One-Step Extraction of Olive Phenols from Aqueous Solution Using ß-Cyclodextrin in the Solid State, a Simple Eco-Friendly Method Providing Photochemical Stability to the Extracts.

The extraction of phenolic compounds from olive mill wastes is important, not only to avoid environmental damages, but also because of the intrinsic value of those biophenols, well-known for their high antioxidant potential and health benefits. This study focuses on tyrosol (Tyr) and hydroxytyrosol (HT), two of the main phenolic compounds found in olive mill wastes. A new, simple, and eco-friendly extraction process for the removal of phenolic compounds from aqueous solutions using native ß-cyclodextrin (ß-CD) in the solid state has been developed. Several ß-CD/biophenol molar ratios and biophenol concentrations were investigated, in order to maintain ß-CD mostly in the solid state while optimizing the extraction yield and the loading capacity of the sorbent. The extraction efficiencies of Tyr and HT were up to 61%, with a total solid recovery higher than 90% using an initial concentration of 100 mM biophenol and 10 molar equivalents of ß-CD. The photochemical stability of the complexes thus obtained was estimated from ∆E*ab curve vs. illumination time. The results obtained showed that the phenols encapsulated into solid ß-CD are protected against photodegradation. The powder obtained could be directly developed as a safe-grade food supplement. This simple eco-friendly process could be used for extracting valuable biophenols from olive mill wastewater.

Acylated Anthocyanins from Red Cabbage and Purple Sweet Potato Can Bind Metal Ions and Produce Stable Blue Colors.

Red cabbage (RC) and purple sweet potato (PSP) are naturally rich in acylated cyanidin glycosides that can bind metal ions and develop intramolecular π-stacking interactions between the cyanidin chromophore and the phenolic acyl residues. In this work, a large set of RC and PSP anthocyanins was investigated for its coloring properties in the presence of iron and aluminum ions. Although relatively modest, the structural differences between RC and PSP anthocyanins, i.e., the acylation site at the external glucose of the sophorosyl moiety (C2-OH for RC vs. C6-OH for PSP) and the presence of coordinating acyl groups (caffeoyl) in PSP anthocyanins only, made a large difference in the color expressed by their metal complexes. For instance, the Al3+-induced bathochromic shifts for RC anthocyanins reached ca. 50 nm at pH 6 and pH 7, vs. at best ca. 20 nm for PSP anthocyanins. With Fe2+ (quickly oxidized to Fe3+ in the complexes), the bathochromic shifts for RC anthocyanins were higher, i.e., up to ca. 90 nm at pH 7 and 110 nm at pH 5.7. A kinetic analysis at different metal/ligand molar ratios combined with an investigation by high-resolution mass spectrometry suggested the formation of metal-anthocyanin complexes of 1:1, 1:2, and 1:3 stoichiometries. Contrary to predictions based on steric hindrance, acylation by noncoordinating acyl residues favored metal binding and resulted in complexes having much higher molar absorption coefficients. Moreover, the competition between metal binding and water addition to the free ligands (leading to colorless forms) was less severe, although very dependent on the acylation site(s). Overall, anthocyanins from purple sweet potato, and even more from red cabbage, have a strong potential for development as food colorants expressing red to blue hues depending on pH and metal ion.

Polyphenols bind to low density lipoprotein at biologically relevant concentrations that are protective for heart disease.

There is ample evidence in the epidemiological literature that polyphenols, the major non-vitamin antioxidants in plant foods and beverages, have a beneficial effect on heart disease. Until recently other mechanisms which polyphenols exhibit such as cell signaling and regulating nitric oxide bioavailability have been investigated. The oxidation theory of atherosclerosis implicates LDL oxidation as the beginning step in this process. Nine polyphenols from eight different classes and several of their O-methylether, O-glucuronide and O-sulfate metabolites have been shown in this study to bind to the lipoproteins and protect them from oxidation at lysosomal/inflammatory pH (5.2), and physiological pH (7.4). Polyphenols bind to the apoprotein at pH 7.4 with Kb > 106 M-1 and the number of molecules of polyphenols bound per LDL particle under saturation conditions varied from 0.4 for ferulic acid to 13.1 for quercetin. Competition studies between serum albumin and LDL show that substantial lipoprotein binding occurs even in the presence of a great molar excess of albumin, the major blood protein. These in vitro results are borne out by published human supplementation studies showing that polyphenol metabolites from red wine, olive oil and coffee are found in LDL even after an overnight fast. A single human supplementation with various fruit juices, coffee and tea also produced an ex vivo protection against lipoprotein oxidation under postprandial conditions. This in vivo binding is heart-protective based on published olive oil consumption studies. Relevant to heart disease, we hypothesize that the binding of polyphenols and metabolites to LDL functions as a transport mechanism to carry these antioxidants to the arterial intima, and into endothelial cells and macrophages. Extracellular and intracellular polyphenols and their metabolites are heart-protective by many mechanisms and can also function as potent "intraparticle" and intracellular antioxidants due to their localized concentrations that can reach as high as the micromolar level. Low plasma concentrations make polyphenols and their metabolites poor plasma antioxidants but their concentration in particles such as lipoproteins and cells is high enough for polyphenols to provide cardiovascular protection by direct antioxidant effects and by other mechanisms such as cell signaling.

Binding of Plant Polyphenols to Serum Albumin and LDL: Healthy Implications for Heart Disease.

Cardiovascular disease (CVD) is the leading cause of death in industrialized nations. The initiating event in atherosclerosis, or hardening of the arteries, is oxidation of low density lipoprotein (LDL). Binding with serum albumin and LDL of 41 polyphenols (major antioxidants in plant foods) constituting four classes of flavonoids, three types of phenolic acids, and seven polyphenol conjugate metabolites was investigated indirectly by fluorescence quenching and directly by affinity separation/high-performance liquid chromatography (four of the polyphenols). Stern-Volmer plots yielded K values for the two proteins. Polyphenol binding was significantly stronger for albumin than with LDL. K values were highly correlated with the lipophilicity of the polyphenols. The number of polyphenol molecules determined by quenching was ∼1 for both proteins. Direct analysis under saturation conditions yielded from 2 to 13 molecules of polyphenols/LDL particle. Multiple substituent effects on binding were analyzed. Evidence was put forward that binding of polyphenols to these proteins is protective for CVD by multiple mechanisms.

ß-Cyclodextrin Does not Alter the Bioaccessibility and the Uptake by Caco-2 Cells of Olive By-Product Phenolic Compounds.

Alperujo-a two-phase olive mill waste that is composed of olive vegetation water and solid skin, pulp, and seed fragments - is a highly valuable olive by-product due to its high content in phenolic compounds. In this study, we assessed whether ß-cyclodextrin (ß-CD), which is used to extract and protect alpejuro phenolic compounds (hydroxytyrosol-O-glucoside, tyrosol, caffeic, and p-coumaric acids) could impact on their bioaccessibility (i.e., the percentage of molecule found in the aqueous phase of the digesta) and uptake by intestinal cells, by using an in vitro digestion model and Caco-2 TC7 cells in culture, respectively. Our results showed that ß-CD did not change the bioaccessibility of the selected phenols. Hydroxytyrosol-O-glucoside and caffeic did not cross Caco-2 cell monolayers. Conversely ferulic acid, identified as the main caffeic acid intestinal metabolite, was absorbed through intestinal cell monolayers (~20%). Interestingly, ß-CD moderately but significantly improved the local absorption of tyrosol and p-coumaric acid (2.3 + 1.4% and 8.5 ± 4.2%, respectively, p < 0.05), even if their final bioavailability (expressed as bioaccessibility × absorption by Caco-2 cells) was not modified (16.2 ± 0.6% vs. 16.8 ± 0.5% for tyrosol and 32.0 ± 3.2% vs. 37.2 ± 3.2% for p-coumaric acid, from pure alperujo and alperujo complexed with ß-CD, respectively). Overall, our results show that ß-CD is an interesting extraction and storage agent for phenolic compounds that does not alter their in vitro bioavailability.

Facilitation costs and benefits function simultaneously on stress gradients for animals.

Understanding the variation in species interactions along environmental stress gradients is crucial for making robust ecological predictions about community responses to changing environmental conditions. The facilitation-competition framework has provided a strong basis for predictions (e.g. the stress-gradient hypothesis, SGH), yet the mechanisms behind patterns in animal interactions on stress gradients are poorly explored in particular for mobile animals. Here, we proposed a conceptual framework modelling changes in facilitation costs and benefits along stress gradients and experimentally tested this framework by measuring fitness outcomes of benefactor-beneficiary interactions across resource quality levels. Three arthropod consumer models from a broad array of environmental conditions were used including aquatic detritivores, potato moths and rainforest carrion beetles. We detected a shift to more positive interactions at increasing levels of stress thereby supporting the application of the SGH to mobile animals. While most benefactors paid no significant cost of facilitation, an increase in potato moth beneficiary's growth at high resource stress triggered costs for benefactors. This study is the first to experimentally show that both costs and benefits function simultaneously on stress gradients for animals. The proposed conceptual framework could guide future studies examining species interaction outcomes for both animals and plants in an increasingly stressed world.

Effect of Foods and ß-Cyclodextrin on the Bioaccessibility and the Uptake by Caco-2 Cells of Hydroxytyrosol from Either a Pure Standard or Alperujo.

Hydroxytyrosol bioaccessibility and absorption by the intestinal cells were studied using an in vitro digestion model and Caco-2 TC7 monolayers cells in culture in the presence and absence of ß-cyclodextrin and foods. Hydroxytyrosol was either provided as a pure standard or in an alperujo powder. The presence of foods significantly decreased hydroxytyrosol bioaccessibility and absorption (-20 and -10%, respectively), while ß-cyclodextrin had no effect. Moreover, the presence of other compounds from alperujo in the intestinal compartment reduced hydroxytyrosol absorption by Caco-2 cells compared to pure standard (-60%). The final bioavailability of hydroxytyrosol, defined as its quantity at the basolateral side of cultured cell monolayers compared to the initial amount in the test meal, was 6.9 ± 0.4, 31.1 ± 1.1, and 40.9 ± 1.5% when hydroxytyrosol was from alperujo or a standard administered with or without food, respectively. Our results show that conversely to foods, ß-cyclodextrin does not alter hydroxytyrosol bioavailability.

Involvement of bilitranslocase and beta-glucuronidase in the vascular protection by hydroxytyrosol and its glucuronide metabolites in oxidative stress conditions.

Olive oil vascular benefits have been attributed to hydroxytyrosol (HT). However, HT biological actions are still debated because it is extensively metabolized into glucuronides (GCs). The aim of this study was to test HT and GC vasculoprotective effects and the underlying mechanisms using aorta rings from 8-week-old male Wistar rats. In the absence of oxidative stress, incubation with 100 µM HT or GC for 5 min did not exert any vasorelaxing effect and did not influence the vascular function. Conversely, in condition of oxidative stress [upon incubation with 500 µM tert-butylhydroperoxide (t-BHP) for 30 min], preincubation with HT or GC improved acetylcholine-induced vasorelaxation compared with untreated samples (no t-BHP). This protective effect was lost for GC, but not for HT, when a washing step (15 min) was introduced between preincubation with HT or GC and t-BHP addition, suggesting that only HT enters the cells. In agreement, bilitranslocase inhibition with 100 µM phenylmethanesulfonyl fluoride for 20 min reduced significantly HT, but not GC, effect on the vascular function upon stress induction. Moreover, GC protective effect (improvement of endothelium-dependent relaxation in response to acetylcholine) in oxidative stress conditions was reduced by preincubation of aorta rings with 300 µM D-saccharolactone to inhibit ß-glucuronidase, which can deconjugate polyphenols. Finally, only HT was detected by high-pressure liquid chromatography in aorta rings incubated with GC and t-BHP. These results suggest that, in conditions of oxidative stress, GC can be deconjugated into HT that is transported through the cell membrane by bilitranslocase to protect vascular function.

Effect of Temperature on Acidity and Hydration Equilibrium Constants of Delphinidin-3-O- and Cyanidin-3-O-sambubioside Calculated from Uni- and Multiwavelength Spectroscopic Data.

Delphinidin-3-O-sambubioside and cyanidin-3-O-sambubioside are the main anthocyanins of Hibiscus sabdariffa calyces, traditionally used to make a bright red beverage by decoction in water. At natural pH, these anthocyanins are mainly in their flavylium form (red) in equilibrium with the quinonoid base (purple) and the hemiketal (colorless). For the first time, their acidity and hydration equilibrium constants were obtained from a pH-jump method followed by UV-vis spectroscopy as a function of temperature from 4 to 37 °C. Equilibrium constant determination was also performed by multivariate curve resolution (MCR). Acidity and hydration constants of cyanidin-3-O-sambubioside at 25 °C were 4.12 × 10(-5) and 7.74 × 10(-4), respectively, and were significantly higher for delphinidin-3-O-sambubioside (4.95 × 10(-5) and 1.21 × 10(-3), respectively). MCR enabled the obtaining of concentration and spectrum of each form but led to overestimated values for the equilibrium constants. However, both methods showed that formations of the quinonoid base and hemiketal were endothermic reactions. Equilibrium constants of anthocyanins in the hibiscus extract showed comparable values as for the isolated anthocyanins.

Interactions between Carotenoids from Marine Bacteria and Other Micronutrients: Impact on Stability and Antioxidant Activity.

Recently isolated spore-forming pigmented marine bacteria Bacillus indicus HU36 are sources of oxygenated carotenoids with original structures (about fifteen distinct yellow and orange pigments with acylated d-glucosyl groups). In this study, we evaluated the stability (sensitivity to iron-induced autoxidation) and antioxidant activity (inhibition of iron-induced lipid peroxidation) of combinations of bacterial HU36 carotenoids with the bacterial vitamin menaquinone MQ-7 and with phenolic antioxidants (vitamin E, chlorogenic acid, rutin). Unexpectedly, MQ-7 strongly improves the ability of HU36 carotenoids to inhibit Fe(II)-induced lipid peroxidation, although MQ-7 was not consumed in the medium. We propose that their interaction modifies the carotenoid antioxidant mechanism(s), possibly by allowing carotenoids to scavenge the initiating radicals. For comparison, ß-carotene and lycopene in combination were shown to exhibit a slightly higher stability toward iron-induced autoxidation, as well as an additive antioxidant activity as compared to the carotenoids, individually. HU36 carotenoids and phenolic antioxidants displayed synergistic activities in the inhibition of linoleic acid peroxidation induced by heme iron, but not by free iron. Synergism could arise from antioxidants interacting via electron transfer through the porphyrin nucleus of heme iron. Overall, combining antioxidants acting via complementary mechanisms could be the key for optimizing the activity of this bacterial carotenoid cocktail.

Changes in the distribution of multispecies pest assemblages affect levels of crop damage in warming tropical Andes.

Climate induced species range shifts might create novel interactions among species that may outweigh direct climatic effects. In an agricultural context, climate change might alter the intensity of competition or facilitation interactions among pests with, potentially, negative consequences on the levels of damage to crop. This could threaten the productivity of agricultural systems and have negative impacts on food security, but has yet been poorly considered in studies. In this contribution, we constructed and evaluated process-based species distribution models for three invasive potato pests in the Tropical Andean Region. These three species have been found to co-occur and interact within the same potato tuber, causing different levels of damage to crop. Our models allowed us to predict the current and future distribution of the species and therefore, to assess how damage to crop might change in the future due to novel interactions. In general, our study revealed the main challenges related to distribution modeling of invasive pests in highly heterogeneous regions. It yielded different results for the three species, both in terms of accuracy and distribution, with one species surviving best at lower altitudes and the other two performing better at higher altitudes. As to future distributions our results suggested that the three species will show different responses to climate change, with one of them expanding to higher altitudes, another contracting its range and the other shifting its distribution to higher altitudes. These changes will result in novel areas of co-occurrence and hence, interactions of the pests, which will cause different levels of damage to crop. Combining population dynamics and species distribution models that incorporate interspecific trade-off relationships in different environments revealed a powerful approach to provide predictions about the response of an assemblage of interacting species to future environmental changes and their impact on process rates.

Antioxidant properties of 3-deoxyanthocyanidins and polyphenolic extracts from Côte d'Ivoire's red and white sorghums assessed by ORAC and in vitro LDL oxidisability tests.

Red sorghum is a source of phenolic compounds (PCs), including 3-deoxyanthocyanidins that may protect against oxidative stress related disease such as atherosclerosis. HPLC was used to characterise and quantify PCs extracted from red or white sorghum whole grain flour. Antioxidant activity was measured by an oxygen radical absorbance capacity assay and against LDL-oxidisability, and further compared to that of synthesised 3-deoxyanthocyanidins (i.e., luteolinidin and apigeninidin). Phenolic content of red and white sorghums was evaluated as 3.90 ± 0.01 and 0.07 ± 0.01 mmol gallic acid equivalents L(-1), respectively. Luteolinidin and apigeninidin were mainly found in red sorghum. Red sorghum had almost 3 and 10 times greater specific antioxidant activity compared to luteolinidin and apigeninidin, respectively. Red sorghum PCs and the two 3-deoxyanthocyanidins were also effective at preventing LDL vitamin E depletion and conjugated diene production. Red sorghum flour exhibits antioxidant capacity suggesting that it may be a valuable health-promoting food.

An innovative grape juice enriched in polyphenols by microwave-assisted extraction.

The grape juice by-product obtained from grape traditional press was extracted by Microwave Hydrodiffusion and Gravity (MHG); a green extraction technique preliminarily optimized at 1 W/g. The MHG extract (MHGE) was analyzed by HPLC for identification and quantification of anthocyanins and other phenolic compounds. Then, MHGE was added to the natural juice (NJ) to produce an innovative grape juice (IJ). These three juices were evaluated for their total polyphenol content (TPC), total anthocyanin content (TAC) in addition to their sensorial characteristics. MHGE showed the highest values of TPC (21.41±0.04 mg GAE/g DW), TAC (4.49±0.01 µg MVGE/g DW). Moreover, IJ (grape juice enriched with MHGE) was richer in TPC (6.70±0.0 1mg GAE/g DW) and TAC (3.96±0.01 µg MVGE/g DW) than NJ (2.90±0.02 mg GAE/g DW and 3.63±0.06 µg MVGE/g DW, respectively).

Modeling temperature-dependent survival with small datasets: insights from tropical mountain agricultural pests.

Many regions are increasingly threatened by agricultural pests but suffer from a lack of data that hampers the development of adequate population dynamics models that could contribute to pest management strategies. Here, we present a new model relating pest survival to temperature and compare its performance with two published models. We were particularly interested in their ability to simulate the deleterious effect of extreme temperatures even when adjusted to datasets that did not include extreme temperature conditions. We adjusted the models to survival data of three species of potato tuber moth (PTM), some major pests in the Tropical Andes. To evaluate model performance, we considered both goodness-of-fit and robustness. The latter consisted in evaluating their ability to predict the actual altitudinal limits of the species in the Ecuadorian Andes. We found that even though our model did not always provide the best fit to data, it predicted extreme temperature mortality and altitudinal limits accurately and better than the other two models. Our study shows that the ability to accurately represent the physiological limits of species is important to provide robust predictions of invasive pests' potential distribution, particularly in places where temperatures approach lethal extremes. The value of our model lies in its ability to simulate accurate thermal tolerance curves even with small datasets, which is useful in places where adequate pest management is urgent but data are scarce.

Experimental support of the stress-gradient hypothesis in herbivore-herbivore interactions.

The stress-gradient hypothesis (SGH) postulates an increase in the frequency of positive species interactions at increasing amounts of stress. While the SGH has been extensively tested in plant-plant interactions along abiotic stresses, it remains unclear whether this hypothesis could apply to higher trophic levels, such as herbivores, along biotic stress gradients. To address this issue, we investigated how the interaction between two potato herbivores may change along a stress gradient created by an assortment of potato varieties with different tuber palatability. We used a tuber resistance trait as a measure for biotic stress and one herbivore as the facilitator to gain access to the tuber of the other herbivore. Our experiment revealed a switch from neutral to positive interactions with increasing stress, confirming for the first time the predictions of the SGH for herbivores. Moreover, the intensity of facilitation decreased at high stress levels, suggesting that benefits by the facilitating species were dampened in the most stressful environment. In view of the ubiquitous role played by positive interactions among herbivores, broadening our search image for facilitative effects among other plant enemies will allow a better awareness of the importance of the SGH in structuring plant communities.

Development of a viral biopesticide for the control of the Guatemala potato tuber moth Tecia solanivora.

The Guatemala potato tuber moth Tecia solanivora (Povolny) (Lep. Gelechiidae) is an invasive species from Mesoamerica that has considerably extended its distribution area in recent decades. While this species is considered to be a major potato pest in Venezuela, Colombia, and Ecuador, currently no specific control methods are available for farmers. To address this issue we developed a biopesticide formulation to be used in integrated pest management of T. solanivora, following three steps. First, search for entomopathogenic viruses were carried out through extensive bioprospections in 12 countries worldwide. As a result, new Phthorimaea operculella granulovirus (PhopGV) isolates were found in T. solanivora and five other gelechid species. Second, twenty PhopGV isolates, including both previously known and newly found isolates, were genetically and/or biologically characterized in order to choose the best candidate for a biopesticide formulation. Sequence data were obtained for the ecdysteroid UDP-glucosyltransferase (egt) gene, a single copy gene known to play a role in pathogenicity. Three different sizes (1086, 1305 and 1353 bp) of egt were found among the virus isolates analyzed. Unexpectedly, no obvious correlation between egt size and pathogenicity was found. Bioassays on T. solanivora neonates showed a maximum of a 14-fold difference in pathogenicity among the eight PhopGV isolates tested. The most pathogenic PhopGV isolate, JLZ9f, had a medium lethal concentration (LC(50)) of 10 viral occlusion bodies per square mm of consumed tuber skin. Third, we tested biopesticide dust formulations by mixing a dry carrier (calcium carbonate) with different adjuvants (magnesium chloride or an optical brightener or soya lecithin) and different specific amounts of JLZ9f. During laboratory experiments, satisfactory control of the pest (>98% larva mortality compared to untreated control) was achieved with a formulation containing 10 macerated JLZ9f-dead T. solanivora larvae per kg of calcium carbonate mixed with 50 mL/kg of soya lecithin. The final product provides an interesting alternative to chemical pesticides for Andean farmers affected by this potato pest.

Effects of physicochemical properties of carotenoids on their bioaccessibility, intestinal cell uptake, and blood and tissue concentrations.

SCOPE: Carotenoid bioavailability is affected by numerous factors. Our aim was to assess the involvement of known carotenoid physicochemical properties (e.g., hydrophobicity, van der Waals volume,…) on the transport of the main dietary carotenoids (ß-carotene, lycopene, lutein, and astaxanthin, from their food matrix to their main storage tissues. METHODS AND RESULTS: We used four complementary models: synthetic mixed micelles, an in vitro digestion procedure, Caco-2 cell monolayers, and a gavage experiment in rats. The efficiency with which pure carotenoids were incorporated into synthetic mixed micelles was related to their melting points (r = 0.99, p = 0.015). The efficiency with which pure carotenoids were transferred from dietary triglycerides into mixed micelles was related to carotenoid hydrophobicity (r = -1, p = 0.005). There was no relationship between the carotenoid physicochemical properties studied and their uptake efficiency by Caco-2. The postprandial plasma carotenoid response to carotenoid gavage was related to carotenoid hydrophobicity (r = -0.99, p = 0.006). Carotenoid adipose tissue response was not related to the carotenoid physicochemical properties studied. CONCLUSION: Thus, carotenoid hydrophobicity is important for bioaccessibility and postprandial blood response of carotenoids. In contrast, the carotenoid physicochemical properties studied are apparently not strong determinants of carotenoid uptake by enterocytes and adipose tissue.

Direct enrichment of olive oil in oleuropein by ultrasound-assisted maceration at laboratory and pilot plant scale.

The possibility to improve the nutritional value of olive oil by enriching it in phenolic compounds from olive leaves (e.g., oleuropein) by ultrasonic maceration was studied. The experimental design used led to the following optimal extraction conditions: ultrasonic power of 60 W, temperature of 16°C and sonication duration of 45 min. The high total phenolic content (414.3 ± 3.2mg of oleuropein equivalent/kg of oil), oleuropein (111.0 ± 2.2mg/kg of oil) and α-tocopherol (55.0 ± 2.1g/kg of oil) concentrations obtained by optimized ultrasound-assisted extraction (UAE) proved the efficiency of this process when compared with the conventional solid-liquid extraction. Histochemical analyses showed that this efficiency is due to specific alteration of the phenol-containing leaf structures. Furthermore, the radical-scavenging activity of the processed oil (DPPH test) and its stability toward lipid autoxidation (heating test) confirmed its enrichment in antioxidants. Sensory evaluation of the enriched olive oil showed a slight increase in bitterness but an overall acceptability. Finally, the enriched olive oil was characterized by clear green color (L*, a*, b* parameters).

Chemical synthesis of citrus flavanone glucuronides.

Flavanone glucuronides are the major phenolic metabolites detected in human plasma after consumption of citrus fruits. As such, they might display significant cardioprotective effects. In this work, glucuronides of naringenin (4'- and 7-O-beta-d-glucuronides) and hesperetin (3'- and 7-O-beta-d-glucuronides), the major flavanone aglycones in grapefruit and orange, respectively, have been chemically synthesized. On the one hand, the most reactive hydroxyl group, C7-OH, was protected by selective benzoylation to allow subsequent glucuronidation of C4'-OH (naringenin) or C3'-OH (hesperetin) (B-ring). On the other hand, the selective debenzoylation at C7-OH of the perbenzoylated flavanone aglycones allowed glucuronidation at the same position (A-ring). After careful deprotection, the target compounds were purified and characterized by nuclear magnetic resonance and mass spectrometry.

Microwave-assisted water extraction of green tea polyphenols.

INTRODUCTION: Green tea, a popular drink with beneficial health properties, is a rich source of specific flavanols (polyphenols). There is a special interest in the water extraction of green tea polyphenols since the composition of the corresponding extracts is expected to reflect the one of green tea infusions consumed worldwide. OBJECTIVE: To develop a microwave-assisted water extraction (MWE) of green tea polyphenols. METHODOLOGY: MWE of green tea polyphenols has been investigated as an alternative to water extraction under conventional heating (CWE). The experimental conditions were selected after consideration of both temperature and extraction time. The efficiency and selectivity of the process were determined in terms of extraction time, total phenolic content, chemical composition (HPLC-MS analysis) and antioxidant activity of the extracts. RESULTS: By MWE (80 degrees C, 30 min), the flavanol content of the extract reached 97.46 (+/- 0.08) mg of catechin equivalent/g of green tea extract, vs. only 83.06 (+/- 0.08) by CWE (80 degrees C, 45 min). In particular, the concentration of the most bioactive flavanol EGCG was 77.14 (+/- 0.26) mg of catechin equivalent/g of green tea extract obtained by MWE, vs 64.18 (+/- 0.26) mg/g by CWE. CONCLUSION: MWE appears more efficient than CWE at both 80 and 100 degrees C, particularly for the extraction of flavanols and hydroxycinnamic acids. Although MWE at 100 degrees C typically affords higher yields in total phenols, MWE at 80 degrees C appears more convenient for the extraction of the green tea-specific and chemically sensitive flavanols.