1: total phenolic content (TPC) expressed as mg of gallic acid equivalents (GAE)/100 g; 2: total flavonoid content (TFC) expressed as mg of rutin equivalents (RUE)/100 g; *: P < 0.05; NS: non-significant.
Declarations
Acknowledgments
The authors would like to thank Mr. Dimitrios Margaritis (deceased), Department of Products & Systems Design Engineering, University of Western Macedonia, for creating the map showing the locations where the bee hives were placed for honey collection.
The authors are grateful to the Beekeeping Association of Kozani for facilitating contact with the beekeepers who provided the honey samples used in the study.
Author contributions
PM: Conceptualization, Methodology, Validation, Formal analysis, Resources, Data curation, Supervision, Project administration, Writing—original draft, Writing—review & editing. NV: Investigation. IS: Investigation. CM: Investigation. GP: Investigation. VP: Investigation. EK: Methodology, Validation, Formal analysis, Data curation, Writing—original draft, Writing—review & editing. All authors read and approved the submitted version.
Conflicts of interest
The authors declare that they have no conflicts of interest.
Ethical approval
Not applicable.
Consent to participate
Not applicable.
Consent to publication
Not applicable.
Availability of data and materials
The datasets that support the findings of this study are available from the corresponding author upon reasonable request.
Open Exploration maintains a neutral stance on jurisdictional claims in published institutional affiliations and maps. All opinions expressed in this article are the personal views of the author(s) and do not represent the stance of the editorial team or the publisher.
References
Alvarez-Suarez JM, Tulipani S, Romandini S, Bertoli E, Battino M. Contribution of honey in nutrition and human health: a review.Mediterr J Nutr Metab. 2009;3:15–23. [DOI]
Bertoncelj J, Doberšek U, Jamnik M, Golob T. Evaluation of the phenolic content, antioxidant activity and colour of Slovenian honey.Food Chem. 2007;105:822–8. [DOI]
Clearwater MJ, Revell M, Noe S, Manley-Harris M. Influence of genotype, floral stage, and water stress on floral nectar yield and composition of mānuka (Leptospermum scoparium).Ann Bot. 2018;121:501–12. [DOI] [PubMed] [PMC]
El Sohaimy S, Masry S, Shehata M. Physicochemical characteristics of honey from different origins.Ann Agric Sci. 2015;60:279–87. [DOI]
Al-Kafaween MA, Alwahsh M, Hilmi ABM, Abulebdah DH. Physicochemical Characteristics and Bioactive Compounds of Different Types of Honey and Their Biological and Therapeutic Properties: A Comprehensive Review.Antibiotics (Basel). 2023;12:337. [DOI] [PubMed] [PMC]
Viuda-Martos M, Ruiz-Navajas Y, Fernández-López J, Pérez-Alvarez JA. Functional properties of honey, propolis, and royal jelly.J Food Sci. 2008;73:R117–24. [DOI] [PubMed]
Jara-Palacios MJ, Ávila FJ, Escudero-Gilete ML, Gómez Pajuelo A, Heredia FJ, Hernanz D, et al. Physicochemical properties, colour, chemical composition, and antioxidant activity of Spanish Quercus honeydew honeys.Eur Food Res Technol. 2019;245:2017–26. [DOI]
Seraglio SKT, Silva B, Bergamo G, Brugnerotto P, Gonzaga LV, Fett R, et al. An overview of physicochemical characteristics and health-promoting properties of honeydew honey.Food Res Int. 2019;119:44–66. [DOI] [PubMed]
Pita-Calvo C, Vázquez M. Differences between honeydew and blossom honeys: A review.Trends Food Sci Technol. 2017;59:79–87. [DOI]
González-Paramás AM, García-Villanova RJ, Gómez Bárez JA, Sánchez Sánchez J, Ardanuy Albajar R. Botanical origin of monovarietal dark honeys (from heather, holm oak, pyrenean oak and sweet chestnut) based on their chromatic characters and amino acid profiles.Eur Food Res Technol. 2006;226:87–92. [DOI]
Rodopoulou M, Tananaki C, Kanelis D, Liolios V, Dimou M, Thrasyvoulou A. A chemometric approach for the differentiation of 15 monofloral honeys based on physicochemical parameters.J Sci Food Agric. 2022;102:139–46. [DOI] [PubMed]
Ntakoulas DD, Pasias IN, Tsagkaris AS, Raptopoulou KG, Raptis S, Proestos C. Chemometric differentiation of Greek chestnut and oak honeydew honeys.Int J Food Sci Technol. 2025;60:vvaf117. [DOI]
Koulis GA, Tsagkaris AS, Katsianou PA, Gialouris PP, Martakos I, Stergiou F, et al. Thorough Investigation of the Phenolic Profile of Reputable Greek Honey Varieties: Varietal Discrimination and Floral Markers Identification Using Liquid Chromatography-High-Resolution Mass Spectrometry.Molecules. 2022;27:4444. [DOI] [PubMed] [PMC]
Xagoraris M, Revelou P, Alissandrakis E, Tarantilis PA, Pappas CS. Greek Honey Authentication: Botanical Approach.Encyclopedia. 2021;1:1322–33. [DOI]
Council Directive 2001/110/EC of 20 December 2001 relating to honey [Internet].Council of the European Union; [cited 2025 Jul 20]. Available from: https://eur-lex.europa.eu/eli/dir/2001/110/oj/eng
Alvarez-Suarez JM, Gasparrini M, Forbes-Hernández TY, Mazzoni L, Giampieri F. The Composition and Biological Activity of Honey: A Focus on Manuka Honey.Foods. 2014;3:420–32. [DOI] [PubMed] [PMC]
Qi N, Zhao W, Xue C, Zhang L, Hu H, Jin Y, et al. Phenolic Acid and Flavonoid Content Analysis with Antioxidant Activity Assessment in Chinese C. pi. Shen Honey.Molecules. 2025;30:370. [DOI] [PubMed] [PMC]
Makawi SZA, Gadkariem EA, Ayoub SMH. Determination of Antioxidant Flavonoids in Sudanese Honey Samples by Solid Phase Extraction and High Performance Liquid Chromatography.J Chem. 2009;6:S429–37. [DOI]
Cianciosi D, Forbes-Hernández TY, Afrin S, Gasparrini M, Reboredo-Rodriguez P, Manna PP, et al. Phenolic Compounds in Honey and Their Associated Health Benefits: A Review.Molecules. 2018;23:2322. [DOI] [PubMed] [PMC]
Becerril-Sánchez AL, Quintero-Salazar B, Dublán-García O, Escalona-Buendía HB. Phenolic Compounds in Honey and Their Relationship with Antioxidant Activity, Botanical Origin, and Color.Antioxidants (Basel). 2021;10:1700. [DOI] [PubMed] [PMC]
Lewoyehu M, Amare M. Comparative evaluation of analytical methods for determining the antioxidant activities of honey: A review.Cogent Food Agric. 2019;5:1685059. [DOI]
Starowicz M, Ostaszyk A, Zieliński H. The Relationship between the Browning Index, Total Phenolics, Color, and Antioxidant Activity of Polish-Originated Honey Samples.Foods. 2021;10:967. [DOI] [PubMed] [PMC]
Majewska E, Drużyńska B, Derewiaka D, Ciecierska M, Pakosz P. Comparison of Antioxidant Properties and Color of Selected Polish Honeys and Manuka Honey.Foods. 2024;13:2666. [DOI] [PubMed] [PMC]
Beretta G, Granata P, Ferrero M, Orioli M, Maffei Facino R. Standardization of antioxidant properties of honey by a combination of spectrophotometric/fluorimetric assays and chemometrics.Analytica Chimica Acta. 2005;533:185–91. [DOI]
Truzzi C, Annibaldi A, Illuminati S, Finale C, Scarponi G. Determination of proline in honey: comparison between official methods, optimization and validation of the analytical methodology.Food Chem. 2014;150:477–81. [DOI] [PubMed]
Saxena S, Gautam S, Sharma A. Physical, biochemical and antioxidant properties of some Indian honeys.Food Chem. 2010;118:391–7. [DOI]
Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OG. Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan honey, as well as their radical scavenging activity.Food Chem. 2005;91:571–7. [DOI]
da Costa IF, Toro MJU. Evaluation of the antioxidant capacity of bioactive compounds and determination of proline in honeys from Pará.J Food Sci Technol. 2021;58:1900–8. [DOI] [PubMed] [PMC]
Kasiotis KM, Baira E, Iosifidou S, Manea-Karga E, Tsipi D, Gounari S, et al. Fingerprinting Chemical Markers in the Mediterranean Orange Blossom Honey: UHPLC-HRMS Metabolomics Study Integrating Melissopalynological Analysis, GC-MS and HPLC-PDA-ESI/MS.Molecules. 2023;28:3967. [DOI] [PubMed] [PMC]
Kędzierska-Matysek M, Florek M, Wolanciuk A, Skałecki P. Effect of freezing and room temperatures storage for 18 months on quality of raw rapeseed honey (Brassica napus).J Food Sci Technol. 2016;53:3349–55. [DOI] [PubMed] [PMC]
Amor SB, Mekious S, Benfekih LA, Abdellattif MH, Boussebaa W, Almalki FA, et al. Phytochemical Characterization and Bioactivity of Different Honey Samples Collected in the Pre-Saharan Region in Algeria.Life (Basel). 2022;12:927. [DOI] [PubMed] [PMC]
Ferreira IC, Aires E, Barreira JC, Estevinho LM. Antioxidant activity of Portuguese honey samples: Different contributions of the entire honey and phenolic extract.Food Chem. 2009;114:1438–43. [DOI]
Pontis JA, da Costa LAMA, da Silva SJR, Flach A. Color, phenolic and flavonoid content, and antioxidant activity of honey from Roraima, Brazil.Food Sci Technol. 2014;34:69–73. [DOI]
Pascual-Maté A, Osés SM, Fernández-Muiño MA, Sancho MT. Analysis of Polyphenols in Honey: Extraction, Separation and Quantification Procedures.Sep Purif Rev. 2017;47:142–58. [DOI]
Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent.Methods Enzymol. 1999;299:152–78. [DOI]
Bhaigyabati T, Devi PG, Bag GC. Total flavonoid content and antioxidant activity of aqueous rhizome extract of three Hedychium species of Manipur valley.Res J Pharm Biol Chem Sci. 2014;5:970–6.
Sánchez-Moreno C, Larrauri JA, Saura-Calixto F. A procedure to measure the antiradical efficiency of polyphenols.J Sci Food Agric. 1998;76:270–6. [DOI]
Pulido R, Bravo L, Saura-Calixto F. Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay.J Agric Food Chem. 2000;48:3396–402. [DOI] [PubMed]
Oliveira A, Sousa JC, Silva AC, Melo LDR, Sillankorva S. Chestnut Honey and Bacteriophage Application to Control Pseudomonas aeruginosa and Escherichia coli Biofilms: Evaluation in an ex vivo Wound Model.Front Microbiol. 2018;9:1725. [DOI] [PubMed] [PMC]
Preti R, Tarola AM. Chemometric evaluation of the antioxidant properties and phenolic compounds in Italian honeys as markers of floral origin.Eur Food Res Technol. 2022;248:991–1002. [DOI]
Wilczyńska A. Phenolic content and antioxidant activity of different types of Polish honey – A short report.Pol J Food Nutr Sci. 2010;60:309–13.
Bogdanov S, Ruoff K, Oddo LP. Physico-chemical methods for the characterisation of unifloral honeys: a review.Apidologie. 2004;35:S4–17. [DOI]
Thrasyvoulou A, Tananaki C, Goras G, Karazafiris E, Dimou M, Liolios V, et al. Legislation of honey criteria and standards.J Apic Res. 2018;57:88–96. [DOI]
Karabagias IK, Halatsi EZ, Karabournioti S, Kontakos S, Kontominas MG. Impact of physicochemical parameters, pollen grains, and phenolic compounds on the correct geographical differentiation of fir honeys produced in Greece as assessed by multivariate analyses.Int J Food Prop. 2017;20:S520–33. [DOI]
Tananaki C, Rodopoulou MA, Dimou M, Kanelis D, Liolios V. The Total Phenolic Content and Antioxidant Activity of Nine Monofloral Honey Types.Appl Sci. 2024;14:4329. [DOI]
Kolayli S, Can Z, Çakir HE, Okan OT, Yildiz O. An investigation on Trakya region Oak (Quercus spp.) honeys of Turkey: their physico-chemical, antioxidant and phenolic compounds properties.Turk J Biochem. 2018;43:362–74. [DOI]
Seijo MC, Escuredo O, Rodríguez-Flores MS. Physicochemical Properties and Pollen Profile of Oak Honeydew and Evergreen Oak Honeydew Honeys from Spain: A Comparative Study.Foods. 2019;8:126. [DOI] [PubMed] [PMC]
Vasić V, Gašić U, Stanković D, Lušić D, Vukić-Lušić D, Milojković-Opsenica D, et al. Towards better quality criteria of European honeydew honey: Phenolic profile and antioxidant capacity.Food Chem. 2019;274:629–41. [DOI] [PubMed]
Ucurum O, Tosunoglu H, Takma Ç, Birlik PM, Berber M, Kolaylı S. Distinctive properties of the pine, oak, chestnut and multifloral blossom and honeydew honeys.Eur Food Res Technol. 2024;250:1765–74. [DOI]
Wilczyńska A, Żak N. Polyphenols as the Main Compounds Influencing the Antioxidant Effect of Honey-A Review.Int J Mol Sci. 2024;25:10606. [DOI] [PubMed] [PMC]
Can Z, Yildiz O, Sahin H, Turumtay EA, Silici S, Kolayli S. An investigation of Turkish honeys: their physico-chemical properties, antioxidant capacities and phenolic profiles.Food Chem. 2015;180:133–41. [DOI] [PubMed]
Flores MSR, Escuredo O, Seijo MC. Assessment of physicochemical and antioxidant characteristics of Quercus pyrenaica honeydew honeys.Food Chem. 2015;166:101–6. [DOI] [PubMed]
Patouna A, Vardakas P, Skaperda Z, Spandidos DA, Kouretas D. Evaluation of the antioxidant potency of Greek honey from the Taygetos and Pindos mountains using a combination of cellular and molecular methods.Mol Med Rep. 2023;27:54. [DOI] [PubMed] [PMC]
de F Schaffel I, dos Santos GFS, Damm BM, de S Santos RA, Almeida JGL, Resende HC, et al. Physicochemical Properties and Antioxidant Capacity of Honey from Honey Bee (Apis mellifera): Spectrophotometric and Electrochemical Assay.ACS Omega. 2025;10:10653–61. [DOI] [PubMed] [PMC]
Lachman J, Orsák M, Hejtmánková A, Kovářová E. Evaluation of antioxidant activity and total phenolics of selected Czech honeys.LWT - Food Sci Technol. 2010;43:52–8. [DOI]
Martinello M, Mutinelli F. Antioxidant Activity in Bee Products: A Review.Antioxidants (Basel). 2021;10:71. [DOI] [PubMed] [PMC]
Karabagias IK, Dimitriou E, Kontakos S, Kontominas MG. Phenolic profile, colour intensity, and radical scavenging activity of Greek unifloral honeys.Eur Food Res Technol. 2016;242:1201–10. [DOI]
Karabagias IK, Maia M, Karabagias VK, Gatzias I, Badeka AV. Characterization of Eucalyptus, Chestnut and Heather Honeys from Portugal Using Multi-Parameter Analysis and Chemo-Calculus.Foods. 2018;7:194. [DOI] [PubMed] [PMC]
Antony SM, Han IY, Rieck JR, Dawson PL. Antioxidative effect of maillard reaction products formed from honey at different reaction times.J Agric Food Chem. 2000;48:3985–9. [DOI] [PubMed]
Bogdanov S, Lüllmann C, Martin P, von der Ohe W, Russmann H, Vorwohl G, et al. Honey quality and international regulatory standards: review by the International Honey Commission.Bee World. 1999;80:61–9. [DOI]
Cotte JF, Casabianca H, Giroud B, Albert M, Lheritier J, Grenier-Loustalot MF. Characterization of honey amino acid profiles using high-pressure liquid chromatography to control authenticity.Anal Bioanal Chem. 2004;378:1342–50. [DOI] [PubMed]
Kečkeš J, Trifković J, Andrić F, Jovetić M, Tešić Z, Milojković-Opsenica D. Amino acids profile of Serbian unifloral honeys.J Sci Food Agric. 2013;93:3368–76. [DOI] [PubMed]
Iglesias MT, de Lorenzo C, del Carmen Polo M, Martín-Alvarez PJ, Pueyo E. Usefulness of amino acid composition to discriminate between honeydew and floral honeys. Application to honeys from a small geographic area.J Agric Food Chem. 2004;52:84–9. [DOI] [PubMed]
Descamps C, Quinet M, Jacquemart A. Climate Change-Induced Stress Reduce Quantity and Alter Composition of Nectar and Pollen From a Bee-Pollinated Species (Borago officinalis, Boraginaceae).Front Plant Sci. 2021;12:755843. [DOI] [PubMed] [PMC]
Chua LS, Lee JY, Chan GF. Honey protein extraction and determination by mass spectrometry.Anal Bioanal Chem. 2013;405:3063–74. [DOI] [PubMed]
Bogdanov S. Honey Composition.In: The Honey Book. Bee Product Science; 2017.
Sommano SR, Bhat FM, Wongkeaw M, Sriwichai T, Sunanta P, Chuttong B, et al. Amino Acid Profiling and Chemometric Relations of Black Dwarf Honey and Bee Pollen.Front Nutr. 2020;7:558579. [DOI] [PubMed] [PMC]
da Silva PM, Gauche C, Gonzaga LV, Costa ACO, Fett R. Honey: Chemical composition, stability and authenticity.Food Chem. 2016;196:309–23. [DOI] [PubMed]
Kędzierska-Matysek M, Stryjecka M, Teter A, Skałecki P, Domaradzki P, Florek M. Relationships between the Content of Phenolic Compounds and the Antioxidant Activity of Polish Honey Varieties as a Tool for Botanical Discrimination.Molecules. 2021;26:1810. [DOI] [PubMed] [PMC]
Perna A, Simonetti A, Intaglietta I, Sofo A, Gambacorta E. Metal content of southern Italy honey of different botanical origins and its correlation with polyphenol content and antioxidant activity.Int J of Food Sci Tech. 2012;47:1909–17. [DOI]
Gheldof N, Engeseth NJ. Antioxidant capacity of honeys from various floral sources based on the determination of oxygen radical absorbance capacity and inhibition of in vitro lipoprotein oxidation in human serum samples.J Agric Food Chem. 2002;50:3050–5. [DOI] [PubMed]
Shamsudin S, Selamat J, Sanny M, Abd Razak S, Jambari NN, Mian Z, et al. Influence of origins and bee species on physicochemical, antioxidant properties and botanical discrimination of stingless bee honey.Int J Food Prop. 2019;22:239–64. [DOI]
