Chemical and molecular dynamics analysis of crystallization properties of honey

Honey crystallization is a natural phenomenon commonly found in honey products. To better understand the characteristics of honey crystallization, the concentrations of the four major compositions (namely glucose, fructose, sucrose, and maltose) were determined in the liquid, crystal, and mixed forms of honey. Our study revealed the contents of glucose and fructose in three forms varied significantly (p < 0.05). Molecular dynamics simulation was performed to investigate honey crystallization process. All the simulation systems had a tendency to form a cubic crystal. When the ratio of glucose/fructose is 2.5:1, which is the same as the crystal found in honey, a shorter time and lower root-mean-square deviation were found compared to the ratios at 2:1, 1:1, and pure glucose. It infers that glucose/fructose at 2.5:1 is the most stable honey crystallization, relatively. The morphology of crystals found in honey, which was observed under an environment scanning electron microscope, looked like sticky balls with indistinct edges, different from that plates or granule of glucose crystal with flattened edges. These results indicated that the relative content of glucose and fructose had a significant effect on the crystallization of honey. The glucose/fructose at 2.5:1 probably is the critical ratio of honey crystallization, at which honey can form stable crystalline deposit speedily.     

Vitamin C and sugar levels as simple markers for discriminating Spanish honey sources

In this work, 7 Spanish honeys with different botanical origins were studied. The honey origins were rosemary, chestnut, lavender, echium, thyme, multifloral, and honeydew. The chemical compounds determined were ascorbic acid (vitamin C), hydroxymethylfurfural, and major sugar contents (glucose and fructose). The physicochemical parameters, pH, conductivity, moisture, free acidity, and color, were also measured. Vitamin C is an important antioxidant in food, and the possibility to use it as discriminate parameter among different honeys was studied. The determination of vitamin C in honey samples was carried out by 2 different methods, volumetric and chromatographic comparing the results by both statistically. Vitamin C content was higher in thyme honeys than in the other types; however a wide dispersion in the values was found. Through a linear discriminant analysis (LDA), conductivity, glucose, fructose, and vitamin C content were the most important discriminant parameters. PRACTICAL APPLICATION: Vitamin C content in different honey sources has been determined by a simple and rapid chromatographic method (less than 3 min) in honeys from 6 botanical origins. The results together with glucose and fructose content and some physicochemical parameters have been studied in order to discriminate the botanical origin of honeys and in the future certified their quality. A statistical LDA was applied to the data, and differentiation of honey sources was possible with very good agreement. The vitamin C content found in thymus honeys was significantly higher than in other types. This fact makes vitamin C a special marker for thymus honeys that have a higher antioxidant effect than the others giving it special properties. The identification of honey sources is essential for beekeepers in order to certify honeys for consumers.     

Carbohydrate composition and electrical conductivity of different origin honeys from Lithuania

The main task of this study was to characterize Lithuanian honeys obtained from various sources by their carbohydrate composition and electrical conductivity and to determine if there is any dependence between these characteristics and pollen content. Twenty six samples of honey collected in Lithuania in 2006 during flowering season were analyzed by gas chromatography. Botanical source of honey samples was established by the melissopalynological method: 15 of analyzed samples were unifloral rape (winter and spring), 7 willow and 4 polyfloral honeys. Fructose, glucose, sucrose, maltose, isomaltose, turanose, trehalose, palatinose, cellobiose, raffinose and panose were identified and quantified in all samples. Glucose was predominant in 22 out of 26 samples. The mean values of fructose and glucose varied from 329.2 to 400.0 and from 346.0 to 426.3 mg/g honey, respectively. The amount of sucrose was 0.7-2.5 mg/g. Some correlations between sugar concentration and the content of pollen were established. The ratios of fructose/glucose, maltose/isomaltose, maltose/turanose, sucrose/turanose, which may be used as indicators for honey authenticity, were calculated. In addition, electrical conductivity was measured and it was found that it varied from 0.27 to 0.89 mS/cm. Unifloral rape honeys had the lowest electrical conductivity. Data obtained was thoroughly compared with previously published results and it was found that the characteristics of Lithuanian honeys in most cases meet international requirements for natural honey. However, the information on honey sugar composition and electrical conductivity was not sufficient for the reliable determination of the botanical origin of honey.     

Influence of Different Unifloral Honeys on Heterocyclic Aromatic Amine Formation and Overall Mutagenicity in Fried Ground-beef Patties

ABSTRACT The effects of different unifloral honeys (buckwheat, clover, and sage), carbohydrates (fructose, glucose, and sucrose), and antioxidants (vitamin E, BHT), and Trolox® (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) on heterocyclic aromatic amine (HAA) formation and overall mutagenicity in fried ground patties were evaluated. The inhibition of total HAA formation was achieved with buckwheat (55%), clover (52%), and sage (51%); and they also reduced overall mutagenicity 36, 31, and 26%, respectively. The addition of fructose, glucose, or fructose and glucose together, at levels comparable to their occurrence in honey, reduced (p < 0.05) mutagenicity and HAA formation by amounts comparable to that found with honey.     

Determination of non-volatile compounds of different botanical origin Brazilian honeys

The levels of water, HMF, free proline, total acidity, diastase activity, fructose and glucose in 74 different floral type honeys from four Brazilian regions are reported. The majority of the samples showed adequate water and HMF contents indicating the use of good practices by beekeepers in Brazil. Large variation in the contents of proline (389–520 mg/kg) was found in the honeys from the four regions studied. These findings are presumably due to the more intensive labor of the bees on the collected nectar by adding gland secretions. The high amount of diastase (40–120°G) found in some honeys from the northeast region could be due to the visit of the bees to Manhiot sp. (cassava) flour factories to collect starch-rich food, consequently increasing the diastase activity in the honey. Xerotolerant yeasts may be responsible for higher total acidity and ratios below 1 for fructose/glucose found in some samples. The majority of the analysed honeys (82%) were within the limits of the European Codex Honey Standards and Brazilian legal regulations.     

Aroma profile and physico‐chemical properties of artisanal honey from Tabasco,Mexico

The aim of this work was to characterise, physico‐chemically, sugar composition and aroma profile of ten honey samples collected by small producers during two seasons and produced in different agricultural ecosystems from Tabasco State (Mexico). The mean values obtained for the physico‐chemical parameters were: pH 3.63; 19.25% moisture; 78.8°Brix sugar; 0.64% ash; 0.25 mS cm^?1 electrical conductivity and 0.58 water activity. Based on colour parameters, Tabasco honeys can be placed in the group of dark honeys. In the analysed samples; fructose was the major sugar (39.45%), followed by glucose (35.74%) while sucrose only represented 2.93%. The volatile profiles of the ten honey samples were obtained by gas chromatography‐mass spectrometry. All the honeys were characterised by their high contents of benzene and furan‐related compounds. As a result, artisanal honeys from Tabasco (Mexico) can be considered to present a good level of quality.     

Physical Properties of Co-crystalline Sugar and Honey

ABSTRACT: Clear and partially granulated honeys, were co-crystallized with sucrose in 3 proportions, 90:10, 85:15, 80:20, respectively. The products obtained by centrifugal filtration from both honeys were found by high-performance liquid chromatography to contain between 1% and 2% of honey. The moisture content of the products increased with the amount of honey added, whereas their powder flowability decreased. Differential scanning calorimetry showed a decrease in the overall crystallinity of the solidified products with an increase in the amount of honey added. Neither crystalline fructose nor glucose was detected by powder X-ray diffraction in any of the co-crystalline products. Glucose monohydrate crystallized on storage in some co-crystalline materials prepared from granulated honey.     

Evolution of fructose and glucose in honey over one year: influence of induced granulation

Evolution of fructose and glucose over 1 year has been evaluated in 30 honey samples from Burgos (N. Spain). The influence of the induced granulation process in this evolution was also determined. Each sample was divided into two aliquots of 500 g and aseptically bottled. One aliquot was directly stored and the second induced to crystallise by seeding with 10% of finely crystallised honey. Analyses of moisture content, pH, fructose and glucose were carried out over 1 year, once each 4 months. Both, fructose and glucose increased in most samples. Induced-crystallised samples did not show any significant differences in the evolution of the two sugars in comparsion with directly stored samples. Linear correlations were found, for both fructose and glucose, between samples directly stored and honeys in which granulation was induced. These results are clearly different from those reported in previous papers where decrease of monosaccharides below their original values was described. pH of honey might promote reversion of monosaccharides and the formation of disaccharides and trisaccharides. This investigation has demonstrated the possibility of formation of monosaccharides, by the hydrolysis of higher sugars, as a process predominant over the reversion. No statistical relationship was found between pH of honey samples and their fructose and glucose content evolution.     

Rapid analysis of sugars in honey by processing Raman spectrum using chemometric methods and artificial neural networks

The aim of this study was to quantify glucose, fructose, sucrose and maltose contents of honey samples using Raman spectroscopy as a rapid method. By performing a single measurement, quantifications of sugar contents have been said to be unaffordable according to the molecular similarities between sugar molecules in honey matrix. This bottleneck was overcome by coupling Raman spectroscopy with chemometric methods (principal component analysis (PCA) and partial least squares (PLS)) and an artificial neural network (ANN). Model solutions of four sugars were processed with PCA and significant separation was observed. This operation, done with the spectral features by using PLS and ANN methods, led to the discriminant analysis of sugar contents. Models/trained networks were created using a calibration data set and evaluated using a validation data set. The correlation coefficient values between actual and predicted values of glucose, fructose, sucrose and maltose were determined as 0.964, 0.965, 0.968 and 0.949 for PLS and 0.965, 0.965, 0.978 and 0.956 for ANN, respectively. The requirement of rapid analysis of sugar contents of commercial honeys has been met by the data processed within this article.     

Differentiation of blossom and honeydew honeys using multivariate analysis on the physicochemical parameters and sugar composition

Multivariate analysis was applied on physicochemical parameters (moisture, water activity, electric conductivity, colour, hydroxymethyl furfural, acidity, pH, proline, diastase and invertase), sugar composition (fructose, glucose, sucrose, maltose, isomaltose, trehalose, turanose and melezitose) and palinological parameters determined in blossom and suspected honeydew honeys in order to differentiate them. The majority of the physicochemical, sugar composition and palinological parameters evaluated presented significant differences in the mean values between the suspected honeydew and blossom honeys, with the exception of moisture, water activity, diastase, fructose and maltose. Blossom honey samples tend to differentiate from the suspected honeydew honeys after applying factor analysis on the physicochemical parameters and sugar composition. Stepwise linear discriminant analysis allows the correct classification of all blossom honeys, and only one honeydew honey was erroneously included as blossom honey. So, the use of multivariate analysis on physicochemical parameters and sugar composition can be a useful tool to differentiate these types of honeys.     

Sugar profile and physicochemical properties of Ethiopian monofloral honey

Three hundred twenty honey samples were collected from Ethiopia, and categorized into nine monofloral honeys using melissopalynology. Theses monofloral honeys were investigated for their sugar profile, physicochemical properties, and quality of honeys. Sugars (fructose, glucose, sucrose, maltose, turanose, and isomaltose) were identified using high-performance liquid chromatography and physicochemical properties (moisture, water activity, pH, acidity, ash, minerals, conductivity, colour) using the Harmonized Method of the International Honey Commission. Moisture and water activity showed a strong regression coefficient (r² = 0.92). Moisture content alone did not sufficiently describe the quality of honey during storage and processing.     

Effect of honey high-temperature short-time heating on parameters related to quality, crystallisation phenomena and fungal inhibition

The effect of a unique high-temperature short-time heating treatment on natural honeys to produce both a fungi inhibition and a crystallisation delay simultaneously was evaluated. As responses to treatments, hydroxymethylfurfural content, diastase activity, fungi and yeast development, and crystallisation starting time were measured. Besides, moisture, fructose, glucose and acidity were determined.Heating can produce a decrease in honey quality, which is made evident by a simultaneous reduction in the diastase activity, referred to as diastase number, and an increase in the hydroxymethylfurfural content. Considering the minimum admissible diastase number and the maximum admissible hydroxymethylfurfural content values, according to honey present regulations, several thermal treatment tests were performed on selected honey samples without exceeding those values. Tested conditions were: heating between 140°C and 80°C at times between 60 to 15 s and 30 to 10 s during both transient and isothermal heating stages, respectively. In all tests the diastase number was the most heat-sensitive parameter. The boundary thermal treatment, assumed as the most severe conditions able to produce a permissible quality loss, was found to be 140°C, during 15 s in the transient stage and 30 s in the isothermal stage. Beyond those values, an unacceptable diastase number reduction was produced. On the other hand, it was found that a 80°C heating, during 60 and 30 s in transient and in isothermal stages, respectively, destroyed all microorganisms responsible for quality damage without spoiling honey. This was considered to be the mild or more suitable treatment condition. Twenty-six naturally contaminated honey samples heated at such conditions showed a null capacity to develop fungi and yeast. Besides, according to the storing temperature, and moisture and glucose contents, a crystallisation beginning delay between 4 and 9 weeks was obtained on four selected honeys, according to the thermal treatment applied. Several parameters previously proposed that characterise honey crystallisation tendency as fructose/glucose, glucose/moisture and (glucose-moisture)/fructose ratios were related to the time in which crystallisation starts. A new parameter, (fructose/glucose)·moisture was proposed.     

Effect of Adulteration versus Storage on Volatiles in Unifloral Honeys from Different Floral Sources and Locations

High fructose corn syrup (HFCS) was added at 5% to 40% to Indiana wildflower honey and added at 40% to Ohio and Indiana honeys from blueberry, star thistle, clover and wildflower, and an unknown source to simulate honey adulteration. Unadulterated honeys were also stored at 37 ºC from 1 to 6 mo. The volatile composition was measured by Selected Ion Flow Tube Mass Spectrometry (SIFT‐MS). Most volatiles decreased in concentration with both increasing HFCS and storage time. Furfural significantly increased in concentration in all adulterated honeys and 1,3‐butanediol, acetonitrile, and heptane in some adulterated honeys. During storage, the volatiles that increased were maltol, furfural, 5‐methylfurfural, and 5‐hydroxymethyl furfural in all honeys and also acetic acid and 1‐octen‐3‐ol levels in some honeys. Soft independent modeling by class analogy (SIMCA) was used to differentiate the volatile profiles of adulterated honeys from fresh and stored honeys. The volatile profiles of honeys in accelerated storage for up to 4 mo and the honeys adulterated with 40% HFCS were significantly different. Acetic acid had the most discriminating power in Ohio star thistle and blueberry honeys and unknown honey while furfural had the greatest discriminating power in Indiana blueberry, star thistle, and clover honeys. Adulteration and storage of honey both reduced the volatile levels, but since they changed the volatile composition of the fresh honey differently, SIMCA was able to differentiate adulteration from storage. Practical Application: Analysis of adulterated and stored honeys determined that both decrease volatile levels, and no clear indicator volatiles were found. However, SIMCA can be used to distinguish the volatile profiles of fresh or stored honeys, from adulterated honeys.     

Honey Adulteration Detection Using Raman Spectroscopy

In this study, the Raman spectroscopy was used to detect honey adulterated with fructose (F), glucose (G), inverted sugar (IS), hydrolyzed inulin syrup (IN), and malt must (M). Thus, 56 samples of authentic honeys (acacia, sunflower, tilia, polyfloral, and honeydew) and 900 adulterated samples (with 5, 10, 20, 30, 40, and 50% fructose, glucose, inverted sugar, malt must, and hydrolyzed inulin syrup) were analyzed. The classification of honey authenticity has been made using the partial least square linear discriminant analysis (PLS-LDA), and a total accuracy of 96.54% (authentic honey vs. adulterated honey) was observed, while in the case of adulterated honey, a total accuracy of 90.00% was observed, respectively. The determination of the adulterant agent concentration has been made using partial least squares regression (PLSR) and principal component regression (PCR) methods. The proposed method can be considered easy and rapid for honey adulteration detection to provide continuous in-line information.     

The impact of ultrasound decrystallization on enzymatic,antioxidant and antibacterial properties of honey

The aim of the research was to assess the impact of liquefying honey by ultrasound processing on its biological activity. Four solid honey samples of different botanical origin were decrystallized by using the ultrasound (sonic bath, 40 kHz, 800 W) and conventional thermal processing (water bath, 40 °C). The biological properties of liquefied and control solid honeys were compared. The ultrasound processing significantly shortened the time of honey liquefaction (from 24 h to 15 min) and delayed of the re-crystallization process compared to conventional method (from 1 to 4 months). Diastase number was comparable in ultrasound and unprocessed honeys, but decreased in conventionally liquefied samples (by 8 to 20.6%) whereas the activity of α-mannosidase and β-galactosidase was insignificantly affected only. Growth inhibition of S. aureus and E.coli by 12.5% water solution of control honey samples was variety-dependent, the lowest for rape (12 and 19%) and the highest for honeydew (32 and 38%, respectively). No significant changes were observed after ultrasound treatment. The antioxidant properties of ultrasound processed honeys were insignificantly reduced up to 8%, while total polyphenols content increased by 3–6% in 75% of tested samples. The hydroxymethylfurfural (HMF) content significantly increased after both treatments (from 31 to 40%), however without exceeding the allowable limit.Industrial relevanceHoney crystallization during storage is natural but undesirable serious problem in processing and marketing of the honey. The ultrasound processing shortened the processing time (saved costs), limited the honey biological activity deterioration, and delayed the re-crystallization of processed honeys. Ultrasound processing is a beneficial non-thermal alternative for conventional liquefying of honey by heating.     

Chemical Composition and Temperature Influence on the Rheological Behaviour of Honeys

The purpose of this work was to examine the viscoelastic properties of Spanish honeys with various sugar contents [fructose (32–42 g/100 g honey), glucose (24–35 g/100 g honey), sucrose (0.0–3.4 g/100 g honey)]; concentrations (79–83 °Brix), and moisture levels (16–19 g/100 g honey) at different temperatures (5, 10, 15, 20, 25, 30, and 40°C). Honey showed Newtonian behaviour, presenting a highly viscous part (loss modulus was much greater than the elastic modulus). The loss modulus (G″) and viscosity increased with moisture content and a decrease with temperature. Exponential and power law models were applied to fit loss modulus and viscosity data. Polynomial models were proposed to describe the combined effect of temperature, fructose, glucose, sucrose content, other sugars, non-sugar substance, and moisture content.     

Rheology and crystallization kinetics of honey: Present status

Abstract

Crystallization property of honey is a matter of interest for beekeepers, honey handlers and processors. Honey is basically a sugar syrup with about 16‐18% moisture content (wet basis). The composition and rheology are some of the important parameters that influence the crystallization behavior of honey. At present, most of the crystallization prediction methods are based on the composition considering honey as a glucose‐fructose‐water system. This review paper presents current methods used to predict honey crystallization and puts forward a hypothesis based on viscosity and glass transition temperature.     

Physicochemical and melissopalynological characterization of Estonian summer honeys

The samples of 14 honeys, retained from Estonian beekeepers, were analyzed for parameters such as pH, moisture content, free acidity, electrical conductivity, diastase activity, hydroxymethylfurfural (HMF) content and mineral content, including sodium (Na), potassium (K), magnesium (Mg) and calcium (Ca). Fructose, glucose and disaccharide content were also identified and fructose/glucose ratio was calculated. In addition melissopalynological analyses were carried out for characterization of honeys. The mean values of analyzed honeys were: pH 3.8; moisture 17.3%; free acidity 20.4 mmol/kg; electrical conductivity 0.2 mS/cm; diastase activity 23.1 DN and HMF was below 3.8 mg/kg. Within the mineral content, potassium was quantitatively the most important mineral in the range of 125.79 to 1381.53 mg/kg followed by calcium of 20.37-63.65 mg/kg, magnesium 5.53-25.49 mg/kg and sodium 4.77- 19.44 mg/kg. The predominant sugar in honey samples was fructose having the mean value of 35.91 g/100 g followed by glucose 35.00 g/100 g. The disaccharide average content was 6.00 g/100 g. The melissopalynological analyzes showed that the most dominant pollens in honey samples were cruciferous (Cruciferae)-mainly rape (Brassica napus); rosacean (Rosaceae)-mainly raspberry (Rubus idaeus); white clover (Trifolium repens); sweet clover (Melilotus officinalis) and willow (Salix). The results of honey pollen profile analysis and calculated fructose/glucose ratios (0.89-1.17) both indicated to flower honeys. All of the analyzed honeys were found to meet European Legislation (EC Directive 2001/110) for all parameters.     

Pyrrolizidine alkaloids in honey and bee pollen

A total of 3917 honey samples and 119 'bee pollen' samples (pollen collected by honeybees) were analysed for pyrrolizidine alkaloids (PAs). Some 0.05 M sulphuric acid was used for extraction followed by a clean-up step by means of solid-phase extraction. Separation and detection was achieved by target analysis using an LC-MS/MS system. PAs were found in 66% of the raw honeys (bulk honey not yet packaged in containers for sale in retail outlets) and in 94% of honeys available in supermarkets (retail honey). A total of 60% of the bee pollen samples were PA positive. The PA pattern was used to identify the potential origin of the PAs in honey, which was verified for the genus Echium by relative pollen analysis. The results give an estimate of the impact of PA-containing plants belonging to the genera Echium, Senecio and, to a certain extent, Eupatorium on PA levels in honey and can serve as a decision basis for beekeepers in order to find the most suitable location for the production of honey and bee pollen low in PAs.     

Color changes during storage of honeys in relation to their composition and initial color

The causes of darkening in honey have been attributed to Maillard reaction, fructose caramelization and reactions of polyphenols, however, no systematic studies exist on this subject. The influence of composition and initial color on the rate of darkening of several Argentine honeys submitted to storage at 37°C during 90 days was evaluated through spectrocolorimetric measurements. The most suitable color functions to evaluate darkening of honeys [lightness (L_ab*), browning index (BI), metric chroma (C_ab*), metric hue (H_ab*) and 1/Z] increased linearly as storage time increased, after an initial induction period of very low browning development. The slope of the linear browning development zone with time was an index of browning rate, and it was analyzed in relation to the initial color and the composition of honeys (moisture content, total nitrogen, total lipids and polyunsaturated fatty acids, fructose and glucose content). Of the analyzed variables, the initial color was the parameter which better defined the rate of darkening of honeys.