Determination of physicochemical properties of multifloral honeys stored in different containers

In comparison with the initial values, acidity and 5‐(hydroxy‐methyl)‐2‐furaldehyde (HMF) values of all honey samples increased during storage. The total phenol contents of honey were decreased from 286.2 to 108.6 mg GAE/kg during 9 months of storage in white colored container. In addition, total phenol contents of honey sample stored in tin container ranged from 294.7 to 258.6 mg GAE/kg (p < .05). While radical scavenging activity values of honey samples stored in amber colored container during storage are measured as 78.6 g/L, IC₅₀, 77.9 g/L, IC50, and 76.1 g/L, IC₅₀, these activity values were determined as 84.9 g/L, IC₅₀, 80.7 g/L, IC_50, and 75.2 g/L, IC₅₀ in tin container (p < .05), respectively. While acidity values of honey samples stored in white bottle range from 23.6 (3th month) to 25.7% (9th month), it varied from 20.6 (3th month) to 21.6% (9th month) in honey sample stored in amber colored container during storage.

Practical applications

Different honeys are produced in different parts of Turkey. The known honeys are pine, citrus, and chestnut honey. Various flower honeys are produced in these and other regions. HMF formation is considered as a bed result of storage and heating.     

Kinetics of 3-deoxy-D-erythro-hexos-2-ulose in unifloral honeys

In this study, for the first time, the amount of 3-deoxy-D-erythro-hexos-2-ulose (3-DG) in fresh citrus and chestnut honeys was determined. 3-DG was measured as the corresponding quinoxalines after derivatization with orthophenylenediamine using reverse-phase high-performance liquid chromatography (RP-HPLC). Notwithstanding the freshness of the samples, high levels of 3-DG were detected in both honeys. The comparison of 3-DG and 5-hydroxymethylfurfural (HMF) concentrations, which was also quantified by RP-HPLC, showed that citrus honeys had the lowest amount of 3-DG (107 mg/kg) and the highest of HMF (16.7 mg/kg), while chestnut honeys had the opposite (398 and 1.2 mg/kg, respectively). During thermal treatment, different 3-DG and HMF trends were highlighted between the citrus and chestnut honeys; at the end, 3-DG formation was more favored with respect to HMF formation. Moreover, in citrus honeys, a good correlation between 3-DG and HMF levels was observed, which was not found in chestnut honeys, suggesting a role of the high pH values of these honeys on the degradation routes. The kinetic analysis showed the highest k value for 3-DG and HMF formation in chestnut and citrus honeys, respectively. The lowest Ea values related to 3-DG formation and the highest to HMF formation, indicating that the key intermediate 3-DG is easily formed at low temperatures, whilst the formation of HMF requires higher temperatures. For this reason, 3-DG seems to be an aging index rather than a thermal index and its use, at least for honeys at high pH values, together with HMF, could improve their quality assessment.     

PREDICTION OF HONEY SHELF LIFE

Fourteen commercial honey samples of different botanical origin (acacia, chestnut, citrus, eucalyptus, multifloral) were stored for up to 18 months at room temperature. Both 5-hydroxymethylfurfural (HMF) and diastase were evaluated and kinetics carried out. The highest HMF increase was in citrus and eucalyptus honeys at 3 mg/kg/month; the lowest in chestnut at 0.256 mg/kg/month. The highest diastase deactivation was in eucalyptus honey at 0.485 DU/kg/month; the lowest was in chestnut at 0.258 DU/kg/month. Honey shelf life was estimated for both indices, HMF and diastase, using a Bayesian approach. The results show that commercial honey shelf life depends on botanical origin as well as processing. Except chestnut, all other honeys showed shorter shelf lives than the declared one (usually 36 months). The shortest values, 15 months, were for citrus and eucalyptus honeys. The longest, 20 months, was for acacia and multifloral honeys.

PRACTICAL APPLICATIONS

Both kinetic equations and results of simulations can be used to estimate, for quality control purposes as well as regulation requirements, the most probable value of shelf life for a specific honey.     

Kinetics of 5-hydroxymethylfurfural formation in chinese acacia honey during heat treatment

In this paper, the 5-hydroxymethylfurfural (HMF) contents of the acacia honeys after heat treatment were determined by HPLC; the kinetics of HMF formation was also investigated. The HMF content of acacia honey was 0.38±0.01mg/kg, but rapidly increased to the maximum of 18,320.07±14.29 mg/kg at 190°C and decreased to 1,180.24±6.54 mg/kg at 230°C after heating. The HMF content increased gradually in honey samples heated in 12 h at the temperature of 80, 100, and 120°C. The apparent frequency constant was k _f =3.91×10²⁵/h, and the apparent activation energy was E _a=173.10 kJ/mol. The HMF content in the acacia honey was related to honey composition, heating temperature, and time, and the HMF formation could also be related to the initial honey pH.     

Investigating 5-hydroxymethylfurfural formation kinetic and antioxidant activity in heat treated honey from different floral sources

The quality and biochemical properties of honey are affected by heating or during storage period. The most important biochemical reaction that occurs in this process is the Maillard reaction. HMF (5-hydroxymethylfurfural) is one of the major intermediate products in the Maillard reaction that can lead to quality reduction in heated honey. In this study, the effect of heating on the antioxidant activity, and colour values as Maillard reaction indicators of three different botanical honeys were investigated; the HMF formation was also determined. Temperatures of 50, 70 and 80 °C were applied on the honeys between 0 and 48 h. Total phenolic content, ferric reducing/antioxidant power and scavenging of 2,2-diphenyl-1-picrylhydrazyl free radical assays are used to determine the antioxidant capacity. Results showed that the formation of HMF and the antioxidant properties of honeys were significantly increased during the heating process. Pure HMF compound also showed lower antioxidant activity. The formation of HMF has higher degree of linearity in the fit of the zero order reaction and also it was the highest in the chestnut honey. Furthermore, it was found that as the biochemical value of the honeys increased, the HMF formation decelerated.     

HPLC flavonoid profiles as markers for the botanical origin of European unifloral honeys

The HPLC phenolic profiles of 52 selected unifloral honey samples produced in Europe were analysed to detect possible markers for the floral origin of the different honeys. Lime‐tree (five markers), chestnut (five markers), rapeseed (one marker), eucalyptus (six markers) and heather (three markers) honeys had specific markers with characteristic UV spectra. In addition, the flavanone hesperetin was confirmed as a marker for citrus honey, as well as kaempferol for rosemary honey and quercetin for sunflower honey. Abscisic acid, which had been reported to be a possible marker for heather honey, was also detected in rapeseed, lime‐tree and acacia honeys. Ellagic acid in heather honey and the hydroxycinnamates caffeic, p‐coumaric and ferulic acids in chestnut, sunflower, lavender and acacia honeys were also detected. The characteristic propolis‐derived flavonoids pinocembrin, pinobanksin and chrysin were present in most samples in variable amounts. © 2001 Society of Chemical Industry     

Hesperetin: A marker of the floral origin of citrus honey

Seventeen flavonoid aglycones were identified in various experimental and commercial citrus honey samples by HPLC analysis. The flavanone hesperetin was detected in all samples. This flavanone was not detected in any of the honey samples, from diverse floral origin (including rosemary, lavender, sunflower, almond, sweet chestnut, white clover, Erisarum, Robinia, Rhododendron, Tilia, Prosopis, Eucalyptus and Calluna honeys) previously investigated. The analysis of the flavonoids present in orange nectar revealed that the flavanone hesperidin (hesperetin-7-rutinoside) was the major flavonoid detected and, therefore, this should be the main source of the hesperetin found in citrus honey. Hesperetin should be produced by hydrolysis of hesperidin by the bee enzymes present in honey. These results suggest that hesperetin could be used as a marker for the botanical origin of citrus honey.     

Effect of postharvest storage conditions on the colour and freshness parameters of raw honey

The influence of temperature (25, 30, 35 and 40 °C) and storage time (up to 62 or 104 days) on colour; hydroxymethyl furfural (HMF) content; and diastase index (ID) of citrus, rosemary, eucalyptus, polyfloral and honeydew honey was evaluated. Decreases in luminosity, whiteness index, hue and chroma with the increase in temperature and storage time were observed to different extents depending on the botanical origin of the honey. At all temperatures, the highest values of chromatic parameters throughout the whole period of storage corresponded to citrus honey. Honeydew honey showed the lowest values and the fewest colour differences. Principal component analysis explained 89% of the variations. PC1 (73%) separated honey types at lower temperatures in terms of colour parameters. PC2 (16%) described variations in HMF and ID in relation to storage conditions. Honey type had a much greater influence on HMF, ID and colour than storage conditions. The evaluated postharvest treatments did not alter the characteristic colour of each type of honey as long as the commercial requirements for freshness parameters were met.     

Quality of honeys influenced by thermal treatment

Honey producers have been heating honeys at mild temperatures below 100 °C chiefly in order to prevent post-bottling crystallization. In this study, we aimed to determine the effects of thermal treatment on the HMF content of honeydew and floral honey during the isothermal heating process at mild temperatures. Water content, formol number, total acidity, pH value and minerals were also determined in both honey types as their characteristics differ with composition, which is able to affect the rate of HMF formation. Potassium content and pH value were found as the distinguishing properties and both were greater in honeydew honey than in floral honey (p<0.01). Honeydew and floral honey samples were heated at 75, 90 and 100 °C for 15-90 min and analysed for HMF content by HPLC-RP. The Arrhenius model was used to calculate reaction rate constants and activation energies which were found to be different for each of the honey types. Heating at 90 °C for up to 90 min in floral honeys and up to 75 min in honeydew honeys did not cause a significant increase of HMF and not exceed the threshold level of 40 mg kg⁻¹. Our results show that the excessive HMF content might be related to primitive storage conditions rather than overheating.     

Kinetic Parameter Estimation for Quality Change during Continuous Thermal Processing of Grapefruit Juice

Reaction order, activation energy (E_a) and average rate constant (k_av) were evaluated by dynamic modeling for color degradation of grapefruit juice during thermal processing. Simulated aseptic conditions were: 85–135°C, 40–175 set processing time, and average velocities 5–30 cm/set. Quality was assessed by nonenzymatic browning index (NEBI) and S-hydroxymethyl-furfural (HMF). Average values for reaction-order and energy of activation were: 0 and 21.4 kcal/g-mol (NEBI) and 0.3 and 23.2 kcal/g-mol (HMF), respectively. These values were compared with those contained in the deformed ellipsoid which define the 90% joint confidence interval of all possible parameter combinations. Based on initial and final concentrations, the method can predict effects of processing on liquid foods.     

Effect of storage on nonenzymatic browning reactions in carob pekmez

Carob pekmez was stored at 5, 25, 35 and 45 °C for studying the reaction kinetics of nonenzymatic browning reactions. Hydroxymethylfurfural (HMF) formation, browning index (A₄₂₀) and CIE (International Commission on Illumination) colour parameters were analysed to evaluate nonenzymatic browning reactions. HMF formation and A₄₂₀ values increased linearly with the storage time and temperature and both followed zero‐order reaction. No fitting model was found for the changes in visual CIE parameters. The dependence of rate constant of nonenzymatic reactions on temperature was represented by an Arrhenius equation and the activation values were found as 114.87 kJ mol^?1 and 86.62 kJ mol^?1 for HMF formation and A₄₂₀ values, respectively. The excellent linear correlations (r = 0.728–0.99) among colour parameters, browning index and HMF were found.     

Combined use of HMF and furosine to assess fresh honey quality

BACKGROUND: The quality of honey can be affected by practices such as adulteration, inadequate storage or the application of severe heat treatments. Because hydroxymehylfurfural (HMF) is an indicator of honey freshness and furosine (ε‐2‐furoylmethyl lysine) has proved to be a useful chemical indicator of the progress of the Maillard reaction in foods, the aim of this work was to assess their usefulness as indicators of fresh honey quality. The effect of heat treatment, storage and adulteration with different types of syrups on HMF and furosine content has been studied. RESULTS: In fresh honey, HMF and furosine values ranged from 0.9 to 14.6 mg kg^?1 of product and from 3.06 to 12.06 g kg^?1 of protein, respectively. Heating of honey samples with different pH (3.76 and 5.14) produced slight increases in HMF content and negligible changes were detected in furosine values. The storage of fresh honey for 2 years caused a high increase in the HMF level, reaching values above EU limits. However, furosine showed a different behavior depending on the type of honey sample. Adulteration assays using different syrups produced an increase in HMF and a decrease of furosine values by dilution effect. HMF content of adulterated honey samples with syrup of known origin did not exceed EU limits. CONCLUSION: These results show the influence of long periods of storage or adulteration, using different percentages of corn or invert sugar syrups, on HMF and furosine content of fresh honey. This seems to indicate that the combination of HMF and furosine may be useful for evaluating the quality of fresh honey. Copyright © 2009 Society of Chemical Industry     

Metal content of southern Italy honey of different botanical origins and its correlation with polyphenol content and antioxidant activity

Seventy‐eight samples of southern Italy honey from five different floral origins (chestnut, eucalyptus, citrus, multifloral and sulla) were screened to quantify the polyphenol and metal contents, evaluate the antioxidant activity and determine the correlations between the parameters analysed. The average polyphenol content was 12.06 mg gallic acid equivalent per 100 g honey and 7.92 mg quercetin equivalent per 100 g honey, for total phenolic and flavonoid contents, respectively. The antioxidant activity ranged from 58.40% (eucalyptus honey) to 60.42% (chestnut honey) in the ABTS assay, from 152.65 μm Fe (II) (citrus honey) to 881.34 μm Fe (II) (chestnut honey) in the FRAP assay, and from 54.29% (citrus honey) to 78.73% (chestnut honey) in the DPPH assay. Fe and Zn were the most abundant among the tested metals, while Cd, Co and Mo were those less present. Chestnut honey presented the highest polyphenol content, antioxidant activity and metal content. The correlations between the analysed parameters were statistically significant (P < 0.05). The correlations between metal content and both total phenolic and antioxidant activities were particularly interesting, suggesting a relationship between metal and polyphenol contents in honey.     

Volatile composition and key flavour compounds of spirits from unifloral honeys

The volatile components of industrially produced spirits from three unifloral honeys of campanilla morada (Ipomoea crassicaulis [Benth.] B.L. Robinson), citrus (Citrus spp. L.) and romerillo de costa (Viguiera dentata [Cav.] Spreng.) were studied by combined sensory and instrumental techniques. GC and GC‐MS analyses of the isolated extracts led to the detection of 205 volatile compounds, 150 of them were positively identified. In general, the most abundant classes were saturated alcohols, ethyl esters of saturated fatty acids and terpenes in all samples. Relative aroma intensity of individual volatile components was evaluated by aroma extract dilution analysis of solvent extracts, by gas chromatography‐olfactometry and by odour activity values. Results indicated that 11 compounds: ethanol, ethyl esters of isopentanoic, hexanoic, octanoic and decanoic acids, 2‐phenylethyl acetate, (E)‐β‐damascenone, hotrienol, decanal, methyl anthranilate and ethyl (E)‐cinnamate were the most potent odorants in the honey spirits.     

Effects of conditioning on HMF content in unifloral honeys

HMF (5-hydroxymethylfurfural) is considered an important quality parameter for honey and maximum values are fixed by the European Directive no.110 (2001). In this paper, the HMF levels during the heating process of four unifloral Sicilian honeys (Orange, Eucalyptus, Sulla, Chestnut) were determined; the kinetics of HMF formation were also investigated. The HMF formation was correlated with chemical characteristics (pH, free acids, total acidity and lactones) of the different honeys. The data obtained were statistically elaborated. The results indicate non-equivalence among different honey types with regard to the heating treatment and the importance of reviewing the directive. Thus, the present standards for honey HMF content seem too large in some cases (Chestnut honey) and too restrictive in others (Citrus honey).     

Flavour index and aroma profiles of fresh and processed honeys

This paper discusses the importance of flavour profiling in detecting indicative parameters for quality control of fresh and heated honey. Flavour compounds of six unifloral honeys (Lavandula stoechas, Castanea sativa, Dorycnium pentaphyllum, Rosmarinus officinalis, Eucalyptus sp and Robinia pseudoacacia) were investigated. The aroma extracts were obtained by a two‐step procedure involving (i) preliminary steam distillation under reduced pressure to evaluate the methylpyrazines generated in heated honeys by spectrophotometry (flavour index) and (ii) Likens–Nickerson's simultaneous steam distillation and solvent extraction (SDE) method with added NaCl. A combined total of 64 compounds were detected by gas chromatography/mass spectrometry (GC/MS), 58 of which were identified. Some compounds appeared to be characteristic of the floral source, particularly in Spanish lavender (methylated aliphatic acids and aromatic esters), eucalyptus (2,3‐pentanedione, acetoin, 1‐hexyl alcohol, 2‐acetyl‐5‐methylfuran, furfuryl propionate, 2‐phenylacetaldehyde and nerolidol), chestnut (acetophenone and 2‐aminoacetophenone), rosemary and D pentaphyllum (aromatic acids and esters, 2‐phenylacetaldehyde, farnesol and thymol) honeys. Robinia honey samples were characterised by very low levels of aromatic compounds. Twenty‐six flavour compounds were statistically closely related to the floral origin of the honeys (P ≤ 0.05). The flavour index was evaluated progressively in heated honeys, whereas in fresh honeys it showed a minimal value. © 2003 Society of Chemical Industry     

Characterisation of Slovenian honeys on the basis of sensory and physicochemical analysis with a chemometric approach

The purpose of this study was sensory and physicochemical characterisation of Slovenian honeys with a chemometric approach. Honey samples were obtained from the beekeepers in different natural geographical macroregions of Slovenia. The sensory characteristics of the seven main types of Slovenian honeys are described, together with the physicochemical analyses. The average results of electrical conductivity (0.19–1.61 mS cm^?1), pH (4.01–5.51), free acidity (13.3–30.9 meq kg^?1), proline content (317–558 mg kg^?1), protein content (1.70–3.53 g kg^?1), optical rotation (–19.6 to 12.6), phenolic content (44.9–232.5 mg GAE kg^?1) and antioxidant activity [69.6–456.4 μm Fe(II)] show wide variability among analysed honey types. Statistically significant differences were obtained among different honey types, generally lower values of the analysed parameters were determined in the light honeys, as the acacia, linden and multifloral honeys, while the higher values are characteristic for darker honeys, as the chestnut, fir, spruce and forest honeys. Linear discriminant analysis was performed to classify the honey samples according to their botanical origin and proved that physicochemical parameters analysed can provide enough information for the classification and distinction of acacia, linden, multifloral and chestnut honeys, and the group of honeydew honeys (fir, spruce and forest honeys).     

Stability of anthocyanins in frozen and freeze-dried raspberries during long-term storage: in relation to glass transition

Abstract: Anthocyanins, natural plant pigments in the flavonoid group, are responsible for the red color and some of the nutraceutical benefits of raspberries. This study explores anthocyanin degradation in frozen and freeze‐dried raspberries during storage in relation to glass transition temperatures. Frozen raspberries were stored at ?80, ?35, and ?20 °C, while freeze‐dried raspberries were stored at selected water activity (a_w) values ranging from 0.05 to 0.75 at room temperature (23 °C) for more than a year. The characteristic glass transition temperatures (T′_g) of raspberries with high water content and glass transition temperature (T_g) of raspberries with small water content were determined using a differential scanning calorimeter. The pH differential method was used to determine the quantity of anthocyanins in frozen and freeze‐dried raspberries at selected time intervals. The total anthocyanins in raspberries fluctuated during 378 d of storage at ?20 and ?35, and ?80 °C. Anthocyanin degradation in freeze‐dried raspberries ranged from 27% to 32% and 78% to 89% at a_w values of 0.05 to 0.07 and 0.11 to 0.43, respectively, after 1 y. Anthocyanins were not detectable in freeze‐dried raspberries stored at a_w values of 0.53 to 0.75 after 270 d. First order and Weibull equations were used to fit the anthocyanin degradation in freeze‐dried raspberries. The 1^st‐order rate constant (k) of anthocyanin degradation ranged from 0.003 to 0.023 days^?1 at the selected water activities. Significant anthocyanin degradation occurred in both the glassy and rubbery states of freeze‐dried raspberries during long‐term storage. However, the rate of anthocyanin degradation in freeze‐dried raspberries stored in the glassy state was significantly smaller than the rate of anthocyanin degradation in the rubbery state.     

Unraveling a mechanism of honey antibacterial action: Polyphenol/H2O2-induced oxidative effect on bacterial cell growth and on DNA degradation

Several compounds with antibacterial activities were identified in honey however, a mechanism by which they lead to bacterial growth inhibition and bacterial death remains still unknown. We recently found that honeys possess DNA degrading activity mediated by honey hydrogen peroxide and an unknown honey component(s). Here we provide evidence that active honeys (MIC₉₀ of 6.25–12.5% v/v) possessed significantly higher levels of phenolics (p < 0.02) of higher radical scavenging activities (p < 0.005) than honeys of average activity. Removal of H₂O₂ by catalase eliminated bacteriostatic activities caused by both phenolics and H₂O₂ suggesting that the growth inhibition resulted from the coupling chemistry between these compounds. Both phenolics and H₂O₂ were involved in DNA degradation by honeys. Treatment of plasmid DNA with H₂O₂ alone did not affect the DNA integrity but H₂O₂ removal from honey by catalase prevented DNA degradation. Polyphenols extracted from honeys degraded plasmid DNA in the presence of H₂O₂ and Cu(II) in the Fenton-type reaction. The extent of DNA degradation was inversely related to the polyphenol concentration in this system as well as in honeys. At low content, honey polyphenols exerted pro-oxidant activity damaging to DNA. In conclusion, honey phenolics with pro-oxidant activities were necessary intermediates that conferred oxidative action of H₂O₂. Phenolic/H₂O₂-induced oxidative stress constituted the mechanism of honey bacteriostatic and DNA damaging activities.     

Application of Selected Ion Flow Tube Mass Spectrometry Coupled with Chemometrics to Study the Effect of Location and Botanical Origin on Volatile Profile of Unifloral American Honeys

Abstract: Ten Ohio and Indiana honey samples from star thistle (Centaurea Americana), blueberry (Vaccinium spp.), clover (Trifolium spp.), cranberry (Vaccinium spp.), wildflower, and an unknown source were collected. The headspace of these honeys was analyzed by selected ion flow tube mass spectrometry and soft independent modeling of class analogy (SIMCA). SIMCA was utilized to statistically differentiate between honeys based on their composition. Ohio honeys from star thistle, blueberry, and clover were similar to each other in volatile composition, while Ohio wildflower honey was different. Indiana honeys from star thistle, blueberry, and wildflower were different from each other in volatile composition, while clover and cranberry honeys were similar. Honeys from Ohio and Indiana with the same floral origins were different in volatile composition. Furfural, 1‐octen‐3‐ol, butanoic, and pentanoic acids were the volatiles with the highest discriminating power between types of floral honey. Methanol and ethanol followed by acetic acid were at the highest levels in most honeys, though furfural was at the highest concentration in Indiana blueberry honey, while 1‐octen‐3‐ol was at the highest concentration in Indiana wildflower honey. The highest concentration of volatile compounds was in Indiana wildflower honey followed by Ohio wildflower honey, while the lowest concentration of volatile compounds was observed in Ohio clover honey followed by Indiana clover honey. Practical Application: Using chemometrics, concentrations of volatile compounds in different honeys can be used to determine the influence of botanical and geographical origins on aroma, which is important for the quality of honey. Characterization of volatile compounds can also be a useful tool for assessing honey quality.