Chemical changes in the non-volatile fraction of Brazilian honeys during storage under tropical conditions

The chemical transformations of the non-volatile fraction of Brazilian honeys during storage in tropical condition were monitored. Five systems, namely: 1 – fresh samples; 2 and 3 – samples heated for 3 and 6 months at 35–40 °C; 4 and 5 – samples left under similar conditions to systems 2 and 3, but containing sodium metabisulphite (120 ppm), were tested. The major transformations during storage occurred in the free and lactone acidity, diastase activity and 5-hydroxymethylfurfural (5-HMF) content. Storage modified diastase activity and the content of 5-HMF to an extent that the samples could not remain classified as fresh honey. The action of sodium metabisulphite in the control of the 5-HMF formation seemed to be dependent on other honey characteristics, such as its botanical origin. Sodium metabisulphite also influenced the development of the internal esterification reaction that converts gluconic acid into its corresponding lactone.     

Tracing the geographical origin of honeys based on volatile compounds profiles assessment using pattern recognition techniques

The goal of this study was to examine the possibility of verifying the geographical origin of honeys based on the profiles of volatile compounds. A head-space solid phase microextraction (SPME) combined with comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry (GC × GC–TOFMS) was used to analyze the volatiles in honeys with various geographical and floral origins. Once the analytical data were collected, supervised pattern recognition techniques were applied to construct classification/discrimination rules to predict the origin of samples on the basis of their profiles of volatile compounds. Specifically, linear discriminant analysis (LDA), soft independent modeling of class analogies (SIMCA), discriminant partial least squares (DPLS) and support vector machines (SVM) with the recently proposed Pearson VII universal kernel (PUK) were used in our study to discriminate between Corsican and non-Corsican honeys. Although DPLS and LDA provided models with high sensitivities and specificities, the best performance was achieved by the SVM using PUK. The results of this study demonstrated that GC × GC–TOFMS combined with methods like LDA, DPLS and SVM can be successfully applied to detect mislabeling of Corsican honeys.     

Raw Millefiori honey is packed full of antioxidants

Total polyphenols, flavonoids and antioxidant power of raw honey samples from two of the most common Italian varieties, i.e., Millefiori and Acacia, were evaluated. Phenolic content, expressed as caffeic acid equivalents, ranged from 12.5 to 17.5 mg/100 g and from 3 to 11 mg/100 g in Millefiori and Acacia honeys, respectively. All Millefiori samples exhibited the highest flavonoid concentration being between 1.23 and 2.93 mg catechin equivalents (CE)/100 g honey. Total flavonoids in 100 g Acacia honeys were in the range of 0.45–1.01 mg CE. Acacia honeys had lower total antioxidant power, as assessed by ferric reducing/antioxidant power assay, than Millefiori. The relationship between phenolic content and antioxidant power was discussed. Comparative experimental analysis was performed with an artificial honey and processed honeys. Raw Millefiori honey is rich in both amount and variety of antioxidant substances, and its inclusion in the diet may be recommended to complement other polyphenol sources.     

Nisin–EDTA treatments and modified atmosphere packaging to increase fresh chicken meat shelf-life

The effect of nisin and EDTA treatments on the shelf-life of fresh chicken meat stored under modified atmosphere packaging at 4 °C was evaluated. Chicken meat was subjected to the following antimicrobial treatment combinations: Nisin–EDTA treatments (added post-production to the chicken samples) included: N1 (no nisin–EDTA added; control sample), N2 (500 IU/g; no EDTA added), N3 (1500 IU/g; no EDTA added), N4 (500 IU/g-10 mM EDTA), N5 (1500 IU/g-10 EDTA), N6 (500 IU/g-50 mM EDTA), N7 (1500 IU/g-50 EDTA), N8 (10 mM EDTA; no nisin added), and N9 (50 mM EDTA; no nisin added). N3, N4, N5, N6 and N7 affected populations of mesophilic bacteria, Pseudomonas sp., Brochothrix thermosphacta, lactic acid bacteria, and Enterobacteriaceae. The antimicrobial combination treatments N5, N6 and N7 had a significant effect on the formation of volatile amines, trimethylamine nitrogen (TMA-N) and total volatile basic nitrogen (TVB-N) in chicken meat. The use of MAP in combination with nisin–EDTA antimicrobial treatments resulted in an organoleptic extension of refrigerated, fresh chicken meat by approximately 1–2 days (N2), 3–4 days (N3 and N4), 7–8 days (N5), 9–10 (N7) and by 13–14 days (N6). Chicken was better preserved under treatments N6 and N7, maintaining acceptable odour attributes even up to 24 and 20 days of storage, respectively.     

Effect of ferulic acid on inhibition of polyphenoloxidase and quality changes of Pacific white shrimp (Litopenaeus vannamei) during iced storage

Effects of ferulic acid (FA) on polyphenoloxidase (PPO) and the quality changes of Pacific white shrimp (Litopenaeus vannamei) during iced storage for 10 days were investigated. Both FA and oxygenated FA (OFA) with different concentrations (0.1%, 0.5%, 1% and 2% (w/v)) showed PPO inhibitory activity in a dose dependent manner. FA was generally more effective in PPO inhibition than was OFA. Based on activity staining, white shrimp PPO with an apparent molecular weight of 210 kDa was inhibited by FA. When whole shrimps were treated with FA solution with concentrations of 1% or 2% and stored in ice for up to 10 days, the increase in psychrophilic and mesophilic bacterial count were retarded, in comparison with the control and those treated with 1.25% sodium metabisulphite (SMS). The coincidental lower rates of increase in pH and total volatile base content were obtained. Additionally, shrimps treated with 2% FA possessed the lowest peroxide value and thiobarbituric acid reactive substances (TBARS) value during the storage. After 10 days of storage, shrimps treated with 2% FA had the lower melanosis score and higher score for colour, flavour and overall likeness, compared with the control and SMS treated shrimps (P < 0.05).     

Total and individual carotenoids and phenolic acids content in fresh,refrigerated and processed spinach (Spinacia oleracea L.)

The carotenoid and phenolic acid contents in fresh, stored and processed (blanched, frozen and boiled) spinach were comparatively determined by spectrophotometric and HPLC analyses. The major carotenoids identified after HPLC analysis in saponified samples were lutein (37–53 μg/kg), β-carotene (18–31 μg/kg), violaxanthin (9–23 μg/kg) and neoxanthin (10–22 μg/kg). These carotenoids were all affected by storage and/or heating. The content of carotenoids was best preserved after storage for one day at 4 °C.     

The recognition of high molecular weight melanoidins as the main components responsible for radical-scavenging capacity of unheated and heat-treated Canadian honeys

As the Maillard reaction is known to occur in heat-treated foods, unheated and heated honey samples were subjected to the activity-guided fractionation and size-exclusion chromatography to compare the degree of browning, radical scavenging activity (ORAC) and molecular size of the fractions obtained. Heat-treatment increased browning in the fractions of light- and medium-coloured honeys (p < 0.002), accelerated the formation of high molecular weight melanoidins (85–232 kDa) and significantly increased ORAC values (p < 0.0001). In contrast, melanoidin content and ORAC decreased in the fractions of heat-treated dark buckwheat honey (p < 0.001). Unheated dark honey contained a significantly higher amount of melanoidins than other honeys (p < 0.007). Together, results showed that at low initial concentration of melanoidins, heat-treatment accelerated formation of new melanoidins and increased ORAC, while at high concentration it caused decrease and a reduction of radical scavenging activity. This study emphasises the importance of non-enzymatic browning in the prediction of the antioxidant pool in thermally processed honeys.     

Improvement of the commercial quality of chilled Norway lobster (Nephrops norvegicus) stored in slurry ice: Effects of a preliminary treatment with an antimelanosic agent on enzymatic browning

The use of slurry ice is gaining increasing importance as an advanced method for the hygienic and efficient chilling and sub-zero storage of aquatic food products. In this work, this technology was applied as a novel technique for the chilling and storage of Norway lobster (Nephrops norvegicus) – a crustacean species of high-commercial value – under refrigeration conditions at −1.5 °C. In addition, the effects of a preliminary treatment with 0.5% Na HSO₃ on surface browning were evaluated and compared with the results obtained in control batches not subjected to such treatment. The processing of lobster in slurry ice significantly (p < 0.05) slowed down microbial spoilage, as determined by the counts of aerobes, psychrotrophs, proteolytic bacteria, and lactose-fermenting Enterobacteriaceae, and by the formation of volatile amines. Likewise, the autolytic breakdown mechanisms – as determined by the K value – were also significantly (p < 0.05) inhibited in the slurry ice batch. Remarkably, preliminary treatment with 0.5% sodium metabisulphite permitted better maintenance of the parameters involved in sensory quality – especially as regards the aspect of the carapace – as compared with non-treated batches, and allowed a shelf life of 9 days without surpassing the 150 mg/kg legal limit established for this food additive. On contrast, the non-treated batch stored in slurry ice exhibited a shelf life of 5 days. The combination of technological treatments proposed in this work – preliminary antimelanosic treatment and storage in slurry ice – may be successfully applied to other fresh and frozen shellfish species with a view to extending shelf life and to avoiding the legal and toxicological problems derived from current abuse of such antimelanosic agents to prevent shellfish browning.     

Polyphenol oxidase activity,phenolic acid composition and browning in cashew apple (Anacardium occidentale, L.) after processing

This study describes the extraction and characterisation of cashew apple polyphenol oxidase (PPO) and the effect of wounding on cashew apple phenolic acid composition, PPO activity and fruit browning. Purification factor was 59 at 95% (NH₄)₂SO₄ saturation. For PPO activity, the optimal substrate was catechol and the optimum pH was 6.5. PPO K_m and V_max values were 18.8 mM and 13.6 U min⁻¹ ml⁻¹, respectively. Ascorbic acid, citric acid, sodium sulphite and sodium metabisulphite decreased PPO activity, while sodium chloride increased PPO activity. Wounding at 2 °C and 27 °C for 24 h increased PPO activity but storage at 40 °C reduced PPO activity. Gallic acid, protocatechuic acid and cinnamic acid (free and conjugate) were identified in cashew apple juice. Cutting and subsequent storage at 40 °C hydrolysed cinnamic acid. 5-Hydroxymethylfurfural content in cashew apple juice increased after injury and storage at higher temperatures, indicating non-enzymatic browning.     

Storage-induced chemical changes in active components of honey de-regulate its antibacterial activity

To elucidate reasons for the observed variability in the antibacterial activity of honeys, we analysed a causal relationship between (a) honey floral sources and the activity and (b) the effect of honey storage on stability of compounds conferring this activity. Honeys from diverse floral sources were screened against Escherichia coli (ATCC 14948) and Bacillus subtilis (ATCC 6633) using the broth microdilution method. Among “active” honeys, 37% originated from buckwheat, 18% from clover and 12% from blueberry, indicating that these floral sources produced phytochemical(s) that inhibited bacterial growth. The stability of the putative phytochemical(s) was analysed in “active” honeys (MIC₉₀ 6.25% v/v) by measuring the activity every 3–6 months for a period of 1–3 years. A sharp decline in activity against both bacteria was observed in the first 3–6 months of storage. The decline coincided with major changes in chemical composition of honeys which included a significant change in colour (p < 0.0025), extremely significant change in concentration of UV-absorbing compounds (p < 0. 0001) and appearance of melanoidins. While these changes reduced E. coli sensitivity to honey, it rendered B. subtilis completely insensitive. Thus, the data indicates that the presence of phytochemical(s) conferring the antibacterial activity is sensitive to storage. The de-regulation of the antibacterial activity with the concomitant appearance of melanoidins suggests that the active phytochemical components might be sequestered into melanoidin aggregates, losing their function.     

Honey melanoidins: Analysis of the compositions of the high molecular weight melanoidins exhibiting radical-scavenging activity

Size-exclusion chromatography (SEC) and activity-guided fractionation of honeys allowed the isolation of high molecular weight brown compounds, ranging in size from 66 to 235 kDa that exhibited peroxyl radical-scavenging activity. Their concentrations, antioxidant activity and degree of browning increased after heat-treatment of honeys, suggesting that they represent melanoidins. Chemical analysis of melanoidins demonstrated the presence of proteins, polyphenols and oligosaccharides. Heat-treatment caused an increased incorporation of phenolics into high molecular weight melanoidins and drastically decreased the protein content in these fractions with a concomitant appearance of high molecular weight protein–polyphenol complexes of reduced solubility. LC–ESI–MS demonstrated the presence of oligosaccharide moieties, supporting the postulated origin of melanoidins. The changes in the phenolic content of melanoidins from heated honeys were strongly correlated with their oxygen radical absorbance capacity (ORAC) values (R = 0.75, p < 0.0001), indicating that polyphenols contribute to the antioxidant activity of melanoidins. In summary, honey melanoidins are multi-component polymers consisting of protein–polyphenol–oligosaccharide complexes. A direct interaction between polyphenols and melanoidins resulted in a loss or gain of function for melanoidin antioxidant activity.     

Evaluation of the phenolic content,antioxidant activity and colour of Slovenian honey

Honey samples from the seven most common honey types in Slovenia were screened for total phenolic content by the modified Folin–Ciocalteu method, for potential antioxidant activity using the ferric reducing antioxidant power (FRAP) assay and by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method for antiradical activity. In addition the colour characteristics of honey samples were analysed. The results of the study showed that total phenolic content, antioxidant activity and colour parameters differ widely among different honey types. Phenolic content expressed as gallic acid equivalent ranged from 44.8 mg/kg in acacia honey to 241.4 mg/kg in fir honey. Antioxidant activity was the lowest in the brightest acacia and lime honeys and the highest in darker honeys, namely fir, spruce and forest. The colour of the Slovenian honeys, analysed in this study was very variable and ranged from pale yellow to dark brown. Correlations between the parameters analysed were found to be statistically significant (p < 0.05).     

Assessment of physical and mechanical properties of sodium caseinate and stearic acid based film-forming emulsions and edible films

Edible films were prepared using sodium caseinate (6–8 g/100 g) and stearic acid (0–2 g/100 g). Effects of the ratio of stearic acid and sodium caseinate to water on the water vapor permeability (WVP) and mechanical properties of the prepared films were evaluated. Film-forming emulsions were also tested for rheological properties and surface tension. Changes in the ratios of sodium caseinate and stearic acid to water had significant effects on WVP (p < 0.05) and surface tension (p < 0.01). Higher values of consistency coefficient and elastic modulus were obtained in the presence of higher stearic acid. In addition, increase in stearic acid content increased the rate of water loss and gain of elastic modulus at the early stage of drying and resulted in production of less flexible film. The resultant edible film prepared with 6 g/100 g sodium caseinate and 2 g/100 g stearic acid showed the lowest WVP of 1.368 (g mm/m² h kPa).     

Possible role of volatile amines as quality-indicating metabolites in modified atmosphere-packaged chicken fillets: Correlation with microbiological and sensory attributes

The present work evaluated the possible role of volatile amines as indicator(s) of poultry meat spoilage. Fresh chicken meat (breast fillet) was packaged in four different atmospheres: air (A), vacuum (VP) and two modified atmospheres (MAs), namely M1, 30%/65%/5% (CO₂/N₂/O₂) and M2, 65%/30%/5% (CO₂/N₂/O₂). All chicken samples were kept under refrigeration (4 ± 0.5 °C) for a period of 15 days. Of the four treatments, the VP and M1 and M2 gas mixtures were the most effective for delaying the development of aerobic spoilage microbial flora. Pseudomonas spp. in chicken samples stored under M2 gas mixture and VP were significantly lower than all the other samples after 15 days of storage. Of the remaining bacterial species examined, lactic acid bacteria (LAB), Brochothrix thermosphacta, were dominant in the microbial association of both aerobically- and MA-packaged chicken, while yeasts contributed to a much lesser extent in the final microbial flora of chicken meat. On the basis of microbiological data (TVC), shelf-life extensions of 2, 4 and 9–10 days were achieved by VP and M1 and M2 gas mixtures. Results of the present work showed that the limit of sensory acceptability (a score of 6) was reached for the aerobically, vacuum-packaged and M1 gas mixture chicken samples approximately on days 6–7 and 9–10, respectively. Based on sensory (taste) analysis and with regard to chicken spoilage and freshness, TMA-N and TVB-N limit values of acceptability, namely 10.0 mg N/100 g and 40 mg N/100 g for chicken samples stored in air, may be proposed as the upper limit values for spoilage initiation of fresh chicken meat stored aerobically. Interestingly, the M2 gas mixture sample did not reach these limit values throughout the 15 day storage period. The formation of volatile amines during chill storage of chicken meat, under the packaging conditions examined in the present study, seemed to be in good agreement with the increase in microbiological count (TVC) and sensory taste score except for the M2 gas mixture.     

Composition of volatile compounds of honey of various floral origin and beebread collected in Lithuania

Honey is collected from various flowering plants and its composition, particularly volatile flavour compounds to some extent depends on the nectar source. Therefore, some volatile constituents may be indicators of honey origin. In this study the volatile profiles of 15 honey samples of different botanical origin and one beebread sample are characterised. Volatiles were collected by means of SPME and analysed by GC/MS. Botanical source of honey samples was established by the melissopalynological method: 11 of analysed samples were unifloral rape honeys, 1 clover, 1 caraway and 2 polyfloral. In total 93 compounds in honey and 32 in beebread were identified. They involve different classes of chemical compounds, including alcohols, ketones, aldehydes, acids, terpenes, hydrocarbons, benzene, and furan derivatives. Benzaldehyde and benzenacetaldehyde were the only compounds found in all 15 honey sample. Dimethyl sulphide, pentanenitrile, benzylnitrile were identified in 14 honeys; isobutane, octanoic and nonanoic acids in 13 samples; furfural, linalool and nonanal in 12 samples; octanal, lilac aldehyde C, hotrienol and decanal in 11 samples and finally 2-methylbutanenitrile in 10 honey volatile fractions. Remarkable variations were observed in the composition of volatiles in honey from different sources. In addition, volatile profiles of honey samples were analysed after 3 months of storage and it was found that the amount of headspace volatiles in the majority of samples decreased.     

Organic acids,antioxidant capacity,phenolic content and lipid characterisation of Georgia-grown underutilized fruit crops

Four underutilized Georgia-grown fruit crops, namely loquat (Eriobotrya japonica), mayhaw (Crataegus sp.), fig (Ficus carica), and pawpaw (Asimina triloba), and their leaves were analysed for total polyphenols by Folin–Ciocalteau method, and antioxidant capacity by ferric-reducing antioxidant power (FRAP) and Trolox-equivalent antioxidant capacity (TEAC) assays. Organic acids and phenolic compounds were identified by RP-HPLC. For lipid profile, fruits were separated into two fractions – seed and fruit (i.e., without seed); lipid was extracted using the Folch method and analysed for fatty acids, phytosterols, tocopherols, and phospholipids. The major organic acid identified in all samples was malic acid (177–1918 mg/100 g FW). The predominant phenolic acids in all the fruits were gallic (1.5–6.4 mg/100 g FW) and ellagic (0.2–33.8 mg/100 g FW), and the most abundant flavonoid was catechin (12.2–37.8 mg/100 g FW). Total lipid content varied from 0.1% in mayhaw fruit to 21.5% in pawpaw seed. Linoleic acid was the predominant fatty acid in all of the samples (28.2–55.7%).     

Concentrations of medium-chain 2- and 3-hydroxy fatty acids in foodstuffs

Medium chain 2-hydroxy fatty acids and 3-hydroxy fatty acids (2-OH-FAs and 3-OH-FAs with 8–20 carbons) are widespread in nature but little was known about the quantities of these minor fatty acids in food. For this reason, the concentrations and composition of 2- and 3-OH-FAs in several foodstuffs (milk and dairy products, animal brains, suet, vegetable oils) and two non-food samples (wool wax and vernix caseosa) were determined by gas chromatography with electron-capture negative ion mass spectrometry in the selected ion monitoring mode. Lipids were isolated from samples by accelerated solvent extraction. After transesterification the obtained fatty acid methyl esters (FAMEs) were separated into OH-FAMEs and non-OH-FAMEs. The OH-FAMEs were converted into pentafluorobenzoyl (PFBO) derivatives (PFBO-O-FAMEs). 2- and 3-OH-FAs were detected in all samples analysed. Chain-length ranged from 8–20 carbons. In general, 3-OH-FAs dominated in milk and dairy products (maximum concentration 17.7 mg/100 g l.w. in goat milk) whereas 2-OH-FAs were more abundant in animal brains and suet. The lowest concentrations were determined in vegetable oils (0.21–2.42 mg/100 g l.w.) whereas the highest concentrations of 2-OH-FAs were determined in wool wax (1180 mg/100 g l.w.) and vernix caseosa (34.5 mg/100 g l.w.).     

Inhibition of melanosis formation in Pacific white shrimp by the extract of lead (Leucaena leucocephala) seed

Lead (Leucaena leucocephala) seed extract was prepared using distilled water as a medium. An extraction yield of 26.16 g/100 g of seed was obtained after extraction at room temperature for 12 h. Total phenolic and mimosine contents in the lead seed extract powder (LSEP) were 17.4 g GAE/100 g and 8.8 g/100 g, respectively. LSEP at different concentrations (0.05%, 0.1%, 0.25%, 0.5%, and 1%, w/v) showed inhibitory activity towards polyphenoloxidase (PPO) of Pacific white shrimp in a dose dependent manner. When the whole Pacific white shrimp were treated with 0.25% and 0.5% (w/v) LSEP, the shrimp treated with 0.5% LSEP had the lower melanosis score throughout the storage of 12 days and showed a higher score for colour and odour, as well as overall likeness, compared with the control (without treatment) and 1.25% sodium metabisulphite treated samples at day 12 (P < 0.05). Meat of shrimps treated with LSEP at both levels had the increase in mimosine content up to 8 days, suggesting the migration of mimosine into shrimp muscle during extended storage. Therefore, 0.5% LSEP can be used as a novel melanosis inhibitor for Pacific white shrimp.     

Fatty acids and cholesterol oxidation in salted and dried shrimp

The aim of this work was to evaluate the occurrence of cholesterol oxidation products and to analyze the lipidic profile in salted–dried shrimp. Fifty samples of salted–dried shrimp were evaluated, and the cholesterol oxides (7β-OH, 7α-OH, 7-Keto and 25-OH) were quantified by high-performance liquid chromatography. The cholesterol oxides: 7β-OH (34.63–72.56 μg/g), 7α-OH (5.02–12.12μg/g), 7-Keto (7.44–32.68 μg/g) and 25-OH (2.37–22.88 μg/g) were determined in all samples analyzed. Regarding to the total cholesterol content and the average thiobarbituric acid reactive substances (TBARS) content, the results ranged from 73.88 to 247.69 mg/100 g, and 0.02 to 1.30 mgMA/kg, respectively. The fatty acids profile was: 27.48% saturated, 43.90% monounsaturated and 28.61% polyunsaturated. The presence of cholesterol oxidation products and the values of TBARS indicate the degree of oxidation of this product, which was probably initiated by inadequate conditions of processing and storage.     

Flavonoid pattern of sage (Salvia officinalis L.) unifloral honey

The aim of the present paper was to determine the flavonoids in monofloral sage (Salvia officinalis L.) honey which is characteristic and specific for the area of Croatian coast and islands. For that purpose 38 sage honey samples from two production seasons were analysed. After specific pollen content determination, and analyses of selected physicochemical parameters which confirmed that samples are in compliance with national and international regulations and can be regarded as unifloral sage honeys, flavonoid fraction was isolated and analysed using RP-HPLC/DAD method. The HPLC analysis showed that all examined sage honey samples contain quercetin (3,3′,4′,5,7-pentahydroxyflavone), luteolin (3′,4′,5,7-tetrahydroxyflavone), kaempferol (3,4′,5,7-tetrahydroxyflavone), apigenin (4′,5,7-trihydroxyflavone), chrysin (5,7-dihydroxyflavone) and galangin (3,5,7-trihydroxyflavone), as well as p-coumaric (trans-4-hydroxycinnamic acid) and caffeic acid (3,4-dihydroxycinnamic acid). Total amount of identified flavonoids varied from 109.4 μg/100 g of honey to 589.9 μg/100 g of honey, with the average of 288.5 μg/100 g of honey. All analysed honey samples showed common and specific flavonoid profile which could be the basis for differentiating sage from other monofloral honeys.