Development of a Green Analytical Method for the Assessment of Honey Quality through Reducing Sugars, Hydroxymethylfurfural and Physicochemical Parameters

Inas Yaseen Tahah

doi:10.51699/mr4bxm32

Authors

Inas Yaseen Tahah National Standards Laboratory SA

DOI:

https://doi.org/10.51699/mr4bxm32

Keywords:

Honey quality, Reducing sugars, Hydroxymethylfurfural, Green analytical chemistry, Hexacyanoferrate(III), AGREE, Chemometrics, Codex Alimentarius

Abstract

A low-cost and green spectrophotometric method was developed to be used as a routine quality control method of honey. The use of Carrez reagents, p-toluidine and Fehling's copper reagents was eliminated by simply filtering the solution through a membrane and measuring reducing sugars using a miniaturised alkaline hexacyanoferrate(III) reaction at 420 nm, and 5-hydroxymethylfurfural (HMF) at 284 nm against a sodium bisulfite reference. Moisture, pH, free acidity, electrical conductivity, ash and Pfund colour were measured with reagent-free or low-reagent procedures on the same samples. Under the optimum conditions (2.0 mmol L-1 K3[Fe(CN)6], 0.20 mol L-1 Na2CO3, 90 °C, 6 min) the sugar response was linear from 10 to 100 µg mL-1 (R2 = 0.9997), and the HMF response was linear from 1.0 to 20.0 mg L-1 (R2 = 0.9999). The limits of detection were 2.21 µg mL-1 and 0.21 mg L-1, respectively. Intra and Inter day RSD values were not more than 1.7% and 2.6% respectively and recovery ranged between 97.6% and 102.4%. Eighteen honeys from Iraqi markets and apiaries were analysed. Of the 15, all met the Codex Alimentarius and EU limits for all parameters, but three imported commercial blends failed the HMF limit (40 mg kg-1) and two of these also failed the moisture and sucrose limits. The results were in good agreement with the Lane – Eynon method and HPLC-UV method (paired t-test, p> 0.05). The procedure achieved a score of 89 on the Analytical Eco-Scale and 0.73 on AGREE in comparison to 0.49 to 0.54 for the conventional procedures. Two components accounted for 90.7% of the variance and principal component analysis discriminated heat-damaged blends from fresh local honeys.

References

[1] Codex Alimentarius Commission, Standard for Honey, CXS 12-1981, Adopted 1981, Revised 1987 and 2001, Amended 2019, Rome: FAO/WHO, 2019.

[2] J. M. Alvarez-Suarez, S. Tulipani, S. Romandini, E. Bertoli, and M. Battino, "Contribution of honey in nutrition and human health: A review," Med. J. Nutrition Metab., vol. 3, no. 1, pp. 15–23, 2010.

[3] O. Escuredo, M. Míguez, M. Fernández-González, and M. C. Seijo, "Nutritional value and antioxidant activity of honeys produced in a European Atlantic area," Food Chem., vol. 138, no. 2–3, pp. 851–856, 2013.

[4] P. M. da Silva, C. Gauche, L. V. Gonzaga, A. C. O. Costa, and R. Fett, "Honey: Chemical composition, stability and authenticity," Food Chem., vol. 196, pp. 309–323, 2016.

[5] J. W. White and L. W. Doner, "Honey composition and properties," in Beekeeping in the United States, Agriculture Handbook No. 335, Washington, DC: USDA, 1980, pp. 82–91.

[6] D. W. Ball, "The chemical composition of honey," J. Chem. Educ., vol. 84, no. 10, pp. 1643–1646, 2007.

[7] C. Pita-Calvo and M. Vázquez, "Differences between honeydew and blossom honeys: A review," Trends Food Sci. Technol., vol. 59, pp. 79–87, 2017.

[8] S. Bogdanov, K. Ruoff, and L. Persano Oddo, "Physico-chemical methods for the characterisation of unifloral honeys: A review," Apidologie, vol. 35, no. Suppl. 1, pp. S4–S17, 2004.

[9] Council of the European Union, "Council Directive 2001/110/EC of 20 December 2001 relating to honey," Off. J. Eur. Communities, vol. L10, pp. 47–52, 2002.

[10] E. Anklam, "A review of the analytical methods to determine the geographical and botanical origin of honey," Food Chem., vol. 63, no. 4, pp. 549–562, 1998.

[11] A. J. Siddiqui, S. G. Musharraf, M. I. Choudhary, and A. Rahman, "Application of analytical methods in authentication and adulteration of honey," Food Chem., vol. 217, pp. 687–698, 2017.

[12] J. H. Lane and L. Eynon, "Determination of reducing sugars by means of Fehling's solution with methylene blue as internal indicator," J. Soc. Chem. Ind., vol. 42, pp. 32T–37T, 1923.

[13] S. Bogdanov, P. Martin, and C. Lüllmann, Harmonised Methods of the International Honey Commission, Bern: Swiss Bee Research Centre, FAM, Liebefeld, 2002 (revised 2009).

[14] G. L. Miller, "Use of dinitrosalicylic acid reagent for determination of reducing sugar," Anal. Chem., vol. 31, no. 3, pp. 426–428, 1959.

[15] H. C. Hagedorn and B. N. Jensen, "Zur Mikrobestimmung des Blutzuckers mittels Ferricyanid," Biochem. Z., vol. 135, pp. 46–58, 1923.

[16] J. T. Park and M. J. Johnson, "A submicrodetermination of glucose," J. Biol. Chem., vol. 181, no. 1, pp. 149–151, 1949.

[17] S. Kowalski, M. Łukasiewicz, A. Duda-Chodak, and G. Zięć, "5-Hydroxymethyl-2-furfural (HMF): Heat-induced formation, occurrence in food and biotransformation. A review," Pol. J. Food Nutr. Sci., vol. 63, no. 4, pp. 207–225, 2013.

[18] E. Capuano and V. Fogliano, "Acrylamide and 5-hydroxymethylfurfural (HMF): A review on metabolism, toxicity, occurrence in food and mitigation strategies," LWT - Food Sci. Technol., vol. 44, no. 4, pp. 793–810, 2011.

[19] B. Fallico, M. Zappalà, E. Arena, and A. Verzera, "Effects of conditioning on HMF content in unifloral honeys," Food Chem., vol. 85, no. 2, pp. 305–313, 2004.

[20] E. Tosi, M. Ciappini, E. Ré, and H. Lucero, "Honey thermal treatment effects on hydroxymethylfurfural content," Food Chem., vol. 77, no. 1, pp. 71–74, 2002.

[21] M. I. Khalil, S. A. Sulaiman, and S. H. Gan, "High 5-hydroxymethylfurfural concentrations are found in Malaysian honey samples stored for more than one year," Food Chem. Toxicol., vol. 48, no. 8–9, pp. 2388–2392, 2010.

[22] U. M. Shapla, M. Solayman, N. Alam, M. I. Khalil, and S. H. Gan, "5-Hydroxymethylfurfural (HMF) levels in honey and other food products: Effects on bees and human health," Chem. Cent. J., vol. 12, art. no. 35, 2018.

[23] J. W. White, "Spectrophotometric method for hydroxymethylfurfural in honey," J. Assoc. Off. Anal. Chem., vol. 62, no. 3, pp. 509–514, 1979.

[24] O. Winkler, "Beitrag zum Nachweis und zur Bestimmung von Oxymethylfurfurol in Honig und Kunsthonig," Z. Lebensm. Unters. Forsch., vol. 102, no. 3, pp. 161–167, 1955.

[25] M. Zappalà, B. Fallico, E. Arena, and A. Verzera, "Methods for the determination of HMF in honey: A comparison," Food Control, vol. 16, no. 3, pp. 273–277, 2005.

[26] I. N. Pasias, I. K. Kiriakou, and C. Proestos, "HMF and diastase activity in honeys: A fully validated approach and a chemometric analysis for identification of honey freshness and adulteration," Food Chem., vol. 229, pp. 425–431, 2017.

[27] H. D. Chataway, "The determination of moisture in honey," Can. J. Res., vol. 6, no. 5, pp. 532–547, 1932.

[28] M. G. Piazza, M. Accorti, and L. Persano Oddo, "Electrical conductivity, ash, colour and specific rotatory power in Italian unifloral honeys," Apicoltura, vol. 7, pp. 51–63, 1991.

[29] V. Kaškonienė, P. R. Venskutonis, and V. Čeksterytė, "Carbohydrate composition and electrical conductivity of different origin honeys from Lithuania," LWT - Food Sci. Technol., vol. 43, no. 5, pp. 801–807, 2010.

[30] I. C. F. R. Ferreira, E. Aires, J. C. M. Barreira, and L. M. Estevinho, "Antioxidant activity of Portuguese honey samples: Different contributions of the entire honey and phenolic extract," Food Chem., vol. 114, no. 4, pp. 1438–1443, 2009.

[31] S. Gomes, L. G. Dias, L. L. Moreira, P. Rodrigues, and L. Estevinho, "Physicochemical, microbiological and antimicrobial properties of commercial honeys from Portugal," Food Chem. Toxicol., vol. 48, no. 2, pp. 544–548, 2010.

[32] M. B. Sakač, P. T. Jovanov, A. Z. Marić, L. L. Pezo, Ž. S. Kevrešan, A. R. Novaković, and N. M. Nedeljković, "Physicochemical properties and mineral content of honey samples from Vojvodina (Republic of Serbia)," Food Chem., vol. 276, pp. 15–21, 2019.

[33] P. T. Anastas and J. C. Warner, Green Chemistry: Theory and Practice, New York: Oxford University Press, 1998.

[34] S. Armenta, S. Garrigues, and M. de la Guardia, "Green analytical chemistry," TrAC Trends Anal. Chem., vol. 27, no. 6, pp. 497–511, 2008.

[35] M. Tobiszewski, A. Mechlińska, and J. Namieśnik, "Green analytical chemistry: Theory and practice," Chem. Soc. Rev., vol. 39, no. 8, pp. 2869–2878, 2010.

[36] A. Gałuszka, Z. Migaszewski, and J. Namieśnik, "The 12 principles of green analytical chemistry and the SIGNIFICANCE mnemonic of green analytical practices," TrAC Trends Anal. Chem., vol. 50, pp. 78–84, 2013.

[37] A. Gałuszka, Z. M. Migaszewski, P. Konieczka, and J. Namieśnik, "Analytical Eco-Scale for assessing the greenness of analytical procedures," TrAC Trends Anal. Chem., vol. 37, pp. 61–72, 2012.

[38] J. Płotka-Wasylka, "A new tool for the evaluation of the analytical procedure: Green Analytical Procedure Index," Talanta, vol. 181, pp. 204–209, 2018.

[39] F. Pena-Pereira, W. Wojnowski, and M. Tobiszewski, "AGREE: Analytical GREEnness metric approach and software," Anal. Chem., vol. 92, no. 14, pp. 10076–10082, 2020.

[40] W. Wojnowski, M. Tobiszewski, F. Pena-Pereira, and E. Psillakis, "AGREEprep: Analytical greenness metric for sample preparation," TrAC Trends Anal. Chem., vol. 149, art. no. 116553, 2022.

[41] International Council for Harmonisation, ICH Q2(R2): Validation of Analytical Procedures, Step 4 Guideline, Geneva: ICH, 2023.

[42] R. Bro and A. K. Smilde, "Principal component analysis," Anal. Methods, vol. 6, no. 9, pp. 2812–2831, 2014.

[43] J. H. Ward, "Hierarchical grouping to optimize an objective function," J. Am. Stat. Assoc., vol. 58, no. 301, pp. 236–244, 1963.

[44] L. Persano Oddo and R. Piro, "Main European unifloral honeys: Descriptive sheets," Apidologie, vol. 35, no. Suppl. 1, pp. S38–S81, 2004.

[45] S. Ajlouni and P. Sujirapinyokul, "Hydroxymethylfurfuraldehyde and amylase contents in Australian honey," Food Chem., vol. 119, no. 3, pp. 1000–1005, 2010.