Sylvain Guyot

Université d´Avignon et des Pays du Vaucluse, Avinyó, Provence-Alpes-Côte d'Azur, France

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Publications (64)136.8 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: A histological approach including light microscopy and transmission electron microscopy (TEM) was used to provide accurate information on the localization of condensed tannins in the edible tissues and in the stone of date fruits (Phoenix dactylifera L.). Light microscopy was carried out on fresh tissues after staining by 4-dimethylaminocinnamaldehyde (DMACA) for a specific detection of condensed tannins. Thus, whether under light microscopy or transmission electron microscopy (TEM), results showed that tannins are not located in the epidermis but more deeply in the mesocarp in the vacuole of very large cells. Regarding the stones, tannins are found in a specific cell layer located at 50 μm from the sclereid cells of the testa.
    Journal of agricultural and food chemistry. 07/2014;
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    ABSTRACT: The qualitative and quantitative analysis of phenolic compounds of the methanol extract of Tunisian thornless form Opuntia Ficus Indica flowers was described. Reversed phase high-performance liquid chromatography UV photodiode array analysis detected four peaks at 320 nm, attributed to phenolic acids; three of them were quantified to 141−1 of fresh plant material. Electrospray ionization mass spectrometer analysis detected 8 glycosylated flavonols at 350 nm, identified among 11 chromatographic peaks. The amount of flavonoids was 4980−1 of fresh plant material, with isorhamnetin glycosylated derivatives as the main flavonoid components (55.1%), quercetin derivatives as the second (23.7%), and kaempferol derivatives as the third (8.4%).
    International Journal of Food Properties 01/2014; 17(4). · 0.88 Impact Factor
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    ABSTRACT: Polyphenols have a favorable antioxidant potential on human health, suggesting that their high content in apple is responsible for the beneficial effects of apple consumption. They are also linked to the quality of apple juices and ciders since they are predominantly responsible for astringency, bitterness, color and aroma. Major phenolic compounds were quantified by liquid chromatography in fruits and juices from a cider apple progeny harvested for three years. The total content of procyanidins and their average degree of polymerization (DPn) were also determined in fruits by phloroglucinolysis. Variability and extraction yield of these compounds were determined. The variability observed in the progeny was representative of the variability observed in many cider apple varieties. Hydroxycinnamic acids were the most extractable group, with an average extraction yield of 67%, whereas flavonols and anthocyanins were the least. This study is the first one to introduce variability and extraction yields of the main phenolic compounds in both fruits and juices of a cider apple progeny. This dataset will be used for an upcoming QTL mapping study, an original approach that has never been undertaken for cider apple.
    Journal of the Science of Food and Agriculture 10/2013; · 1.76 Impact Factor
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    ABSTRACT: Flavonoids, like other metabolites synthesized via the phenylpropanoid pathway, possess a wide range of biological activities including functions in plant development and its interaction with the environment. Dihydrochalcones (mainly phloridzin, sieboldin, trilobatin, phloretin) represent the major flavonoid subgroup in apple green tissues. Although this class of phenolic compounds is found in very large amounts in some tissues (≈200mg/g of leaf DW), their physiological significance remains unclear. In the present study, we highlight their tissue-specific localization in young growing shoots suggesting a specific role in important physiological processes, most notably in response to biotic stress. Indeed, dihydrochalcones could constitute a basal defense, in particular phloretin which exhibits a strong broad-range bactericidal and fungicidal activity. Our results also indicate that sieboldin forms complexes with iron with strong affinity, reinforcing its antioxidant properties and conferring to this dihydrochalcone a potential for iron seclusion and/or storage. The importance of localization and biochemical properties of dihydrochalcones are discussed in view of the apple tree defense strategy against both biotic and abiotic stresses.
    Phytochemistry 04/2013; · 3.05 Impact Factor
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    ABSTRACT: The necrogenic bacterium Erwinia amylovora responsible for the fire blight disease causes cell death in apple tissues to enrich intercellular spaces with nutrients. Apple leaves contain large amounts of dihydrochalcones (DHCs), including phloridzin and its aglycone phloretin. Previous work showed an important decrease in the constitutive DHCs stock in infected leaves, probably caused by transformation reactions during the infection process. At least two flavonoid transformation pathways have been described so far: deglucosylation and oxidation. The aim of the present study was to determine whether DHCs are differentially converted in two apple genotypes displaying contrasted susceptibilities to the disease. Different analyses were performed: i) enzymatic activity assays in infected leaves, ii) identification/quantification of end-products obtained after in vitro enzymatic reactions with DHCs, iii) evaluation of the bactericidal activity of end-products. The results of the enzymatic assays showed that deglucosylation was dominant over oxidation in the susceptible genotype MM106 while the opposite was observed in the resistant genotype Evereste. These data were confirmed by LC-UV/Vis-MS analysis of in vitro reaction mixtures, especially because higher levels of o-quinoid oxidation products of phloretin were measured by using the enzymatic extracts of Evereste infected leaves. Their presence correlated well with a strong bactericidal activity of the reaction mixtures. Thus, our results suggest that a differential transformation of DHCs occur in apple genotypes with a potential involvement in the establishment of the susceptibility or the resistance to fire blight, through the release of glucose or of highly bactericidal compounds respectively.
    Plant Physiology and Biochemistry 03/2013; · 2.78 Impact Factor
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    ABSTRACT: The polyphenol profile of two Tunisian varieties of dates including flavanols, flavonols, flavones and hydroxycinnamates was characterized. Three tissue zones (flesh, peel and stone) and three maturity stages were considered. Phenolic compounds were analyzed using reversed phase high performance liquid chromatography coupled to UV-visible and electrospray mass spectrometry. Extractable procyanidin oligomers and non-extractable polymers were characterized and quantified using phloroglucinolysis prior to HPLC analysis. Non-extractable procyanidin polymers based on (-)-epicatechin structure were by far the most concentrated polyphenols in ripe dates, accounting for 95% of total polyphenols with an average concentration of 14 g/kg in the fresh edible parts of the fruit. Interestingly, procyanidins were also highly concentrated in the stones. The concentration and average degree of polymerization (DPn) of the procyanidins decreased according to maturity. Other phenolics, including caffeoylshikimic acid hexoside, cafeoyl-sinapoyl monohexoside and dihexoside, and acetylated flavonols, were tentatively identified for the first time in the fruit.
    Journal of Agricultural and Food Chemistry 02/2013; · 3.11 Impact Factor
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    ABSTRACT: This study evaluated the phenols of the Gala, Fuji and Golden Delicious varieties, which make up 95% of Brazilian production. The phenolic profiles (whole fruit) were determined by high pressure liquid chromatography, total phenols were determined using the Folin-Ciocalteau method and antioxidant capacity by the FRAP method (whole fruit, skin, pulp and centre). The Golden Delicious had the highest phenol content (408 mg/fruit) compared to Fuji (194 mg/fruit) and Gala (162 mg/fruit), and the antioxidant capacity of the Golden Delicious was 2.5 and 3.6 times higher than that found in the Fuji and Gala, respectively. The phenolic profile for the three varieties showed 5-caffeoylquinic acid, ( - )-epicatechin, procyanidin B2 and phloridzin as major components with procyanidins as the predominant class and quercetin as the minority. The consumption of apple pulp may provide 48-78% of the antioxidant capacity; however, if the peel is ingested, it may increase to 79-89%. The type of apple variety and the parts ingested are factors that must be taken into consideration in consumption.
    International Journal of Food Sciences and Nutrition 01/2013; · 1.26 Impact Factor
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    ABSTRACT: Polyphenols have favorable antioxidant potential on human health suggesting that their high content in apple is responsible for the beneficial effects of apple consumption. They are also related to the quality of ciders as they predominantly account for astringency, bitterness, color and aroma. Five groups of phenolic compounds are described in the apple fruit: flavanols, hydroxycinnamic acids, dihydrochalcones, flavonols and anthocyanins. So far, only two studies have been published on the genetic basis of the phenolic content of dessert apples. As cider apples are commonly described to be much more concentrated in phenolic compounds than dessert varieties, the present study focuses on a cider apple progeny. 32 compounds belonging to the five groups were identified and quantified by HPLC-UV and UHPLC-UV-MS/MS in fruit extracts and juices. 53 QTL controlling phenolic compounds concentration were detected on nine linkage groups (LG) on the integrated linkage map, for all phenolic groups except anthocyanins. QTL clusters located on LG1, 12, 14, 15 and 17 were stable across the year or the studied material. QTL detected on LG1, 14 and 17 for quercitrin, p-coumaroylquinic acid, rutin and chlorogenic acid confirmed results of previous studies. However, no significant QTL was obtained on the LG16 where a major locus for flavanols was previously located. With the two previous studies, this study shows the diversity of genomic regions controlling traits of interest in apple.
    Sixth Rosaceous Genomics Conference, Mezzocorona, Italy; 10/2012
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    ABSTRACT: PURPOSE: Syrah red grapes are used in the production of tannin-rich red wines. Tannins are high molecular weight molecules, proanthocyanidins (PAs), and poorly absorbed in the upper intestine. In this study, gut microbial metabolism of Syrah grape phenolic compounds was investigated. METHODS: Syrah grape pericarp was subjected to an enzymatic in vitro digestion model, and red wine and grape skin PA fraction were prepared. Microbial conversion was screened using an in vitro colon model with faecal microbiota, by measurement of short-chain fatty acids by gas chromatography (GC) and microbial phenolic metabolites using GC with mass detection (GC-MS). Red wine metabolites were further profiled using two-dimensional GC mass spectrometry (GCxGC-TOFMS). In addition, the effect of PA structure and dose on conversion efficiency was investigated by GC-MS. RESULTS: Red wine exhibited a higher degree of C1-C3 phenolic acid formation than PA fraction or grape pericarp powders. Hydroxyphenyl valeric acid (flavanols and PAs as precursors) and 3,5-dimethoxy-4-hydroxybenzoic acid (anthocyanin as a precursor) were identified from the red wine metabolite profile. In the absence of native grape pericarp or red wine matrix, the isolated PAs were found to be effective in the dose-dependent inhibition of microbial conversions and short-chain fatty acid formation. CONCLUSIONS: Metabolite profiling was complementary to targeted analysis. The identified metabolites had biological relevance, because the structures of the metabolites resembled fragments of their grape phenolic precursors or were in agreement with literature data.
    European Journal of Nutrition 06/2012; · 3.13 Impact Factor
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    ABSTRACT: The complexation kinetics of β-casein with tannins were investigated by means of stopped flow and small-angle X-ray scattering (SAXS). Several small plant tannins have been considered: epigallocatechin-gallate (EGCG) from green tea and a set of oligomeric tannins from apples. We show that the kinetics are composed of two processes. The first process is a rapid uptake of tannins by the β-casein micelles over 40–100 ms and the second process is a slow reorganization of the tannin-dressed proteins into stable heavier micelles over a period of up to 200 s. In the first process, the protein segments in the cores of the micelles are rapidly coated by tannins. Detailed analysis of the SAXS profiles during the slow dynamics reveals that the system remains composed of micelles whose structural attributes evolve smoothly toward equilibrium values. The quantity of the bound tannins remains constant during the whole slow evolution of the system. We conclude that the dominant elementary events that drive the slow kinetics are the exchange processes of tannin-dressed proteins from one micelle to another.
    RSC Advances 04/2012; 2(9):3934-3941. · 3.71 Impact Factor
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    ABSTRACT: Interactions of plant tannins with polysaccharide hyaluronan are studied by means of light scattering and small-angle X-ray scattering (SAXS). In this paper, we show that (1) the tannin-polysaccharide complexes remain stable in colloidal suspension; (2) the masses and structures of colloidal tannin-polysaccharide objects depend on the tannin degree of polymerization; and (3) the densities of tannin-polysaccharide aggregates are about 7 times lower than the density of a single solvated polysaccharide molecule. Short tannins and polysaccharides are aggregated in loose oligomeric structures whose sizes are comparable to a single polysaccharide molecule. Tannins longer than 10 nm and polysaccharides are aggregated in larger microgel-like particles whose sizes exceed 200 nm.
    Biomacromolecules 02/2012; 13(3):751-9. · 5.37 Impact Factor
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    ABSTRACT: Apple (Malus×domestica) leaf phenolics, in particular the three major dihydrochalcones, phloridzin, trilobatin and sieboldin, were studied for their potential contribution to the resistance to Erwinia amylovora, the causative agent of fire blight. The constitutive phenolic compositions of part of a progeny resulting from a cross between two apple genotypes, displaying contrasted susceptibilities to fire blight, were investigated by RP–HPLC–DAD. With regards to the major dihydrochalcones, offsprings were found to display the parent profile: either phloridzin alone or a combination of phloridzin, trilobatin and sieboldin. Despite a strong antioxidant activity that could interfere with the oxidative burst triggered by the bacteria, sieboldin did not segregate with resistance even though it was strongly associated with a high constitutive antioxidant capacity of leaf extracts. Several hydroxycinnamic acids and flavonols were identified in the progeny but profiles were very similar. However, concentrations of several flavonols positively correlated with resistance in the genotypes that possessed trilobatin and sieboldin. The major dihydrochalcones that represent more than 200mg/g of leaf DW are not directly responsible for resistance to fire blight in this system. In addition, a high constitutive antioxidant activity is unlikely to be sufficient to block bacterial spread.
    Environmental and Experimental Botany 12/2011; 74:65-73. · 3.00 Impact Factor
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    ABSTRACT: Procyanidins (i.e. condensed tannins) are polyphenols commonly found in fruits. During juice and cider making, apple polyphenol oxidase catalyzes the oxidation of caffeoylquinic acid (CQA) into its corresponding o-quinone which further reacts with procyanidins and other polyphenols, leading to the formation of numerous oxidation products. However, the structure and the reaction pathways of these neoformed phenolic compounds are still largely unknown. Experiments were carried out on a model system to gain insights into the chemical processes occurring during the initial steps of fruit processing. Procyanidin B2 was oxidized by caffeoylquinic acid o-quinone (CQAoq) in an apple juice model solution. The reaction products were monitored using high performance liquid chromatography (HPLC) coupled to ultraviolet (UV)-visible and electrospray tandem mass spectrometry (ESI-MS/MS) in the negative mode. Oxidative conversion of procyanidin B2 ([M-H](-) at m/z 577) into procyanidin A2 at m/z 575 was unambiguously confirmed. In addition, several classes of products were characterized by their deprotonated molecules ([M-H](-)) and their MS/MS fragmentation patterns: hetero-dimers (m/z 929) and homo-dimers (m/z 1153 and 705) resulting from dimerization involving procyanidin and CQA molecules; intramolecular addition products at m/z 575, 573, 927, 1151 and 703. Interestingly, no extensive polymerization was observed. Analysis of a cider apple juice enabled comparison with the results obtained on a biosynthetic model solution. However, procyanidin A2 did not accumulate but seemed to be an intermediate in the formation of an end-product at m/z 573 for which two structural hypotheses are given. These structural modifications of native polyphenols as a consequence of oxidation probably have an impact on the organoleptic and nutritional properties of apple juices and other apple-derived foods.
    Biological Mass Spectrometry 11/2011; 46(11):1186-97. · 3.41 Impact Factor
  • Sylvain Guyot, Pascal Poupard
    05/2011: pages 103-158; , ISBN: 2743013389, 9782743013387
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    ABSTRACT: Proanthocyanidins (PAs) are seed coat flavonoids that impair the digestibility of Brassica napus meal. Development of low-PA lines is associated with a high-quality meal and with increased contents in oil and proteins, but requires better knowledge of seed flavonoids. Flavonoids in Brassica mature seed are mostly insoluble so that very few qualitative and quantitative data are available yet. In the present study, the profiling of seed coat flavonoids was established in eight black-seeded B. napus genotypes, during seed development when soluble flavonoids were present and predominated over the insoluble forms. Thirteen different flavonoids including (-)-epicatechin, five procyanidins (PCs which are PAs composed of epicatechin oligomers only) and seven flavonols (quercetin-3-O-glucoside, quercetin-dihexoside, isorhamnetin-3-O-glucoside, isorhamnetin-hexoside-sulfate, isorhamnetin-dihexoside, isorhamnetin-sinapoyl-trihexoside and kaempferol-sinapoyl-trihexoside) were identified and quantified using liquid chromatography coupled to electrospray ionization-mass spectrometry (LC-ESI-MS(n)). These flavonol derivatives were characterized for the first time in the seed coat of B. napus, and isorhamnetin-hexoside-sulfate and isorhamnetin-sinapoyl-trihexoside were newly identified in Brassica spp. High amounts of PCs accumulated in the seed coat, with solvent-soluble polymers of (-)-epicatechin reaching up to 10% of the seed coat weight during seed maturation. In addition, variability for both PC and flavonol contents was observed within the panel of eight black-seeded genotypes. Our results provide new insights into breeding for low-PC B. napus genotypes.
    Journal of Agricultural and Food Chemistry 05/2010; 58(10):6246-56. · 3.11 Impact Factor
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    ABSTRACT: Since the gut microbiota metabolizes various dietary constituents unabsorbed by the small intestine and modulates colon function, it plays an essential role in colon carcinogenesis. First, we have developed a model of human microbiota-associated rats (HMA), fed a human-type diet and injected with 1-2,dimethylhydrazine (DMH). We observed that the number and size of DMH-induced aberrant crypt foci (ACF) were significantly higher in HMA rats than in germ-free or conventional rats. Second, we used this model to assess the protective effect of an apple proanthocyanidin-rich extract (APE) on colon carcinogenesis. In this model, ACF number and multiplicity were not reduced by APE at 0.001% and 0.01% in drinking water. They were higher with APE 0.1% than with APE 0.01%. Therefore, the cross-talk between human microbiota and the colon epithelium should be taken into account in carcinogenesis models. Moreover, attention should be paid prior to using proanthocyanidin extracts as dietary supplements for humans.
    Journal of Agricultural and Food Chemistry 03/2010; 58(7):4120-5. · 3.11 Impact Factor
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    ABSTRACT: Procyanidins commonly known as condensed tannins are a type of polyphenol with wide abundance naturally. They are commonly known as potent anti-oxidants with powerful free radical scavenging activity as well as anti-tumor-promoting activity. Little is known about the enzymatic mechanisms/pathways involved in the microbial biotransformation of these polyphenolic molecules. The extracellular enzyme, dioxygenase produced by Aspergillus fumigatus was used as in vitro tools to study the degradation pathway of a model procyanidin dimer, namely procyanidin B2. The enzyme was purified to homogeneity by a two step process of anion-exchange chromatography coupled with FPLC followed by gel-filtration chromatography coupled with HPLC and the molecular mass estimated. In addition, the different biotransformed products resulted from the dioxygenase action on PB2 were purified using Reversed-Phase-High Performance Liquid Chromatography prior to their identification and characterization by structural elucidation using Electrospray Ionization-Mass Spectrometry. Subsequently, the mechanism of dioxygenase action on procyanidin dimer was defined.
    Process Biochemistry. 02/2010; 45(6):904-913.
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    ABSTRACT: A regular consumption of fresh or processed fruits can contribute to preventing cardiovascular diseases and cancer. Polyphenols, which are naturally present in fresh apples (Malus x domestica), are widely acknowledged as antioxidants. Cider apples contain high concentrations of polyphenols, but their composition differs markedly as a function of cultivar. Two of which were studied in this respect. The first cultivar was Marie Menard, which is highly concentrated in polyphenols and has a standard profile showing high levels of hydroxycinnamic acids and monomeric catechins, widely involved in oxidation reactions catalyzed by the polyphenol oxidase; the second one was Guillevic, which has an atypical phenolic profile for a cider apple, with highly polymerized procyanidins and no monomeric catechins, which thus induces a low sensitivity to enzymatic browning. Both cultivars were subjected to osmotic dehydration at two temperatures (45 and 60 °C) or to convective air-drying, and a combination of the two processes was also tested. Phenolic compounds were quantified by high performance liquid chromatography for various processing times and regardless of the quantity of impregnated sugar. The results revealed different behaviors depending on the polyphenolic groups of compounds. Procyanidins were better preserved by the processes than hydroxycinnamic acids or monomeric catechins. Indeed, these latter groups of polyphenols were first of all involved in enzymatic browning but could also diffuse more easily as their molecular weight was lower. Polyphenol retention was also dependent on the process applied. Polyphenols were better retained by convective drying than by osmotic dehydration, and when soaking was applied as a pretreatment, polyphenol losses were limited during subsequent drying. Moreover, the level of sucrose impregnation could mask the astringency of procyanidins. KeywordsDehydration-Impregnation-Diffusion-Oxidation-Hydroxycinnamic acids-Procyanidins
    Food and Bioprocess Technology 01/2010; 3(6):867-877. · 4.12 Impact Factor
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    ABSTRACT: Flavonoids are a group of polyphenol compounds with known antioxidant activities. Among them, dihydrochalcones are mainly found in apple leaves (Malus domestica). Glycosylated dihydrochalcones were previously found in large amounts in leaves of two genotypes of Malus with contrasting resistance to fire blight, a bacterial disease caused by Erwinia amylovora. In the present study we demonstrate that soluble polyphenol patterns comprised phloridzin alone or in combination with two additional dihydrochalcones, identified as sieboldin and trilobatin. Presence of sieboldin in young leaves correlated well with a high 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity. Moreover, these leaves displayed enhanced tolerance to paraquat, a photooxidative-stress generating herbicide. Interestingly, phloridzin had a high activity in the oxygen radical absorbance capacity (ORAC) assay, but its presence alone in leaves did not correlate with tolerance to paraquat. In order to further characterise the activity of these compounds, we tested their ability to prevent oxidative-dependent formation of advanced glycation end-products (AGEs) and phenylephrine-induced contraction of isolated rat mesenteric arteries. The antioxidant capacity of sieboldin was clearly demonstrated by showing that this compound (i) prevented vasoconstriction and (ii) inhibited AGEs formation. Both assays provided interesting information concerning a potential use of sieboldin as a therapeutic. Hence, our results strongly argue for a bioactivity of dihydrochalcones as functional antioxidants in the resistance of Malus leaves to oxidative stress. In addition, we demonstrate for the first time that sieboldin is a powerful multipotent antioxidant, effective in preventing physiopathological processes. Further work should aim at demonstrating the potential use of this compound as a therapeutic in treating free radical-involving diseases.
    Phytochemistry 12/2009; 71(4):443-52. · 3.05 Impact Factor
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    ABSTRACT: Several cultivars of apples (Malus domestica) were chosen for their variable concentrations and compositions in phenolic compounds. Cubed samples (1 cm3) were subjected to osmotic dehydration, and the effect of temperature was studied at 45 and 60 degrees C. Water loss, sucrose impregnation, and the evolution of some natural components of the product were followed to quantify mass transfer. Ascorbic acid and polyphenols were quantified by HPLC for several osmotic dehydration times and regardless of the quantity of impregnated sugar. Changes in antioxidant components differed as a function of the nature of molecules. Their concentrations decreased in line with temperature, and few differences were observed between cultivars. Processing at a lower temperature (45 degrees C) caused a total loss in ascorbic acid but allowed the retention of between 74 and 85% of initial polyphenols, depending on the cultivar. Cultivars containing highly polymerized procyanidins (such as Guillevic) experienced less loss. Hydroxycinnamic acids and monomeric catechins displayed the most marked changes. Leaching with water into the soaking solution was the principal mechanism retained to explain these losses.
    Journal of Agricultural and Food Chemistry 12/2009; 58(1):606-14. · 3.11 Impact Factor

Publication Stats

1k Citations
136.80 Total Impact Points


  • 2009
    • Université d´Avignon et des Pays du Vaucluse
      Avinyó, Provence-Alpes-Côte d'Azur, France
    • University of Angers
      Angers, Pays de la Loire, France
    • University of Manitoba
      Winnipeg, Manitoba, Canada
  • 2002–2008
    • University of Aveiro
      • Department of Chemistry
      Aveiro, Aveiro, Portugal
  • 2001–2008
    • French National Institute for Agricultural Research
      Lutetia Parisorum, Île-de-France, France
  • 2007
    • Université Paul Cézanne
      Aix, Provence-Alpes-Côte d'Azur, France
  • 2005–2006
    • Instituto Politécnico de Bragança
      Браганца, Bragança, Portugal
  • 2004
    • Universite des Sciences et Techniques de Masuku
      Franceville, Haut-Ogooué, Gabon
    • Institute of Research for Development
      Marsiglia, Provence-Alpes-Côte d'Azur, France