Exploring the potential of phenolic and antioxidant compounds in new Rosaceae fruits.

BACKGROUND: Rosaceae fruits have been used in traditional medicine for the prevention and treatment of diseases. However, these fruits have not extensively been studied regarding their phenolic composition. Thus, this research focuses on the determination of phenolic compounds by high-performance liquid chromatography electrospray ionization time-of-flight mass spectrometry, flavan-3-ols by high-performance liquid chromatography with fluorescence detection, and the antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power of the fruits of five species of genera Crataegus and Sorbus (Rosaceae). RESULTS: We found a total of 71 phenolic compounds from which 30 were identified in these berries for the first time. Crataegus monogyna and Crataegus laciniata revealed higher total phenolic and flavan-3-ol contents than the other species and the highest antioxidant activities. CONCLUSIONS: Therefore, the fruits evaluated have demonstrated to be important sources of bioactive compounds with huge potential for being used in nutraceutical or food scopes. Additional studies could be needed to evaluate the influence of the different production areas on the phenolic content. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Setup of an Ultrasonic-Assisted Extraction to Obtain High Phenolic Recovery in Crataegus monogyna Leaves.

Hawthorn leaves are a rich source of phenolic compounds that possess beneficial activities for human health. Ultrasonic-assisted extraction (UAE) is an extraction technique frequently used for the isolation of phenolic compounds in plants. Thus, in this study, a Box-Behnken design was used to optimize UAE conditions such as the percentage of acetone, the extraction time and solvent-to-solid ratio (v/w) in order to obtain the maximum content of total compounds by Folin-Ciocalteu and the maximum in vitro antioxidant activity by DPPH, ABTS and FRAP assays in Crataegus monogyna leaves. The optimum conditions to obtain the highest total phenolic content and antioxidant activities were 50% acetone, 55 min and 1/1000 (w/v). A total of 30 phenolic compounds were identified and quantified in C. monogyna leaf extract obtained at these optimum UAE conditions. HPLC-MS allows the identification and quantification of 19 phenolic compounds and NP-HPLC-FLD analyses showed the presence of 11 proanthocyanidins. According to the results, the most concentrated phenolic compounds in C. monogyna leaf extract obtained at optimum UAE conditions were phenolic acid derivatives such as protocatechuic acid-glucoside, dihydroxy benzoic acid pentoside and chlorogenic acid, flavones such as 2″-O-rhamnosyl-C-hexosyl-apigenin, flavonols such as hyperoside and isoquercetin and proanthocyanidins such as monomer and dimer. As a result, the optimized UAE conditions could be used to obtain an extract of C. monogyna leaves enriched with phenolic compounds.

Zygophyllum album saponins prevent atherogenic effect induced by deltamethrin via attenuating arterial accumulation of native and oxidized LDL in rats.

The current study aimed to examine, for the first time, the relationship between exposure to deltamethrin (DLM) and atherogenic lipid profile disorders in adult Wistar rats, as well as, to verify the mechanism of the beneficial role of Zygophyllum album leaves extracts (ZALE). The experimental study was assessed using DLM (4 mg/kg b.w) either alone or co administered with ZALE (400 mg/kg b.w) orally for 90 days in rats. RP-HPLC-DAD-ESI-QTOF-MS was used to identify the bioactive metabolites present in ZALE. Plasmatic and aortic total cholesterol (TC), LDL-cholesterol (LDL-C), native LDL (LDL-apo B-100) and oxidized LDL (ox-LDL) were evaluated using auto-analyzer and a sandwich ELISA, respectively. The protein expressions of LDLR (native LDL receptor) and CD36 (Scavenger receptor class B) were evaluated in aorta or liver with a Western blot. The pathology has been confirmed with lipid stain (Oil Red O). Phytochemicals analysis revealed the presence of fifteen saponins in ZALE. Rats intoxicated with DLM revealed a significant increase in plasmatic and aortic lipid profile (TC, LDL-C, LDL-apo B-100 and ox-LDL), as well as, the concentration of the plasmatic cytokines include TNF-α, IL-2 and IL-6, compared to control. Hepatic native LDL and aortic CD36 receptor expression were increased in DLM treated group, however aortic LDL-R does not present any modification, when compared to control. The detected disturbances in lipid parameters were supported by Oil Red O applied. Due to their antioxidant activity, the bioactive compounds in ZALE as powerful agents able to prevent the pro-atherogenic effect observed in DLM-treated animals. These metabolites modulated most of inflammatory markers, prevented accumulation of lipid and lipoprotein biomarkers, regulated the major receptor regulators of hepatic cholesterol metabolism, as well as normalize lipid distribution in liver and aorta tissue.

Zygophyllum album leaves extract prevented hepatic fibrosis in rats, by reducing liver injury and suppressing oxidative stress, inflammation, apoptosis and the TGF-ß1/Smads signaling pathways. Exploring of bioactive compounds using HPLC-DAD-ESI-QTOF-MS/MS.

Zygophyllum album is traditionally used against many illnesses, such as liver disease. The present study investigated the bioactive compounds in methanol extract of Z. album (MEZA) using HPLC-DAD-ESI-QTOF-MS/MS and explored its possible antioxidative, anti-inflammatory, anti-apoptotic, and hepatoprotective effect. Twelve phenolic compounds were identified; isorhamnetin-3-O-rutinoside being the main one was the main composite (144.6 mg/100 g dm). Results showed that MEZA reduced significantly the biochemical markers (AST, ALT, LDH and ALP), and the hepatic oxidative stress indicators (MDA, PC, SOD, CAT, and GPx) in deltamethrin (DLM)-treated rats. Moreover, MEZA limited the inflammatory responses through downregulation of NF-κB gene, which suppressed the production of proinflammatory cytokines (TNF-α, IL-1ß, IL-6). Furthermore, Z. album reduced DLM-induced apoptosis by attenuating caspase 3 and p53 mRNA activation. MEZA treatment also alleviated upregulation of α-SMA, type I collagen, and TGF-ß1 mRNA in the liver. The possible antifibrotic effect of MEZA was clearly demonstrated by the histopathology examination, using Masson's Trichrome and Sirius Red stainings. Therefore, the current study suggested that the bioactive compounds of Z. album possessed antifibrotic effect against DLM-induced hepatic fibrosis, by protecting liver tissue, and inhibiting oxidative stress, inflammation, apoptosis and the TGF-ß1/Smads signaling pathways.

Leaf removal at veraison stage differentially affects qualitative attributes and bioactive composition of fresh and dehydrated grapes of two indigenous Cypriot cultivars.

BACKGROUND: The effect of leaf removal on postharvest performance of dehydrated grapes has been poorly analyzed. The aim of this study was to evaluate the effect of leaf removal at veraison stage on the metabolites of fresh and dehydrated grapes of two indigenous Cypriot cultivars ('Mavro' and 'Xynisteri'), which are destined for the production of 'Commandaria', a protected designation of origin (PDO) premium sweet wine. RESULTS: Leaf removal led to a reduction of soluble solids, titratable acidity, aroma potential and most of the phenolic groups in the musts of both cultivars. Dehydration led to a significant increase in all of these parameters in both cultivars, being more pronounced in cv. 'Mavro'. Interestingly, leaf removal indicated differential response in the dehydrated product of the cultivars examined; liquid chromatography (LC-DAD-qTOF-MS) data showed a significant decrease in phenolic compounds in 'Xynisteri' must (from 66.73 to 44.15 mg L-1 ), while 'Mavro' must registered similar values of phenolic compounds (from 94.78 to 96.72 mg L-1 ), but with a different distribution among phenolic groups. Intriguingly, flavonols and flavan-3-ols that present significant health-promoting properties showed higher concentrations in the must from dehydrated 'Mavro' grapes that were subjected to preharvest leaf removal. CONCLUSIONS: Leaf removal at veraison stage followed by sun drying differentially affected the chemical composition of the examined cultivars. This preharvest application followed by postharvest dehydration was beneficial for cv. 'Mavro', while this was not the case for cv. 'Xynisteri'. The effects of leaf removal in other developmental stages (i.e. pre/post bloom stage) need to be dissected with the ultimate goal of providing an end product with high bioactive content. © 2018 Society of Chemical Industry.

Establishment of pressurized-liquid extraction by response surface methodology approach coupled to HPLC-DAD-TOF-MS for the determination of phenolic compounds of myrtle leaves.

Myrtus communis L. (myrtle) is native to the Mediterranean region and Western Asia. Its leaves have demonstrated its potential effect towards different bioactivities like anti-diabetic, anti-diarrheic, anti-ulcer, anti-cancer, among others. These activities have been associated with its phenolic content. In this sense, the aim of this work has been to develop a new pressurized-liquid extraction procedure (PLE), by using a response surface methodology (RSM), to evaluate the phenolic composition from myrtle leaves by HPLC-DAD-TOF-MS. Previously, different solvents such as methanol, ethanol, and acetone/water mixtures were tested by using ultrasound-assisted extraction (UAE) in order to select the most suitable one. Subsequently, a Box-Behnken design (BBD) was performed according to the effect of ethanol/water ratio (50, 75, and 100% (v/v)), temperature (50, 125, and 200 °C), and extraction time (5, 18, and 30 min). The optimal conditions achieved with the established method were 71% ethanol/water, 137 °C, and 19 min. The analysis of the obtained extracts by HPLC-DAD-TOF-MS allowed the characterization of 15 new compounds in myrtle leaves. Finally, high amounts of gallic and ellagic acid were found in the optimized PLE extracts (3.31 ± 0.03 and 3.88 ± 0.09 mg/g leaf dry weight (d.w.), respectively), and PLE reported greater recovery of total phenolic compounds than UAE (30 ± 1 and 22.4 ± 0.6 mg/g leaf d.w., respectively).

Characterization of bioactive compounds of Annona cherimola L. leaves using a combined approach based on HPLC-ESI-TOF-MS and NMR.

Annona cherimola Mill. (cherimoya) has widely been used as food crop. The leaves of this tree possess several health benefits, which are, in general, attributed mainly to its bioactive composition. However, literature concerning a comprehensive characterization based on a combined approach, which consists of nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS), from these leaves is scarce. Thus, the aim of this work was to study the polar profile of full extracts of cherimoya leaves by using these tools. Thus, a total of 77 compounds have been characterized, 12 of which were identified by both techniques. Briefly, 23 compounds were classified as amino acids, organic acids, carbohydrates, cholines, phenolic acid derivatives, and flavonoids by NMR, while 66 metabolites were divided into sugars, amino acids, phenolic acids and derivatives, flavonoids, phenylpropanoids, and other polar compounds by HPLC-TOF-MS. It is worth mentioning that different solvent mixtures were tested and the total phenolic content in the extracts quantified (TPC via HPLC-TOF-MS). The tendency observed was EtOH/water 80/20 (v/v) (17.0 ± 0.2 mg TPC/g leaf dry weight (d.w.)) ≥ acetone/water 70/30 (v/v) (16.1 ± 0.7 mg TPC/g leaf d.w.) > EtOH/water 70/30 (v/v) (14.0 ± 0.3 mg TPC/g leaf d.w.) > acetone/water 80/20 (v/v) (13.5 ± 0.4 mg TPC/g leaf d.w.). Importantly, flavonoids derivatives were between 63 and 76% of the TPC in those extracts. Major compounds were sucrose, glucose (α and ß), and proline, and chlorogenic acid and rutin for NMR and HPLC-TOF-MS, respectively. Graphical abstract The combined use of LC-HRMS and NMR is a potential synergic combination for a comprehensive metabolite composition of cherimoya leaves.

Recent Advances in Phospholipids from Colostrum, Milk and Dairy By-Products.

Milk is one of the most important foods for mammals, because it is the first form of feed providing energy, nutrients and immunological factors. In the last few years, milk lipids have attracted the attention of researchers due to the presence of several bioactive components in the lipid fraction. The lipid fraction of milk and dairy products contains several components of nutritional significance, such as ω-3 and ω-6 polyunsaturated fatty acids, CLA, short chain fatty acids, gangliosides and phospholipids. Prospective cohort evidence has shown that phospholipids play an important role in the human diet and reinforce the possible relationship between their consumption and prevention of several chronic diseases. Because of these potential benefits of phospholipids in the human diet, this review is focused on the recent advances in phospholipids from colostrum, milk and dairy by-products. Phospholipid composition, its main determination methods and the health activities of these compounds will be addressed.

Comparison of Two Stationary Phases for the Determination of Phytosterols and Tocopherols in Mango and Its By-Products by GC-QTOF-MS.

Two different gas chromatography coupled to quadrupole-time of flight mass spectrometry (GC-QTOF-MS) methodologies were carried out for the analysis of phytosterols and tocopherols in the flesh of three mango cultivars and their by-products (pulp, peel, and seed). To that end, a non-polar column ((5%-phenyl)-methylpolysiloxane (HP-5ms)) and a mid-polar column (crossbond trifluoropropylmethyl polysiloxane (RTX-200MS)) were used. The analysis time for RTX-200MS was much lower than the one obtained with HP-5ms. Furthermore, the optimized method for the RTX-200MS column had a higher sensibility and precision of peak area than the HP-5ms methodology. However, RTX-200MS produced an overlapping between ß-sitosterol and Δ5-avenasterol. Four phytosterols and two tocopherols were identified in mango samples. As far as we are concerned, this is the first time that phytosterols have been studied in mango peel and that Δ5-avenasterol has been reported in mango pulp. α- and γ-tocopherol were determined in peel, and α-tocopherol was the major tocopherol in this fraction (up to 81.2%); however, only α-tocopherol was determined in the pulp and seed. The peel was the fraction with the highest total concentration of phytosterols followed by seed and pulp, and "Sensación" was the cultivar with the highest concentration of total phytosterols in most cases. There were no significant differences between quantification of tocopherols with both columns. However, in most cases, quantification of phytosterols was higher with RTX-200MS than with HP-5ms column.

Health Effects of Psidium guajava L. Leaves: An Overview of the Last Decade.

Today, there is increasing interest in discovering new bioactive compounds derived from ethnomedicine. Preparations of guava (Psidium guajava L.) leaves have traditionally been used to manage several diseases. The pharmacological research in vitro as well as in vivo has been widely used to demonstrate the potential of the extracts from the leaves for the co-treatment of different ailments with high prevalence worldwide, upholding the traditional medicine in cases such as diabetes mellitus, cardiovascular diseases, cancer, and parasitic infections. Moreover, the biological activity has been attributed to the bioactive composition of the leaves, to some specific phytochemical subclasses, or even to individual compounds. Phenolic compounds in guava leaves have been credited with regulating blood-glucose levels. Thus, the aim of the present review was to compile results from in vitro and in vivo studies carried out with guava leaves over the last decade, relating the effects to their clinical applications in order to focus further research for finding individual bioactive compounds. Some food applications (guava tea and supplementary feed for aquaculture) and some clinical, in vitro, and in vivo outcomes are also included.

HPLC-DAD-q-TOF-MS as a powerful platform for the determination of phenolic and other polar compounds in the edible part of mango and its by-products (peel, seed, and seed husk).

Free and bound phenolic and other polar compounds in mango edible fraction and its by-products (peel, seed, and seed husk) have been determined by HPLC-DAD-ESI-qTOF-MS. This analytical technique has demonstrated to be a valuable platform for the identification and quantification of these compounds in mango. In fact, UV-Vis and mass spectra data allowed the determination of 91 free compounds and 13 bound (cell wall linked) compounds taking into account the four fractions of mango. To our knowledge, this is the first time that mango seed husk has been studied regarding its phenolic compounds. The method proposed showed LODs between 0.006 and 0.85 µg/mL and accuracy ranged from 94.8 and 100.7%. Mango peel presented the highest concentration of free polar compounds followed by seed, pulp, and seed husk. It is also important to highlight that bound phenolic compounds had never been determined in mango pulp, seed, and seed husk before. Furthermore, ellagic acid was the most abundant bound compound in the four mango fractions analyzed. These results show that mango pulp and its by-products are a good source of phenolic and other polar compounds. In particular, mango seed contains a high total concentration of ellagic acid (650 mg/100 g dry weight).

Exploratory Characterization of Phenolic Compounds with Demonstrated Anti-Diabetic Activity in Guava Leaves at Different Oxidation States.

Psidium guajava L. is widely used like food and in folk medicine all around the world. Many studies have demonstrated that guava leaves have anti-hyperglycemic and anti-hyperlipidemic activities, among others, and that these activities belong mainly to phenolic compounds, although it is known that phenolic composition in guava tree varies throughout seasonal changes. Andalusia is one of the regions in Europe where guava is grown, thus, the aim of this work was to study the phenolic compounds present in Andalusian guava leaves at different oxidation states (low, medium, and high). The phenolic compounds in guava leaves were determined by HPLC-DAD-ESI-QTOF-MS. The results obtained by chromatographic analysis reported that guava leaves with low degree of oxidation had a higher content of flavonols, gallic, and ellagic derivatives compared to the other two guava leaf samples. Contrary, high oxidation state guava leaves reported the highest content of cyanidin-glucoside that was 2.6 and 15 times higher than guava leaves with medium and low oxidation state, respectively. The QTOF platform permitted the determination of several phenolic compounds with anti-diabetic properties and provided new information about guava leaf phenolic composition that could be useful for nutraceutical production.

From Olive Fruits to Olive Oil: Phenolic Compound Transfer in Six Different Olive Cultivars Grown under the Same Agronomical Conditions.

Phenolic compounds are responsible of the nutritional and sensory quality of extra-virgin olive oil (EVOO). The composition of phenolic compounds in EVOO is related to the initial content of phenolic compounds in the olive-fruit tissues and the activity of enzymes acting on these compounds during the industrial process to produce the oil. In this work, the phenolic composition was studied in six major cultivars grown in the same orchard under the same agronomical and environmental conditions in an effort to test the effects of cultivars on phenolic composition in fruits and oils as well as on transfer between matrices. The phenolic fractions were identified and quantified using high-performance liquid chromatography-diode array detector-time-of-flight-mass spectrometry. A total of 33 phenolic compounds were determined in the fruit samples and a total of 20 compounds in their corresponding oils. Qualitative and quantitative differences in phenolic composition were found among cultivars in both matrices, as well as regarding the transfer rate of phenolic compounds from fruits to oil. The results also varied according to the different phenolic groups evaluated, with secoiridoids registering the highest transfer rates from fruits to oils. Moreover, wide-ranging differences have been noticed between cultivars for the transfer rates of secoiridoids (4.36%-65.63% of total transfer rate) and for flavonoids (0.18%-0.67% of total transfer rate). 'Picual' was the cultivar that transferred secoiridoids to oil at the highest rate, whereas 'Changlot Real' was the cultivar that transferred flavonoids at the highest rates instead. Principal-component analysis confirmed a strong genetic effect on the basis of the phenolic profile both in the olive fruits and in the oils.

Fermentation of Orange Peels by Lactic Acid Bacteria: Impact on Phenolic Composition and Antioxidant Activity.

Orange processing generates peel by-products rich in phenolic compounds, particularly flavanones like hesperidin and narirutin, offering potential health benefits. Utilizing these by-products is of significant interest in supporting Spain's circular bioeconomy. Therefore, the aim of this study was to investigate the fermentation of orange peels by different lactic acid bacteria (LAB) strains and its impact on phenolic composition and antioxidant activity. Three different LAB strains, two Lactiplantibacillus plantarum, and one Levilactobacillus brevis were utilized. The phenolic compounds were measured by HPLC-ESI-TOF-MS, and antioxidant activity was assessed using DPPH and ABTS methods. The growth of the LAB strains varied, showing initial increases followed by gradual declines, with strain-specific patterns observed. Medium acidification occurred during fermentation. A phenolic analysis revealed an 11% increase in phenolic acids in peels fermented by La. plantarum CECT 9567-C4 after 24 h, attributed to glycosylation by LAB enzymes. The flavonoid content exhibited diverse trends, with Le. brevis showing an 8% increase. The antioxidant assays demonstrated strain- and time-dependent variations. Positive correlations were found between antioxidant activity and total phenolic compounds. The results underscore the importance of bacterial selection and fermentation time for tailored phenolic composition and antioxidant activity in orange peel extracts. LAB fermentation, particularly with La. plantarum CECT 9567 and Le. brevis, holds promise for enhancing the recovery of phenolic compounds and augmenting antioxidant activity in orange peels, suggesting potential applications in food and beverage processing.

Mathematical Modelling of Convective Drying of Orange By-Product and Its Influence on Phenolic Compounds and Ascorbic Acid Content, and Its Antioxidant Activity.

Orange peel is one of the main by-products from juice processing, and is considered as a promising source of phenolic compounds with anti-carcinogenic, anti-inflammatory, anti-microbial and antioxidant properties. The drying is an essential step to ensure the storage of this by-product at an industrial level, in order to use it as a functional ingredient or as a nutraceutical. Thus, this research focuses on the evaluation of the effect of the convective air-drying process in orange by-products at three different temperatures (40, 60 and 80 °C) and air flows (0, 0.8 and 1.6 m/s) on the phenolic content (measured by HPLC-MS), the antioxidant activity (measured by DPPH, ABTS and FRAP), and the vitamin C content (measured by HPLC-UV/VIS). Moreover, the mathematical modelling of its drying kinetics was carried out to examine the orange by-product behavior. Among the tested mathematical models, the Page model reported the highest fit and the best drying conditions, which showed the lowest reductions were at 60 °C with an air flow of 1.6 m/s and taking 315 min.

Response Surface Methodology for the Optimization of Flavan-3-ols Extraction from Avocado By-Products via Sonotrode Ultrasound-Assisted Extraction.

Avocado peel and seed are the main by-products of avocado processing and are considered as promising sources of phenolic compounds with biological activities. Thus, this research focuses on the establishment, for the first time, of ultrasound-assisted extraction of flavan-3-ols with high antioxidant activity from avocado peel and seed using a sonotrode. Indeed, 2 Box-Behnken designs were performed for 15 experiments, with each design having three independent factors (ratio ethanol/water (v/v), time (min) and amplitude (%)). In both models, the responses included total procyanidins (flavan-3-ols) measured via HPLC-FLD and antioxidant activity measured via DPPH, ABTS and FRAP. The results showed that applying the sonotrode extraction method could increase flavan-3-ols recovery by 54% and antioxidant activity by 62-76% compared to ultrasound bath technology. Therefore, this technology was demonstrated to be a non-thermal, low time-consuming and scalable method that allowed the recovery of flavan-3-ols from avocado by-products that could be used as functional ingredients.

Establishment of a Sonotrode Extraction Method and Evaluation of the Antioxidant, Antimicrobial and Anticancer Potential of an Optimized Vaccinium myrtillus L. Leaves Extract as Functional Ingredient.

Vaccinium myrtillus L. (bilberry) leaves are an important by-product of berry production that may be used as a source of phenolic compounds which have a positive effect on human health. Therefore, an ultrasound-assisted extraction via sonotrode has been used for the first time to recover bioactive compounds from bilberry leaves. The extraction has been optimized using a Box-Behnken design. The influence of ethanol:water ratio (v/v), time of extraction (min) and amplitude (%) were evaluated considering total phenolic content (TPC) and antioxidant capacity (DPPH and FRAP assays) as dependent variables in a response surface methodology (RSM). Optimum values for the independent factors were 30:70 ethanol/water (v/v), 5 min of extraction and 55% amplitude. The empirical values of the independent variables using the optimized conditions were 217.03 ± 4.92 mg GAE/g d.w. (TPC), 271.13 ± 5.84 mg TE/g d.w. (DPPH) and 312.21 ± 9.30 mg TE/g d.w. (FRAP). The validity of the experimental design was confirmed using ANOVA and the optimal extract was characterized using HPLC-MS. A total of 53 compounds were tentatively identified, of which 22 were found in bilberry leaves for the first time. Among them, chlorogenic acid was the most abundant molecule, representing 53% of the total phenolic compounds identified. Additionally, the antimicrobial and anticancer activities of the optimum extract were tested. Gram-positive bacteria demonstrated high sensitivity to bilberry leaves extract in vitro, with MBC values of 6.25 mg/mL for Listeria monocytogenes, Listeria innocua and Enterococcus faecalis, and 0.8 mg/mL for Staphylococcus aureus and Bacillus cereus. Furthermore, bilberry leaves extract exerted in vitro antiproliferative activity against HT-29, T-84 and SW-837 colon tumor cells with IC50 values of 213.2 ± 2.5, 1140.3 ± 5.2 and 936.5 ± 4.6 µg/mL, respectively. Thus, this rapid ultrasound-assisted extraction method has demonstrated to be an efficient technique to obtain bilberry leaves extract with in vitro antioxidant, antimicrobial and anticancer capacities that may be useful for the food industry as natural preservative or even for the production of functional foods or nutraceuticals.

Phytochemicals Determination, and Antioxidant, Antimicrobial, Anti-Inflammatory and Anticancer Activities of Blackberry Fruits.

A comprehensive characterization of the phytochemicals present in a blackberry fruit extract by HPLC-TOF-MS has been carried out. The main compounds in the extract were ursane-type terpenoids which, along with phenolic compounds, may be responsible for the bioactivity of the extract. In vitro antioxidant capacity was assessed through Folin-Ciocalteu (31.05 ± 4.9 mg GAE/g d.w.), FRAP (637.8 ± 3.2 µmol Fe2+/g d.w.), DPPH (IC50 97.1 ± 2.4 µg d.w./mL) and TEAC (576.6 ± 8.3 µmol TE/g d.w.) assays. Furthermore, the extract exerted remarkable effects on in vitro cellular antioxidant activity in HUVEC cells at a concentration of 5 mg/mL. Antimicrobial activity of the extract was also tested. Most sensible microorganisms were Gram-positive bacteria, such as E. faecalis, B. cereus and Gram-negative E. coli (MBC of 12.5 mg/mL). IC50 values against colon tumoral cells HT-29 (4.9 ± 0.2 mg/mL), T-84 (5.9 ± 0.3 mg/mL) and SW-837 (5.9 ± 0.2 mg/mL) were also obtained. Furthermore, blackberry extract demonstrated anti-inflammatory activity inhibiting the secretion of pro-inflammatory IL-8 cytokines in two cellular models (HT-29 and T-84) in a concentration-dependent manner. These results support that blackberry fruits are an interesting source of bioactive compounds that may be useful in the prevention and treatment of different diseases, mainly related to oxidative stress.

Characterization of the Metabolic Profile of Olive Tissues (Roots, Stems and Leaves): Relationship with Cultivars' Resistance/Susceptibility to the Soil Fungus Verticillium dahliae.

Verticillium wilt of olive (VWO) is one of the most widespread and devastating olive diseases in the world. Harnessing host resistance to the causative agent is considered one of the most important measures within an integrated control strategy of the disease. Aiming to understand the mechanisms underlying olive resistance to VWO, the metabolic profiles of olive leaves, stems and roots from 10 different cultivars with varying levels of susceptibility to this disease were investigated by liquid chromatography coupled to mass spectrometry (LC-MS). The distribution of 56 metabolites among the three olive tissues was quantitatively assessed and the possible relationship between the tissues' metabolic profiles and resistance to VWO was evaluated by applying unsupervised and supervised multivariate analysis. Principal component analysis (PCA) was used to explore the data, and separate clustering of highly resistant and extremely susceptible cultivars was observed. Moreover, partial least squares discriminant analysis (PLS-DA) models were built to differentiate samples of highly resistant, intermediate susceptible/resistant, and extremely susceptible cultivars. Root models showed the lowest classification capability, but metabolites from leaf and stem were able to satisfactorily discriminate samples according to the level of susceptibility. Some typical compositional patterns of highly resistant and extremely susceptible cultivars were described, and some potential resistance/susceptibility metabolic markers were pointed out.

Development of an Effective Sonotrode Based Extraction Technique for the Recovery of Phenolic Compounds with Antioxidant Activities in Cherimoya Leaves.

The leaves of Annona cherimola Mill (cherimoya) are a potential source of phenolic compounds that have been shown to have beneficial properties. Therefore, this study focuses on establishing an ultrasonic-assisted extraction of phenolic compounds in cherimoya leaves using a sonotrode. For that purpose, a Box-Behnken design based on a response surface methodology (RSM) was used to optimize factors, such as amplitude, extraction time and solvent composition to obtain the maximum content of phenolic compounds by HPLC-MS and the maximum in-vitro antioxidant activity by DPPH, ABTS and FRAP assays in 'Fino de Jete' cherimoya leaves. The optimal conditions were 70% amplitude, 10 min and 40:60 ethanol/water (EtOH/H2O) (v/v). The results obtained under these optimum conditions by using a sonotrode were compared with those from an ultrasonic bath; briefly, recovery of phenolic compounds by sonotrode was 2.3 times higher than a bath. Therefore, these optimal conditions were applied to different varieties 'Campas', 'Fino de Jete' and 'Negrito Joven' harvested in the Tropical Coast of Granada (Spain). A total of 39 phenolic compounds were determined in these cherimoya leaf extracts, 24 phenolic compounds by HPLC-MS and 15 proanthocianidins by HPLC-FLD. 5-p-coumaroylquinic acid, lathyroside-7-O-α-l-rhamnopyranoside and quercetin hexose acetate were first identified in cherimoya leaves. The most concentrated phenolic compounds were the flavonoids, such as rutin and quercetin hexoside and proanthocyanidins including monomers. Almost no significant differences in the phenolic content in these cultivars were found (11-13 mg/g d.w. for phenolic compounds and 11-20 mg/g d.w. for proanthocyanidins). In addition, sonotrode ultrasonic-assisted extraction has been shown to be an efficient extraction technique in the phenolic recovery from cherimoya leaves that could be implemented on an industrial scale.