winkler methods, respectively. Based on the measurement uncertainties of four quantification methods, honey samples spiked with HMF at the final concentration of 0.004mg/0.1g were reported as 0.004 mg/0.1g ± 0.00025 mg/0.1g by spectral analysis, 0.0036 mg/0.1g ± 0.000691 mg/0.1g by Seliwanoff method, 0.004 mg/0.1g ± 0.00045 mg/0.1g by HPLC and 0.0039 mg/0.1g ± 0.00022 mg/0.1g by Winkler methods (k=2, confidence level of 95%). The validated methods can quantify HMF in honey with a target concentration of 0.004 mg/0.1g, specifically and accurately.

Keywords

• HMF
• honey
• method validation
• Seliwanoff
• uncertainty

References

1. D. B. Markowicz, É. Monaro, É., Siguemoto, M. Séfora, Maillard reaction products in processed foods: Pros and cons, in: B. Valdez (Ed.), Food Industrial Processes-Methods and Equipment, London: INTECH Open Access Publisher, Rijeka, 2012, Ch. 15, pp. 281–300.
2. N. İçli, Evaluation of HMF levels in unbranded flower honeys in terms of food safety, Ankara Üniversitesi Veteriner Fakültesi Dergisi 69 (4) (2022) 431–436.
3. U. M. Shapla, M. Solayman, N. Alam, M. I. Khalil, S. H. Gan, 5-Hydroxymethylfurfural (HMF) levels in honey and other food products: effects on bees and human health, Chemistry Central Journal 12 (2018) Article Number 35 18 pages.
4. C. Janzowski, V. Glaab, E. Samımı, J. Schlatter, G. Eisebbrand, 5-Hydroxymethylfurfural: Assessment of mutagenicity, DNA-damaging potential and reactivity towards cellular glutathione, Food and Chemical Toxicology 38 (9) (2000) 801–809.
5. E. Capuano, V. Fogliano, Acrylamide and 5-hydroxymethylfurfural (HMF): A review on metabolism, toxicity, occurrence in food and mitigation strategies, LWT - Food Science and Technology 44 (4) (2011) 793–810.
6. M. N. Islam, M. I. Khalil, M. A. Islam, S. H. Gan, Toxic compounds in honey, Journal of Applied Toxicology, 34 (7) (2014) 733–742.
7. A. Apriceno, R. Bucci, A. M. Girelli, F. Marini, L. Quattrocchi, 5-Hydroxymethyl furfural determination in Italian honeys by a fast near infrared spectroscopy, Microchemical Journal 143 (2018) 140–144.
8. M. C. Archer, W. R. Bruce, C. C. Chan, D. E. Corpet, A. Medline, L. Roncucci, D. Stamp, X. M. Zhang, Aberrant crypt foci and microadenoma as markers for colon cancer, Environmental Health Perspectives 98 (1992) 195–197.

9. W. R. Bruce, M. C. Archer, D. E. Corpet, A. Medline, S. Minkin, D. Stamp, Y. Yin, X.-M. Zhang, Diet, aberrant crypt foci and colorectal cancer, Mutation Research 290 (1) (1993) 111–118.
10. X. M. Zhang, C. C. Chan, D. Stamp, S. Minkin, M. C. Archer, W. R. Bruce, Initiation and promotion of colonic aberrant crypt foci in rats by 5-hydroxymethyl-2-furaldehyde in thermolyzed sucrose, Carcinogenesis 14 (4) (1993) 773–775.
11. Y. C. Lee, M. Shlyankevich, H. K. Jeong, J. S. Douglas, Y. J. Surh, Bioactivation of 5-hydroxymethyl-2-furaldehyde to an electrophilic and mutagenicallylic sulfuric acid ester, Biochemical and Biophysical Research Communications 209 (3) (1995) 996–1002.
12. B. H. Monien, W. Engst, G. Barknowitz, A. Seidel, H. Glatt, Mutagenicity of 5-hydroxymethylfurfural in V79 cells expressing human SULT1A1: identification and mass spectrometric quantification of DNA adducts formed, Chemical Research in Toxicology 25 (7) (2012) 1484–92.
13. Joint FAO/WHO Codex Alimentarius Commission, Codex Alimentarius: Report of the Seventh Session of the Codex Committee on Sugars ALINORM 01/25 (2000), https://www.fao.org/input/download/report/277/Al01_25e.pdf Accessed 28 Feb 2024
14. The Council of the European Union, Council Directive 2001/110/EC relating to honey (2001), https://www.fao.org/faolex/results/details/en/c/LEX-FAOC037441 Accessed 28 Feb 2024
15. Turkish Food Codex, Turkish Food Codex Honey Statement 2020/7 (2020), https://www.resmigazete.gov.tr/eskiler/2020/04/20200422-13.htm Accessed 28 Feb 2024
16. F. J. Morales, C. Romero, S. Jiménez-Pérez, Chromatographic determination of bound hydroxymethylfurfural as an index of milk protein glycosylation, Journal of Agricultural and Food Chemistry 45 (5) (1997) 1570–1573.
17. G. Vanhoenacker, A. D. Villiers, K. Lazou, D. D. Keukeleire, P. Sandra, Comparison of high-performance liquid chromatography - Mass spectroscopy and capillary electrophoresis - Mass spectroscopy for the analysis of phenolic compounds in diethyl ether extracts of red wines, Chromatographia 54 (2001) 309–315.
18. B. M. Wagner, F. Donnarumma, R. Wintersteiger, W. Windischhofer, H. J. Leis, Simultaneous quantitative determination of alpha-ketoglutaric acid and 5-hydroxymethylfurfural in human plasma by gas chromatography-mass spectrometry, Analytical and Bioanalytical Chemistry 396 (7) (2010) 2629–2637.
19. D. Jöbstl, T. Husoy, J. Alexander, T. Bjellaas, E. Leitner, M. Murkovic, Analysis of 5-hydroxymethyl-2-furoic acid (HMFA) the main metabolite of alimentary 5-hydroxymethyl-2-furfural (HMF) with HPLC and GC in urine, Food Chemistry 123 (3) (2010) 814–818.
20. S.-M. Lin, J.-Y. Wu, C. Su, S. Ferng, C.-Y. Lo, R. Y.-Y. Chiou, Identification and mode of action of 5-hydroxymethyl-2-furfural (5-HMF) and 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (MTCA) as potent xanthine oxidase inhibitors in vinegars, Journal of Agricultural and Food Chemistry 60 (39) (2012) 9856–9862.
21. Y. Huang, C. Li, H. Hu, Y. Wang, M. Shen, S. Nie, J. Chen, M. Zeng, M. Xie, Simultaneous Determination of Acrylamide and 5-Hydroxymethylfurfural in Heat-Processed Foods Employing Enhanced Matrix Removal-Lipid as a New Dispersive Solid-Phase Extraction Sorbent Followed by Liquid Chromatography-Tandem Mass Spectrometry. Journal of Agricultural and Food Chemistry 67 (17) (2019) 5017–5025.
22. S. Bogdanov, C. Lüllmann, P. Martin, W. von der Ohe, H. Russmann, G. Vorwohl, L. P. Oddo, A.-G. Sabatini, G. L. Marcazzan, R. Piro, C. Flamini, M. Morlot, J. Lhéritier, R. Borneck, P. Marioleas, A. Tsigouri, J. Kerkvliet, A. Ortiz, T. Ivanov, B. D’Arcy, B. Mossel, P. Vit, Honey Quality and International Regulatory Standards: Review by the International Honey Commission, Bee World 80 (2) (1999) 61–69.
23. H. J. Jeuring, F. J. E. M. Kuppers, High performance liquid chromatography of furfural and hydroxymethylfurfural in spirits and honey, Journal of the Association of Official Analytical Chemists 63 (6) (1980) 1215–1218.
24. O. Winkler, Beitrag zum Nachweis und zur bestimmung von oxymethylfurfural in Honig und Kunsthonig, Zeitschrift fur Lebensmittel Untersuchung und Forshung 102 (3) (1955) 161–167.
25. J. White, Spectrophotometric method for hydroxymethylfurfural in honey, Journal of the Association of Official Analytical Chemists 62 (3) (1979) 509–514.
26. S. S. Virgen, S. G. Ceballos-Magana, V. D. C. Salvatierre-Stamp, M. T. Sumaya-Martinez, F. J. Martinez-Mertinez, R. Muniz-Valencia, HPLC-DAD method development and validation for the quantification of hydroxymethylfurfural in corn chips by means of response surface optimization, Food Additives & Contaminants: Part A 34 (12) (2017) 2101–2110.
27. M. Zappalà, B. Fallico, E. Arena, A. Verzera, Methods for the determination of HMF in honey: A comparison, Food Control 16 (3) (2005) 273–277.
28. A. Elmastaş, F. Aydın, A. Umaz, E. Kılınç, Y. Arslan, I. Aydın, Determination of hydroxymethylfurfural in Turkish honeys, pekmez (grape molasses), and jam samples by high-performance liquid chromatography with diode array detection, Journal of Food Processing and Preservation 46 (4) (2022) e16462 7 pages.
29. International Organization for Standardization, Accuracy (trueness and precision) of measurement methods and results Part 2 ISO 5725-2 (1994), https://www.iso.org/standard/36202.html Accessed 28 Feb 2024
30. E. Yıldız, Determination of hydroxymethyl furfural in honey, Master’s Thesis Karamanoğlu Mehmetbey University (2019) Karaman.
31. B. F. M. Kuster, 5-Hydroxymethylfurfural (HMF). A review focussing on its manufacture, Starch Stärke 42 (8) (1990) 314–321.
32. B. Fallico, M. Zappalà, E. Arena, A. Verzera, Effects of conditioning on HMF content in unifloral honeys, Food Chemistry 85 (2) (2004) 305–313.
33. V. Gökmen, Ö. Ç. Açar, H. Köksel, J. Acar, Effects of dough formula and baking conditions on acrylamide and hydroxymethylfurfural formation in cookies, Food Chemistry 104 (3) (2007) 1136–1142.
34. H. S. Lee, S. Nagy, Relative reactivities of sugars in the formation of 5-hydroxymethylfurfural in sugar-catalyst model systems, Journal of Food Processing and Preservation 14 (3) (1990) 171–178.
35. Y. Román-Leshkov, J. N. Chheda, J. A. Dumesic, Phase modifiers promote efficient production of hydroxymethylfurfural from fructose, Science 312 (5782) (2006) 1933–1937.
36. J. H. Turner, P. A. Rebers, P. L. Barrick, R. H. Cotton, Determination of 5‐(hydroxymethyl)‐2‐furaldehyde and related compounds, Analytical Chemistry 26 (5) (1954) 898–901.
37. N. Hudz, D. Leontiev, P. P. Wieczorek, Spectral characteristics of 5-hydroxymethylfurfural as a related substance in medicinal products containing glucose, Pharmacia 66 (3) (2019) 121–125.
38. S. Bogdanov, Harmonised methods of the European Honey Commission (2009), http://www.ihc-platform.net/ihcmethods2009.pdf Accessed 28 Feb 2024
39. C. Truzzi, A. Annibaldi, S. Illuminati, C. Finale, M. Rossetti, G. Scarponi, Determination of very low levels of 5- (hydroxymethyl)-2-furaldehyde (HMF) in natural honey: comparison between the HPLC technique and the spectrophotometric white method, Journal of Food Science 77 (7) (2012) 784–90.
40. S. Ünüvar, Determination of 5-hydroxymethylfurfural (5-HMF) in expired pharmaceutical syrups by using HPLC-DAD method, Journal of the Turkish Chemical Society Section A: Chemistry 5 (3) (2018) 1431–1440.
41. A. Val, J. F. Huidobro, M. P. Sánchez, S. Muniategui, M. A. Fernández-Muiño, M. T. Sancho, Enzymatic determination of galactose and lactose in honey, Journal of Agricultural and Food Chemistry 46 (4) (1998) 1381–1385.
42. S. Bogdanov, T. Jurendic, R. Sieber, P. Gallmann, Honey for nutrition and health: A review, Journal of the American College of Nutrition 27 (6) (2008) 677–689.
43. A. Pascual-Maté, S. M. Osés, G. L. Marcazzan, S. Gardini, M. A. F. Muiño, M. T. Sancho, Sugar composition and sugar-related parameters of honeys from the northern Iberian Plateau, Journal of Food Composition and Analysis 74 (2018) 34–43.
44. P. Araujo, Key aspects of analytical method validation and linearity evaluation, Journal of Chromatography B 877 (23) (2009) 2224–2234.
45. E. Rozet, A. Ceccato, C. Hubert, E. Ziemons, R. Oprean, S. Rudaz, B. Boulanger, P. Hubert, Analysis of recent pharmaceutical regulatory documents on analytical method validation, Journal of Chromatography A 1158 (1-2) (2007) 111–125.
46. A. A. Ariffin, H. M. Ghazali, P. Kavousi, Validation of a HPLC method for determination of hydroxymethylfurfural in crude palm oil, Food Chemistry 154 (2014) 102–107.
47. M. I. Milani, E. L. Rossini, K. Castoldi, L. Pezza, H. R. Pezza, Paper platform for reflectometric determination of furfural and hydroxymethylfurfural in sugarcane liquor, Microchemical Journal 133 (2017) 286–292.
48. S. L. Ellison, A. Williams, Quantifying uncertainty in analytical measurement, 3rd Edition, EURACHEM/CITAC, Teddington, 2012
49. Joint FAO/WHO Codex Alimentarius Commission, Codex Alimentarius Commission Procedural Manual, 21st Edition (2013), http://www.fao.org/3/a-i3243e.pdf Accessed 28 Feb 2024
50. A. Besir, F. Yazici, M. Mortas, O. Gul, A novel spectrophotometric method based on Seliwanoff test to determine 5-(Hydroxymethyl) furfural (HMF) in honey: Development, in house validation and application, LWT 139 (2021) 110602 8 pages.
51. S. Karabournioti, P. Zervalaki, The effect of heating on honey HMF and invertase, Apiacta 36 (4) (2001) 177–181.
52. I. Turhan, N. Tetik, M. Karhan, F. Gurel, H. R. Tavukçuoğlu, Quality of honeys influenced by thermal treatment, LWT - Food Science and Technology 41 (8) (2008) 1396–1399.
53. Turkish Food Codex, Turkish Food Codex Honey Statement 2012/58 (2012), https://www.resmigazete.gov.tr/eskiler/2012/07/20120727-12.htm Accessed 28 Feb 2024