The outcome of boosting mitochondrial activity in alcohol-associated liver disease is organ-dependent.

BACKGROUND AND AIMS: Alcohol-associated liver disease (ALD) accounts for 70% of liver-related deaths in Europe, with no effective approved therapies. Although mitochondrial dysfunction is one of the earliest manifestations of alcohol-induced injury, restoring mitochondrial activity remains a problematic strategy due to oxidative stress. Here, we identify methylation-controlled J protein (MCJ) as a mediator for ALD progression and hypothesize that targeting MCJ may help in recovering mitochondrial fitness without collateral oxidative damage. APPROACH AND RESULTS: C57BL/6 mice [wild-type (Wt)] Mcj knockout and Mcj liver-specific silencing (MCJ-LSS) underwent the NIAAA dietary protocol (Lieber-DeCarli diet containing 5% (vol/vol) ethanol for 10 days, plus a single binge ethanol feeding at day 11). To evaluate the impact of a restored mitochondrial activity in ALD, the liver, gut, and pancreas were characterized, focusing on lipid metabolism, glucose homeostasis, intestinal permeability, and microbiota composition. MCJ, a protein acting as an endogenous negative regulator of mitochondrial respiration, is downregulated in the early stages of ALD and increases with the severity of the disease. Whole-body deficiency of MCJ is detrimental during ALD because it exacerbates the systemic effects of alcohol abuse through altered intestinal permeability, increased endotoxemia, and dysregulation of pancreatic function, which overall worsens liver injury. On the other hand, liver-specific Mcj silencing prevents main ALD hallmarks, that is, mitochondrial dysfunction, steatosis, inflammation, and oxidative stress, as it restores the NAD + /NADH ratio and SIRT1 function, hence preventing de novo lipogenesis and improving lipid oxidation. CONCLUSIONS: Improving mitochondrial respiration by liver-specific Mcj silencing might become a novel therapeutic approach for treating ALD.

Preventing Mislabeling: A Comparative Chromatographic Analysis for Classifying Medical and Industrial Cannabis.

Gas chromatography (GC) techniques for analyzing and determining the cannabinoid profile in cannabis (Cannabis sativa L.) are widely used in standard laboratories; however, these methods may mislabel the profile when used under rapid conditions. Our study aimed to highlight this problem and optimize GC column conditions and mass spectrometry (MS) parameters to accurately identify cannabinoids in both standards and forensic samples. The method was validated for linearity, selectivity, and precision. It was observed that when tetrahydrocannabinol (Δ9-THC) and cannabidiolic acid (CBD-A) were examined using rapid GC conditions, the resulting derivatives generated identical retention times. Wider chromatographic conditions were applied. The linear range for each compound ranged from 0.02 µg/mL to 37.50 µg/mL. The R2 values ranged from 0.996 to 0.999. The LOQ values ranged from 0.33 µg/mL to 5.83 µg/mL, and the LOD values ranged from 0.11 µg/mL to 1.92 µg/mL. The precision values ranged from 0.20% to 8.10% RSD. In addition, forensic samples were analyzed using liquid chromatography (HPLC-DAD) in an interlaboratory comparison test, with higher CBD and THC content than GC-MS determination (p < 0.05) in samples. Overall, this study highlights the importance of optimizing GC techniques to avoid mislabeling cannabinoids in cannabis samples.

Non-enzymatic browning due to storage is reduced by using clarified lemon juice as acidifier in industrial-scale production of canned peach halves.

Non-enzymatic browning (NEB) in canned peach halves in syrup during storage was investigated. Absorbance at 420 nm (A420), colorimetric parameters (CIELab, TCD and La/b), fructose, glucose and sucrose, total sugar, organic acids, ascorbic acid (AA), dehydroascorbic acid, and 2,3-diketogulonic acid were used to estimate the extent of NEB during 1 year of storage at 30 °C and the relationships between each of these parameters and A420 were established. The investigation was carried out to explore the possibility of replacing the E330 commonly used as acidifier by turbid or clarified lemon juice (TLJ or CLJ) to obtain a product having good nutrition with better retention of quality. The a, La/b, glucose and fructose were positively correlated with A420 and all proved to be good indicators of browning development. Overall results showed that replacement of acidifier E330 with CLJ for controlling pH in canned peach halves in syrup had some advantages.

The promiscuous and synergic molecular interaction of polyphenols in bactericidal activity: an opportunity to improve the performance of antibiotics?

Plant polyphenols are a potential source of new antimicrobial molecules against bacteria because most newly developed antimicrobial agents do not improve the clinical management of infectious diseases. The potential synergism between the major polyphenolic compounds present in a Cistus salviifolius extract, which was characterized by HPLC-ESI-MS/MS, was investigated by the isobole method and the fractional inhibitory concentration index determination. Pairwise combinations of selected flavonoids and ellagitannins present in C. salviifolius extract were assayed against the in vitro growth of Staphylococcus aureus. Some combinations revealed synergic effects, resulting in a reduction of the minimum inhibitory concentration required to inhibit 50% growth (MIC50 ) up to 20 times lower as compared with the individual compounds. Some of the combinations exhibited MIC50 values close to drug potency level (0.5-1 µg/mL). Punicalagin and myricetin were the major contributors in the combinations. The proportion between the compounds in the synergic mixtures is crucial and may explain the superior antimicrobial activity displayed by this extract when compared with other botanical extracts. The rational optimization of these combinations could lead to the design of potent antimicrobial phytopharmaceuticals, which may improve the performance of current antibiotics, taking advantage of the multi-targeted and synergic molecular interactions of selected polyphenols.

Autoinducers modulation as a potential anti-virulence target of bacteria by phenolic compounds.

OBJECTIVE: The goal of this study was to determine the effects of phenolic extracts from grape (GrPE), pomegranate (PoPE), and persimmon (PePE) by-products on bacterial virulence activities such as biofilms, motility, energy-dependent efflux pumps, and ß-lactamase activity, which are modulated primarily by quorum sensing (QS), defining their potential applications. METHOD: The microdilution method was used to determine the minimum inhibitory concentration (MIC) and sub-inhibitory concentrations (SICs) of the extracts against reference pathogenic bacteria. The antibacterial mode of action was determined by labelling bacterial cells in in vivo cell-tracking experiments. RESULTS: Antibiograms showed that PoPE inhibited bacteria at lower concentrations, and PePE had a stronger effect against Klebsiella pneumoniae. Both extracts caused significant cell membrane damage (CMD), whereas GrPE did not. At SICs, all extracts showed anti-QS activity, especially PePE, which inhibited violacein and pyocyanin production at 1/128 × MIC. Additionally, QS autoinducers found in Chromobacterium violaceum and Pseudomonas aeruginosa were modulated by the extracts; PePE showed the highest modulation. Antibiofilm assays revealed that GrPE, at MIC and 2 × MIC, acted as a potent antibiofilm agent against biofilms of Pseudomonas putida, Bacillus cereus, and Staphylococcus aureus, which was related to disruption of swarming motility by GrPE. All extracts, especially PoPE, exerted a potent effect against the activation of efflux pumps of P. aeruginosa as well as ß-lactamase activity in K. pneumoniae. CONCLUSION: Results suggest that the anti-virulence potential of the extracts may be related to their effect as extracellular autoinducer modulators. This study allowed to define potential applications of these extracts.

Green Technologies for Persimmon By-Products Revalorisation as Sustainable Sources of Dietary Fibre and Antioxidants for Functional Beverages Development.

The use of green technologies such as ultrasound and natural deep eutectic solvents (NADES) for revalorisation of food and agricultural by-products represents a sustainable way to tackle waste and promote a healthier environment while delivering much-needed functional food ingredients for an increasingly unhealthy population. The processing of persimmon (Diospyros kaki Thunb.) generates large amounts of by-products rich in fibre-bound bioactive phytochemicals. This paper assessed the extractability of bioactive compounds through NADES and the functional properties of the persimmon polysaccharide-rich by-products to evaluate their suitability to be used as functional ingredients in commercial beverages. Although higher amounts of carotenoids and polyphenols were extracted after eutectic treatment vs. conventional extraction (p < 0.05), the fibre-bound bioactives remained abundant (p < 0.001) in the resulting persimmon pulp by-product (PPBP) and persimmon pulp dietary fibre (PPDF), showing also a strong antioxidant activity (DPPH•, ABTS•+ assays) and an improved digestibility and fibre fermentability. The main structural components of PPBP and PPDF are cellulose, hemicellulose and pectin. PPDF-added dairy-based drink showed more than 50% of preference over the control among panellists and similar acceptability scores to the commercial ones. Persimmon pulp by-products represent sustainable source of dietary fibre and bioactives and are suitable candidates to develop functional ingredients for food industry applications.

Potential of Plant Exosome Vesicles from Grapefruit (Citrus × paradisi) and Tomato (Solanum lycopersicum) Juices as Functional Ingredients and Targeted Drug Delivery Vehicles.

Plant-derived extracellular vesicles (PEVs) have gained attention as promising bioactive nutraceutical molecules; their presence in common fruit juices has increased their significance because human interaction is inevitable. The goal of this study was to evaluate the potential of PEVs derived from grapefruit and tomato juices as functional ingredients, antioxidant compounds, and delivery vehicles. PEVs were isolated using differential ultracentrifugation and were found to be similar in size and morphology to mammalian exosomes. The yield of grapefruit exosome-like vesicles (GEVs) was higher than that of tomato exosome-like vesicles (TEVs), despite the latter having larger vesicle sizes. Furthermore, the antioxidant activity of GEVs and TEVs was found to be low in comparison to their juice sources, indicating a limited contribution of PEVs to the juice. GEVs showed a higher efficiency in being loaded with the heat shock protein 70 (HSP70) than TEVs, as well as a higher efficiency than TEV and PEV-free HSP70 in delivering HSP70 to glioma cells. Overall, our results revealed that GEVs present a higher potential as functional ingredients present in juice and that they exert the potential to deliver functional molecules to human cells. Although PEVs showed low antioxidant activity, their role in oxidative response in cells should be further addressed.

Antimicrobial activity of food-compatible plant extracts and chitosan against naturally occurring micro-organisms in tomato juice.

BACKGROUND: Chitosan (AC) and five hydroalcoholic extracts from Lithospermum erythrorhizon (SE), Rheum palmatum (RE), Thymus vulgaris (AT), Lippia citriodora (PLX) and a mixture of Rosmarinus officinalis, Salvia lavandulifolia and Thymus mastichina (LA) were tested for antimicrobial activity against bacteria, yeasts and filamentous fungi using two broth dilution methods. The effects of adding single extracts on naturally occurring micro-organisms and sensory qualities of raw tomato juice were also evaluated. RESULTS: SE extract exhibited the strongest activity, with minimum inhibitory concentrations (MICs) of 100-400 µg mL⁻¹ for Gram-positive and 1600-3200 µg mL⁻¹ for Gram-negative bacteria. Enterobacter aerogenes showed the greatest susceptibility to AC (MIC 1600 µg mL⁻¹). Lethal effects of extracts and AC were achieved at a minimum bactericidal concentration (MBC)/MIC ratio of 2 in 88% of assays. SE and RE extracts and AC also exhibited antifungal effect against yeasts, but they had no activity on filamentous fungi. Control and 100 mg L⁻¹ SE-added tomato juices did not differ in acceptance, but this SE concentration was not effective in the control of microbial load throughout cold storage. CONCLUSION: Results confirm the antimicrobial potential of the plant extracts, but additional research is needed until the agents responsible for the activities have been determined in order to use them as natural constituents of multiple-barrier food preservation systems.

A systematic study of the polyphenolic composition of aqueous extracts deriving from several Cistus genus species: evolutionary relationship.

INTRODUCTION: Cistaceae is a large family of shrubs widely spread over the Mediterranean area. It includes Helianthemum, Halimium and Cistus genus. Cistus genus contains approximately 20 species distributed in three subgenus. The essential oil of Cistus species has been thoroughly studied, but the polyphenolic composition of the aerial parts of the different Cistus species needs further characterisation. OBJECTIVE: To perform a comparative analysis of the qualitative and quantitative polyphenolic composition of the aerial parts of the most commonly distributed Spanish Cistus species in order to find a relationship between chemotype and subgenus. METHODOLOGY: Thirteen aqueous extracts derived from 10 different Cistus species were analysed by using HPLC with diode array-detection coupled to electrospray ion trap mass spectrometry technique (HPLC-DAD-ESI-MS/MS). Their major compounds were identified and ellagitannins were quantified. Principal component analysis (PCA) was performed on the most relevant compounds to find out the statistical association between chemotype and variety. RESULTS: Three main groups of compounds were found, i.e. ellagitannins, flavonoids and phenolic acids derivatives. The polyphenolic profile was specific for each species, although the abundance of some compounds also varied depending on the soil type. Whereas C. ladanifer, C. salviifolius, C. populifolius and C. libanotis were specially rich in ellagitannins, C. clusii, C. laurifolius and C. monspeliensis contained significant amounts of flavonoids and much less ellagitannins. In contrast, C. crispus, C. incanus and C. albidus showed a polyphenolic profile mostly based on flavonoids. PCA analysis showed a strong relationship between Cistus subgenus and its chemotype based on the most relevant water-soluble polyphenolic compounds. CONCLUSIONS: Chemical composition of the leaves' aqueous extracts from plants belonging to the Cistus genus is strongly related to their subgenus, in agreement to previous taxonomical and phylogenetic divisions. In contrast, soil and climate are less influencing factors. Leucocistus and Halimioides subgenus showed a higher content in ellagitannins. However, Cistus subgenus had higher flavonoid content.

Phytochemical characterisation for industrial use of pomegranate (Punica granatum L.) cultivars grown in Spain.

BACKGROUND: The study of 15 pomegranate cultivars was carried out in order to demonstrate the wide diversity among the quality of Spanish pomegranates, and compared to 'Wonderful' cultivars, which are used worldwide for industrial purposes. Juice yield, quality parameters, organic acids and sugars, colour, vitamin C, ellagic acid, punicalagins, anthocyanins, total phenolic content, antioxidant capacity, and sensory properties were measured. RESULTS: A considerable variation was found between 'Wonderful' and pomegranates of Spanish origin, according to both cluster analysis and principal component analysis. 'Wonderful' juices displayed large antioxidant activity and a polyphenol content with very high acidity. In contrast, 'Mollar de Elche' showed fewer anthocyanins although it had very superior organoleptical properties. In addition to a high content in ellagitannins, 'Valenciana' juices had exclusive colour parameters. CONCLUSION: These useful results can be employed by the juice processing industry to select those properties more interesting for the development of pomegranate juices that are organoleptically attractive and rich in bioactive compounds.

Potential of Persimmon Dietary Fiber Obtained from Byproducts as Antioxidant, Prebiotic and Modulating Agent of the Intestinal Epithelial Barrier Function.

Appropriate nutrition targets decrease the risk of incidence of preventable diseases in addition to providing physiological benefits. Dietary fiber, despite being available and necessary in balanced nutrition, are consumed at below daily requirements. Food byproducts high in dietary fiber and free and bonded bioactive compounds are often discarded. Herein, persimmon byproducts are presented as an interesting source of fiber and bioactive compounds. The solvent extraction effects of dietary fiber from persimmon byproducts on its techno- and physio-functional properties, and on the Caco-2 cell model after being subjected to in vitro gastrointestinal digestion and probiotic bacterial fermentation, were evaluated. The total, soluble, and insoluble dietary fiber, total phenolic, carotenoid, flavonoid contents, and antioxidant activity were determined. After in vitro digestion, low quantities of bonded phenolic compounds were detected in all fiber fractions. Moreover, total phenolic and carotenoid contents, as well as antioxidant activity, decreased depending on the extraction solvent, whereas short chain fatty acids production increased. Covalently bonded compounds in persimmon fiber mainly consisted of hydroxycinnamic acids and flavanols. After probiotic bacterial fermentation, few phenolic compounds were determined in all fiber fractions. Results suggest that persimmon's dietary fiber functional properties are dependent on the extraction process used, which may promote a strong probiotic response and modulate the epithelial barrier function.

Carotenoids from Persimmon (Diospyros kaki Thunb.) Byproducts Exert Photoprotective, Antioxidative and Microbial Anti-Adhesive Effects on HaCaT.

Persimmon (Diospyros kaki Thunb.) fruits are a remarkable source of carotenoids, which have shown protective effects against UV radiation in bacteria, fungi, algae, and plants. The aim of this study was to analyze the photoprotection provided by an acetone extract, rich in carotenoids and obtained from byproducts derived from the persimmon juice industry, against UV-induced cell death in the keratinocyte HaCaT cell line. For this purpose, the cytotoxicity and phototoxicity of carotenoid extract, as well as its intracellular reactive oxygen species (ROS) scavenging and anti-adhesive activities towards HaCaT cells, were evaluated. The in vitro permeation test provided information about the permeability of the carotenoid extract. Persimmon extracts, rich in carotenoids (PEC), were absorbed by HaCaT keratinocyte cells, which reduced the UV-induced intracellular ROS production in treated cells. Thus, PEC exerted a photoprotective and regenerative effect on UV-irradiated HaCaT cells, and this protection was UV dose-dependent. No cytotoxic effect was observed in HaCaT cultures at the concentration tested. PEC treatment also stimulated the adhesion capacity of skin microbiome to HaCaT cells, while exhibiting a significant anti-adhesive activity against all tested pathogens. In conclusion, PEC showed potential for use as a functional ingredient in skin-care products.

Physicochemical characterization of pomegranate wines fermented with three different Saccharomyces cerevisiae yeast strains.

Three commercial Saccharomyces cerevisiae yeast strains: Viniferm Revelación, Viniferm SV and Viniferm PDM were evaluated for the production of pomegranate wine from a juice coupage of the two well-known varieties Mollar and Wonderfull. Further malolactic fermentation was carried out spontaneously. The same fermentation patterns were observed for pH, titratable acidity, density, sugar consumption, and ethanol and glycerol production. Glucose was exhausted while fructose residues remained at the end of alcoholic fermentation. A high ethanol concentration (10.91 ± 0.27% v/v) in combination with 1.49 g/L glycerol was achieved. Citric acid concentration increased rapidly a 31.7%, malic acid disappeared as result of malolactic fermentation and the lactic acid levels reached values between 0.40 and 0.96 g/L. The analysis of CIEa parameter and total anthocyanin content highlights a lower degradation of monomeric anthocyanins during winemaking with Viniferm PDM yeast. The resulting wine retains a 34.5% of total anthocyanin content of pomegranate juice blend.

Physicochemical characterization of pure persimmon juice: nutritional quality and food acceptability.

Technological process for production of non-astringent persimmon (Diospyros kaki Thunb. cv. "Rojo Brillante") juice was described. The degree of fruit ripening expressed as color index (CI) varied between 12.37 and 16.33. Persimmon juice was characterized by determining physicochemical quality parameters as yield, total soluble solids (TSS), pH, titratable acidity (TA), organic acids, and main sugars. A thermal treatment of 90 ºC for 10 s was effective in controlling naturally occurring microorganisms for at least 105 d of storage without significantly affecting production of soluble brown pigments (BPs) and 5-hydroxymethyl furfural (5-HMF), total phenolic compounds (TPC), antioxidant capacity and acceptability of juice by panelists. Storage time affected all and each of the above parameters, reducing BPs, TPC and antioxidant capacity but increasing 5-HMF content. Refrigerated storage enhanced the acceptability of the juices. This information may be used by the juice industry as a starting point for production of pure persimmon juices.

Characterization of polyphenols, sugars, and other polar compounds in persimmon juices produced under different technologies and their assessment in terms of compositional variations.

Persimmon juice is emerging in the global juice market as a new wholesome commercial juice that could effectively complement a healthy diet, given the epidemiological evidence linking a diet rich in fruits and vegetables with reduced incidences of chronic diseases. However, little data are available on the persimmon-juice composition or on the effect of the technological treatment employed for its production. The present work performs a complete qualitative analytical characterization through high-performance liquid chromatography coupled to electrospray time-of-flight mass spectrometry (HPLC-DAD-ESI-TOF/MS) of the diverse persimmon juices produced under different technologies in a pilot plant (clarification, astringency removal, flash vacuum expansion, centrifugation and pasteurization) in order to evaluate the effect of the different production procedures on the polar chemical profile of persimmon juice. Persimmon-juice extracts have been found to be a source of sugars, protein derivatives, organic acids, vitamins, and polyphenols, including simple polyphenols (phenolic acids and flavonoids) and polymerized flavan-3-ols. A marked influence of processing on the composition of the juices has been noticed. Extracts 3 and 7 (undergoing the combinations of clarification and centrifugation, and astringency removal, centrifugation and pasteurization, respectively) contained more polyphenols, which may help reduce risk of chronic diseases.

Determination of piceid and resveratrol in Spanish wines deriving from Monastrell (Vitis vinifera L.) grape variety.

The presence of stilbenes in wine is becoming an important issue due to their claimed relation to a low incidence in coronary diseases and their increasing implication as cancer chemopreventive and neuroprotective agents. Total resveratrol content, quantified as glucoside and aglycone forms of resveratrol, has been determined in a survey of 45 Monastrell monovarietal Spanish red wine types (around 135 wine samples), belonging to Alicante and Bullas appellations. The average between ratio glucoside/aglycone forms of resveratrol in these wines was considerably high, ranging from 82 to 91% of resveratrol in its glycosidic form. This characteristic was observed in a high percentage of the studied wines, which were made under different winemaking procedures, and from different vintages (1995-2002). In addition, wines made using macerative fermentations with double amount of solid parts ("doble pasta") reached the highest levels of total stilbene content expressed as resveratrol equivalent, i.e., 30 mg/L (average of 18.8 mg/L). It can be concluded that high resveratrol glucoside concentration and low free isomer content can be considered characteristics of the Monastrell variety, as it happens to red wines deriving from other varieties grown at warm climates. This fact, also observed for other French and Portuguese red varieties, might play an important role in food habits involving these types of wines.

Chemical guide parameters for Punica granatum cv. 'Mollar' fruit juices processed at industrial scale.

To contribute for setting reference guideline for commercial juice from the pomegranate variety 'Mollar', chemical composition of eighteen samples directly obtained and commercialised in 2012 from three different fruit juice factories was investigated. According to the findings, the relative density of direct pomegranate juices varied between 1.061 and 1.064, which correspond to 15.15 and 15.71°Brix; titratable acidity changed between 2.6 and 2.8g/L, citric acid between 2.3 and 2.8 g/L, l-malic acid in a range of 1.3-1.4 g/L, and d-isocitric acid at levels less than 20mg/L. Glucose values ranged from 61.4 to 65.0 g/L, whereas fructose displayed values between 65.3 and 68.0 g/L. The predominant mineral was potassium (2,400-2,900 mg/L), followed by phosphorous, magnesium, calcium and sodium at levels of 81-89 mg/L, 17.6-28.5mg/L, 5.8-7.5mg/L and 4.3-5.3mg/L, respectively. Chemical determinations of anthocyanin and ellagitannin profiles and amino acids contents were also carry out. Concentrations of anthocyanins in commercialised samples were Cy3,5dG (19.30 ± 3.47 mg/L), followed by Dp3,5dG (17.87 ± 6.74 mg/L) and Cy3G (12.91 ± 6.32 mg/L). Punicalagin levels ranged between 503.70 and 762.85 mg/L, punicalins between 239.9 and 364.5mg/L, and free ellagic acid level was typically between 268.67 and 389.64 mg/L. The juice samples exhibited high amount of total phenolics (1,136-3,581 mg/L) as well as high ABTS radical scavenging activity (18-31 mmol Trolox/L).

Chemical guide parameters for Spanish lemon (Citrus limon (L.) Burm.) juices.

To contribute for setting reference guideline for commercial juice extracted from the Spanish lemon varieties, chemical composition of 92 direct and 92 reconstituted samples were investigated. In direct lemon juice, titratable acidity was 52.4 g/L, being the citric acid the main component. Glucose, fructose and sucrose concentrations were 7.9, 7.3 and 4.5 g/L, respectively. Predominant mineral was potassium (1264.2mg/L), followed by phosphorous (306 mg/L), calcium (112 mg/L) and magnesium (92.6 mg/L). Hesperidin ranged from 257 to 484.8 mg/L, while water soluble pectins varied between 164.8 and 550 mg/L. Similar values were obtained in reconstituted lemon juice. There are different parameters that did not reach or exceeded the limits proposed by the European Association of the Industry of Juices and Nectars. These levels should be taken into account to modify the present reference guideline and that Spanish lemon juices are not discarded for to have lower or bigger values.

Approaches to understanding the contribution of anthocyanins to the antioxidant capacity of pasteurized pomegranate juices.

Influence of processing and storage conditions on anthocyanin stability and antioxidant activity of clarified and cloudy juices from arils of the 'Mollar' pomegranate variety was studied. Clarification process reduced the content of total monomeric and individual anthocyanins, and increased the antioxidant activity of pomegranate juice. Thermal treatments (65 and 90 °C for 30 or 5 s) decreased the percentage of polymeric anthocyanin form, increasing on the contrary the monomeric one. In any case, storage temperature was the main factor affecting all the parameters tested. Cyanidin 3-O-glucoside (Cy3G) was more instable than delphinidin 3,5-di (Dp3,5dG) and cyaniding 3,5-diglucosides (Cy3,5dG). A linear relationship was observed between oxygen radical absorbance capacity (ORAC) and total monomeric anthocyanins, suggesting that they contributed strongly to the antioxidant capacity. Results presented in this study show that hurdle technology (heating plus refrigeration) may help to reduce anthocyanin degradation in pasteurized pomegranate juice, avoiding a dramatic impact on its colour and preserving the beneficial effects of this specific bioactive compounds on human health.

Changes on indigenous microbiota, colour, bioactive compounds and antioxidant activity of pasteurised pomegranate juice.

Juices prepared from arils of 'Mollar' pomegranates were analysed for naturally occurring microorganisms, CIE Lab colour parameters, total phenols, anthocyanins and punicalagins, ellagic acid content and antioxidant capacity before and after low-, mild- and high-temperature pasteurisations (LTPs, MTPs and HTPs): 65, 80 and 90 °C for 30 or 60s. Mean aerobic plate count (APC), yeast and mold count (YMC), and lactic acid bacteria (LAB) for fresh juices were 5.7, 5.36 and 4.0 log CFU/mL, respectively. MTPs and HTPs were sufficiently effective to decrease APCs to nil or negligible levels. An increase in CIE a values and decrease in CIE b values were the characteristic colour changes in heat-treated juices. The effect of pasteurisations showed that total phenols, punicalagins and ellagic acid were not much affected by thermal processing. Total anthocyanin content and antioxidant capacity were substantially and significantly influenced by the heat treatment applied. A linear relationship was observed between Trolox equivalent antioxidant capacity (TEAC) values and total anthocyanins, suggesting that they contributed strongly to the antioxidant capacity of pomegranate juice.