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Changes in the antioxidative properties of honeys during their fermentation

  • Bronisław K. Głód EMAIL logo and Paweł Piszcz
Published/Copyright: May 19, 2021
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Open Chemistry
From the journal Open Chemistry Volume 19 Issue 1

Abstract

The aim of this preliminary study was to investigate whether the fermentation process affects the antioxidative properties of honeys. Therefore, the total antioxidant potential (TAP) of eleven meads was examined. TAPs were estimated using DPPH, hydroxyl radicals, and amperometric measurements in the flow-through HPLC system. The results were correlated with the total content of polyphenols and anthocyanins. Additionally, the concentrations of ethanol, sugars, and hydrogen peroxide were measured. The influence of side reactions and the presence of hydrogen peroxide, glucose, ethanol, and anthocyanins on the free radicals generation were also tested. The use of HPLC electrochemical detection for TAP measurements is particularly preferred. Depending on the potential used, different antioxidants can be measured. It turned out that fruit and species meads are characterized by much higher TAPs measured at high potential of the working electrode than at lower potential due to the high concentration of weak antioxidants which do not provide signals at low potentials. It was found that dark honeys (buckwheat, honeydew) are characterized by much higher TAP values than light ones (acacia, linden, multi-floral). The concentration of anthocyanins decreased during fermentation. They are removed together with the sediment.

Keywords: mead; honey; total antioxidant potential; high-performance liquid chromatography; free radicals; Fenton reaction

1 Introduction

Metabolic processes in every living organism are carried out to produce a sufficient amount of energy for the body, to grow and function properly. Most of the metabolic processes are sustained by oxygen [1]. Unfortunately, this leads to the formation of reactive oxygen species (ROS), among them free radicals (FR) [2]. Radicals nonspecifically react with all cell components and compounds by oxidizing them and cause cell apoptosis [3]. The human body itself can eliminate the free radicals by producing endogenous free radicals scavengers and/or antioxidants. Antioxidants can also be administered by food, drink, cosmetics, or injections [4].

In the literature, various total antioxidant potential (TAP) assays are described [5,7,14]. The classical photometric or fluorymetric assays use various stable radicals or their generators. The sample oxidation is measured using different analytical assays (photometric, fluorymetric, luminescence measurements, MS, HPLC, GC, thermogravimetric, etc.). The TAP measure is in this case the inhibition time, induced by the sample. It seems that natural methods of examination of oxidation process should involve electroanalytical techniques. As a matter of fact, we have elaborated such voltammetric assay [16]. The use of volametry in HPLC detection significantly decreases the detection limit due to the high convection currents and the lack of capacitive currents.

The aim and the novelty of the paper are to examine the influence of the fermentation on changes in the antioxidative properties of honeys/meads.

2 Materials and methods

2.1 Reagents

Methanol (pure for chromatography), buffered saline tablets pH 7.4, DPPH (2,2-diphenyl-1-picrylhydrazyl), iron(ii) sulfate(vi), and 4-hydroxybenzoic acid were obtained from Sigma-Aldrich (St. Louis, Missouri, USA). Other reagents were at least of analytical-reagent grade and were used without further purification.

2.2 Instrumentation

HPLC measurements were performed using a chromatograph (Knauer, Berlin, Germany) equipped with a Cosmosil 5 microns, 4.6 × 150 mm, 5C18-MS-II column. Photometric measurements were performed using Helios Epsilon spectrophotometer (Thermo Fisher Scientific, Rochester, USA).

2.3 Materials

The antioxidative properties of eleven samples of meads (five fruit, one herbal, and five species) were investigated. Honeys were purchased from apiaries situated in two localities of Siedlce vicinity, Mazowieckie Voivodeship, Poland. They were prepared as mead trójniak, made using two units of water for one unit of honey. Fruit and herbal meads were prepared on the basis of linden honey.

The honey wort was boiled for 60 min, and then foam was removed from the surface. The wort was then transferred to the fermentation balloon. Mother yeast, 4 g of citric acid, and 0.5 g of diammonium phosphate were added. The balloon was closed with a distillation tube, into which a tablespoon of water was poured and set aside at room temperature (about 22°C). After six days, a baker’s yeast was added. On 41 day, sediments were collected. Samples were collected at various stages of production on day 0, 6, 13, 20, 27, 34, 41, 57, 69, 85, and 120.

2.4 Methods

Determination of the TAP related to the hydroxyl radical, TAPOH, is based on the generation of hydroxyl radicals in the Fenton reaction as it was described in [5]. The oxidation of p-HBA by Fe(iii) ions in the presence of glucose and ethanol was also investigated [6]. TAP assay based on HPLC-ED measurements, TAPED, and related to DPPH radicals, TAPDPPH, was described by us earlier [7]. The total polyphenol content, PF, was determined by the colorimetric method with Folin-Ciocialteu (FC) reagent [8]. Determination of anthocyanins was carried out using two analytical assays, photometric and HPLC [9]. Ethyl alcohol was determined by pycnometric measurement of distillate density according to PN-A-79120/04 [10]. Determination of sugar content has been performed by the refractometric method according to PN-A-74252 [11].

  1. Ethical approval: Not applicable, the conducted research is not related to either human or animal use.

3 Results and discussion

The changes of averaged TAPOH, TAPDPPH, and PF relative (100% refers to the highest value measured with a given assay) values of overall tested honeys during their fermentation are presented in Figure 1. For most of honeys, the TAPs decreased during the first 6 days. After this time, the fermentation accelerated. Similar changes were observed for TAPDPPH and PF. Because OH radicals react nearly with all chemical compounds (among them with the alcohols and sugars), the TAPOH for all meads are very similar (Figure 2).

Figure 1 Changes of relative TAPOH (●), PF (▲), and TAPDPPH (■) [%] values during fermentation of honeys/meads.
Figure 1

Changes of relative TAPOH (●), PF (▲), and TAPDPPH (■) [%] values during fermentation of honeys/meads.

Figure 2 Correlation between relative (from 0 to 100%) concentration of ethanol (♦), sugar (-), PF (▲), TAPDPPH (■), TAPOH (●), TAPED-1V (×), and TAPED-0.4V (+) of the produced meads.
Figure 2

Correlation between relative (from 0 to 100%) concentration of ethanol (♦), sugar (-), PF (▲), TAPDPPH (■), TAPOH (●), TAPED-1V (×), and TAPED-0.4V (+) of the produced meads.

The measurements of TAPED were performed at the potential range 0.2 to 1 V. The oxidation potential defines the antioxidant strength, while the electric current – the antioxidant concentration. The fruit meads (elderberry, raspberry, bilberry) are characterized by high antioxidant potential measured at 1 V (Figure 2). However, only one of them (melissa) contains strong antioxidant (big peaks at 0.4 V). Species honey is characterized by much bigger total surface area at 1 V potential than at lower potentials as the high concentration of weak antioxidants does not provide signals at low potentials. It turned out that dark honeys (buckwheat, honeydew) are characterized by much higher TAP values than light honeys (acacia, linden, multi-floral) [12].

The concentration of anthocyanins in fruit honeys ranged from 0.17 mg/100 g for bilberry up to 2.5 mg/100 g for blueberry honey. It turned out that their concentration decreased 2 times during fermentation. The sediments collected from honey had an intense red-blue color, characteristic of anthocyanins. It means that most of the anthocyanins are removed together with the sediment, and only a small percentage of them remain in the meads.

During the honey fermentation, sugar is converted into alcohol. The highest decrease of sugar concentration during fermentation (conversion from honey to mead) has been observed for a blueberry, the smallest for the chokeberry honey/mead. The level of alcohol concentration in the obtained meads varied from 8 to 13%.

Honeys contain hydrogen peroxide that explains their antibacterial properties [13]. The original method of its determination was elaborated. It was based on the Fenton reaction (honey was used instead of H2O2) and was characterized by the detection limit of 1.2 × 10−5%, for S/N = 3. It turned out that the hydrogen peroxide in investigated honeys and meads was below the detection limit.

The TAP is equal to the sum of products from the concentrations of all antioxidants in the sample and their antioxidant power [14]. It turned out that TAPOH values are not additive. Hydroxyl radical reacts with sugars and alcohol. On the other side, it was found that glucose and ethanol may participate in the formation of hydroxyl radicals [15].

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Acknowledgments

This work was performed within the framework of the research project no. 20/20/B of Siedlce University of Natural Sciences and Humanities. We would like to thank P. Wantusiak and A. Szabrański for their assistance in performing the experiments.

  1. Funding information: This work was performed within the framework of the research project no. 20/20/B of Siedlce University of Natural Sciences and Humanities.

  2. Author contributions: B. K. G. – conceptualization, formal analysis, methodology, writing original draft, visualization; P. P. – experimental analysis, processing of results, writing original draft, visualization, editing.

  3. Conflict of interest: Authors declare no conflict of interest.

  4. Data availability statement: All data generated or analyzed during this study are included in this published article.

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Received: 2020-11-22
Revised: 2021-02-20
Accepted: 2021-03-09
Published Online: 2021-05-19

© 2021 Bronisław K. Głód and Paweł Piszcz, published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/chem-2021-0041
Keywords for this article
mead; honey; total antioxidant potential; high-performance liquid chromatography; free radicals; Fenton reaction
Creative Commons
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  25. Sodium borohydride (NaBH4) as a high-capacity material for next-generation sodium-ion capacitors
  26. Aroma components of tobacco powder from different producing areas based on gas chromatography ion mobility spectrometry
  27. The effects of salinity on changes in characteristics of soils collected in a saline region of the Mekong Delta, Vietnam
  28. Synthesis, properties, and activity of MoVTeNbO catalysts modified by zirconia-pillared clays in oxidative dehydrogenation of ethane
  29. Synthesis and crystal structure of N,N′-bis(4-chlorophenyl)thiourea N,N-dimethylformamide
  30. Quantitative analysis of volatile compounds of four Chinese traditional liquors by SPME-GC-MS and determination of total phenolic contents and antioxidant activities
  31. A novel separation method of the valuable components for activated clay production wastewater
  32. On ve-degree- and ev-degree-based topological properties of crystallographic structure of cuprite Cu2O
  33. Antihyperglycemic effect and phytochemical investigation of Rubia cordifolia (Indian Madder) leaves extract
  34. Microsphere molecularly imprinted solid-phase extraction for diazepam analysis using itaconic acid as a monomer in propanol
  35. A nitric oxide-releasing prodrug promotes apoptosis in human renal carcinoma cells: Involvement of reactive oxygen species
  36. Machine vision-based driving and feedback scheme for digital microfluidics system
  37. Study on the application of a steam-foam drive profile modification technology for heavy oil reservoir development
  38. Ni–Ru-containing mixed oxide-based composites as precursors for ethanol steam reforming catalysts: Effect of the synthesis methods on the structural and catalytic properties
  39. Preparation of composite soybean straw-based materials by LDHs modifying as a solid sorbent for removal of Pb(ii) from water samples
  40. Synthesis and spectral characterizations of vanadyl(ii) and chromium(iii) mixed ligand complexes containing metformin drug and glycine amino acid
  41. In vitro evaluation of lactic acid bacteria with probiotic activity isolated from local pickled leaf mustard from Wuwei in Anhui as substitutes for chemical synthetic additives
  42. Utilization and simulation of innovative new binuclear Co(ii), Ni(ii), Cu(ii), and Zn(ii) diimine Schiff base complexes in sterilization and coronavirus resistance (Covid-19)
  43. Phosphorylation of Pit-1 by cyclin-dependent kinase 5 at serine 126 is associated with cell proliferation and poor prognosis in prolactinomas
  44. Molecularly imprinted membrane for transport of urea, creatinine, and vitamin B12 as a hemodialysis candidate membrane
  45. Optimization of Murrayafoline A ethanol extraction process from the roots of Glycosmis stenocarpa, and evaluation of its Tumorigenesis inhibition activity on Hep-G2 cells
  46. Highly sensitive determination of α-lipoic acid in pharmaceuticals on a boron-doped diamond electrode
  47. Synthesis, chemo-informatics, and anticancer evaluation of fluorophenyl-isoxazole derivatives
  48. In vitro and in vivo investigation of polypharmacology of propolis extract as anticancer, antibacterial, anti-inflammatory, and chemical properties
  49. Topological indices of bipolar fuzzy incidence graph
  50. Preparation of Fe3O4@SiO2–ZnO catalyst and its catalytic synthesis of rosin glycol ester
  51. Construction of a new luminescent Cd(ii) compound for the detection of Fe3+ and treatment of Hepatitis B
  52. Investigation of bovine serum albumin aggregation upon exposure to silver(i) and copper(ii) metal ions using Zetasizer
  53. Discoloration of methylene blue at neutral pH by heterogeneous photo-Fenton-like reactions using crystalline and amorphous iron oxides
  54. Optimized extraction of polyphenols from leaves of Rosemary (Rosmarinus officinalis L.) grown in Lam Dong province, Vietnam, and evaluation of their antioxidant capacity
  55. Synthesis of novel thiourea-/urea-benzimidazole derivatives as anticancer agents
  56. Potency and selectivity indices of Myristica fragrans Houtt. mace chloroform extract against non-clinical and clinical human pathogens
  57. Simple modifications of nicotinic, isonicotinic, and 2,6-dichloroisonicotinic acids toward new weapons against plant diseases
  58. Synthesis, optical and structural characterisation of ZnS nanoparticles derived from Zn(ii) dithiocarbamate complexes
  59. Presence of short and cyclic peptides in Acacia and Ziziphus honeys may potentiate their medicinal values
  60. The role of vitamin D deficiency and elevated inflammatory biomarkers as risk factors for the progression of diabetic nephropathy in patients with type 2 diabetes mellitus
  61. Quantitative structure–activity relationship study on prolonged anticonvulsant activity of terpene derivatives in pentylenetetrazole test
  62. GADD45B induced the enhancing of cell viability and proliferation in radiotherapy and increased the radioresistance of HONE1 cells
  63. Cannabis sativa L. chemical compositions as potential plasmodium falciparum dihydrofolate reductase-thymidinesynthase enzyme inhibitors: An in silico study for drug development
  64. Dynamics of λ-cyhalothrin disappearance and expression of selected P450 genes in bees depending on the ambient temperature
  65. Identification of synthetic cannabinoid methyl 2-{[1-(cyclohexylmethyl)-1H-indol-3-yl] formamido}-3-methylbutanoate using modern mass spectrometry and nuclear magnetic resonance techniques
  66. Study on the speciation of arsenic in the genuine medicinal material honeysuckle
  67. Two Cu(ii)-based coordination polymers: Crystal structures and treatment activity on periodontitis
  68. Conversion of furfuryl alcohol to ethyl levulinate in the presence of mesoporous aluminosilicate catalyst
  69. Review Articles
  70. Hsien Wu and his major contributions to the chemical era of immunology
  71. Overview of the major classes of new psychoactive substances, psychoactive effects, analytical determination and conformational analysis of selected illegal drugs
  72. An overview of persistent organic pollutants along the coastal environment of Kuwait
  73. Mechanism underlying sevoflurane-induced protection in cerebral ischemia–reperfusion injury
  74. COVID-19 and SARS-CoV-2: Everything we know so far – A comprehensive review
  75. Challenge of diabetes mellitus and researchers’ contributions to its control
  76. Advances in the design and application of transition metal oxide-based supercapacitors
  77. Color and composition of beauty products formulated with lemongrass essential oil: Cosmetics formulation with lemongrass essential oil
  78. The structural chemistry of zinc(ii) and nickel(ii) dithiocarbamate complexes
  79. Bioprospecting for antituberculosis natural products – A review
  80. Recent progress in direct urea fuel cell
  81. Rapid Communications
  82. A comparative morphological study of titanium dioxide surface layer dental implants
  83. Changes in the antioxidative properties of honeys during their fermentation
  84. Erratum
  85. Erratum to “Corrosion study of copper in aqueous sulfuric acid solution in the presence of (2E,5E)-2,5-dibenzylidenecyclopentanone and (2E,5E)-bis[(4-dimethylamino)benzylidene]cyclopentanone: Experimental and theoretical study”
  86. Erratum to “Modified TDAE petroleum plasticiser”
  87. Corrigendum
  88. Corrigendum to “A nitric oxide-releasing prodrug promotes apoptosis in human renal carcinoma cells: Involvement of reactive oxygen species”
  89. Special Issue on 3rd IC3PE 2020
  90. Visible light-responsive photocatalyst of SnO2/rGO prepared using Pometia pinnata leaf extract
  91. Antihyperglycemic activity of Centella asiatica (L.) Urb. leaf ethanol extract SNEDDS in zebrafish (Danio rerio)
  92. Selection of oil extraction process from Chlorella species of microalgae by using multi-criteria decision analysis technique for biodiesel production
  93. Special Issue on the 14th Joint Conference of Chemistry (14JCC)
  94. Synthesis and in vitro cytotoxicity evaluation of isatin-pyrrole derivatives against HepG2 cell line
  95. CO2 gas separation using mixed matrix membranes based on polyethersulfone/MIL-100(Al)
  96. Effect of synthesis and activation methods on the character of CoMo/ultrastable Y-zeolite catalysts
  97. Special Issue on Electrochemical Amplified Sensors
  98. Enhancement of graphene oxide through β-cyclodextrin composite to sensitive analysis of an antidepressant: Sulpiride
  99. Investigation of the spectroelectrochemical behavior of quercetin isolated from Zanthoxylum bungeanum
  100. An electrochemical sensor for high sensitive determination of lysozyme based on the aptamer competition approach
  101. An improved non-enzymatic electrochemical sensor amplified with CuO nanostructures for sensitive determination of uric acid
  102. Special Issue on Applied Biochemistry and Biotechnology 2020
  103. Fast discrimination of avocado oil for different extracted methods using headspace-gas chromatography-ion mobility spectroscopy with PCA based on volatile organic compounds
  104. Effect of alkali bases on the synthesis of ZnO quantum dots
  105. Quality evaluation of Cabernet Sauvignon wines in different vintages by 1H nuclear magnetic resonance-based metabolomics
  106. Special Issue on the Joint Science Congress of Materials and Polymers (ISCMP 2019)
  107. Diatomaceous Earth: Characterization, thermal modification, and application
  108. Electrochemical determination of atenolol and propranolol using a carbon paste sensor modified with natural ilmenite
  109. Special Issue on the Conference of Energy, Fuels, Environment 2020
  110. Assessment of the mercury contamination of landfilled and recovered foundry waste – a case study
  111. Primary energy consumption in selected EU Countries compared to global trends
  112. Modified TDAE petroleum plasticiser
  113. Use of glycerol waste in lactic acid bacteria metabolism for the production of lactic acid: State of the art in Poland
  114. Topical Issue on Applications of Mathematics in Chemistry
  115. Theoretical study of energy, inertia and nullity of phenylene and anthracene
  116. Banhatti, revan and hyper-indices of silicon carbide Si2C3-III[n,m]
  117. Topical Issue on Agriculture
  118. Occurrence of mycotoxins in selected agricultural and commercial products available in eastern Poland
  119. Special Issue on Ethnobotanical, Phytochemical and Biological Investigation of Medicinal Plants
  120. Acute and repeated dose 60-day oral toxicity assessment of chemically characterized Berberis hispanica Boiss. and Reut in Wistar rats
  121. Phytochemical profile, in vitro antioxidant, and anti-protein denaturation activities of Curcuma longa L. rhizome and leaves
  122. Antiplasmodial potential of Eucalyptus obliqua leaf methanolic extract against Plasmodium vivax: An in vitro study
  123. Prunus padus L. bark as a functional promoting component in functional herbal infusions – cyclooxygenase-2 inhibitory, antioxidant, and antimicrobial effects
  124. Molecular and docking studies of tetramethoxy hydroxyflavone compound from Artemisia absinthium against carcinogens found in cigarette smoke
  125. Special Issue on the Joint Science Congress of Materials and Polymers (ISCMP 2020)
  126. Preparation of cypress (Cupressus sempervirens L.) essential oil loaded poly(lactic acid) nanofibers
  127. Influence of mica mineral on flame retardancy and mechanical properties of intumescent flame retardant polypropylene composites
  128. Production and characterization of thermoplastic elastomer foams based on the styrene–ethylene–butylene–styrene (SEBS) rubber and thermoplastic material
  129. Special Issue on Applied Chemistry in Agriculture and Food Science
  130. Impact of essential oils on the development of pathogens of the Fusarium genus and germination parameters of selected crops
  131. Yield, volume, quality, and reduction of biotic stress influenced by titanium application in oilseed rape, winter wheat, and maize cultivations
  132. Influence of potato variety on polyphenol profile composition and glycoalcaloid contents of potato juice
  133. Carryover effect of direct-fed microbial supplementation and early weaning on the growth performance and carcass characteristics of growing Najdi lambs
  134. Special Issue on Applied Biochemistry and Biotechnology (ABB 2021)
  135. The electrochemical redox mechanism and antioxidant activity of polyphenolic compounds based on inlaid multi-walled carbon nanotubes-modified graphite electrode
  136. Study of an adsorption method for trace mercury based on Bacillus subtilis
  137. Special Issue on The 1st Malaysia International Conference on Nanotechnology & Catalysis (MICNC2021)
  138. Mitigating membrane biofouling in biofuel cell system – A review
  139. Mechanical properties of polymeric biomaterials: Modified ePTFE using gamma irradiation
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