Reactivity of flavanols: Their fate in physical food processing and recent advances in their analysis by depolymerization.

Flavanols, a subgroup of polyphenols, are secondary metabolites with antioxidant properties naturally produced in various plants (e.g., green tea, cocoa, grapes, and apples); they are a major polyphenol class in human foods and beverages, and have recognized effect on maintaining human health. Therefore, it is necessary to evaluate their changes (i.e., oxidation, polymerization, degradation, and epimerization) during various physical processing (i.e., heating, drying, mechanical shearing, high-pressure, ultrasound, and radiation) to improve the nutritional value of food products. However, the roles of flavanols, in particular for their polymerized forms, are often underestimated, for a large part because of analytical challenges: they are difficult to extract quantitatively, and their quantification demands chemical reactions. This review examines the existing data on the effects of different physical processing techniques on the content of flavanols and highlights the changes in epimerization and degree of polymerization, as well as some of the latest acidolysis methods for proanthocyanidin characterization and quantification. More and more evidence show that physical processing can affect content but also modify the structure of flavanols by promoting a series of internal reactions. The most important reactivity of flavanols in processing includes oxidative coupling and rearrangements, chain cleavage, structural rearrangements (e.g., polymerization, degradation, and epimerization), and addition to other macromolecules, that is, proteins and polysaccharides. Some acidolysis methods for the analysis of polymeric proanthocyanidins have been updated, which has contributed to complete analysis of proanthocyanidin structures in particular regarding their proportion of A-type proanthocyanidins and their degree of polymerization in various plants. However, future research is also needed to better extract and characterize high-polymer proanthocyanidins, whether in their native or modified forms.

Tannin-controlled micelles and fibrils of κ-casein.

Effects of green tea tannin epigallocatechin-gallate (EGCG) on thermal-stress-induced amyloid fibril formation of reduced carboxymethylated bovine milk protein κ-casein were studied by dynamical light scattering and small angle X-ray scattering (SAXS). Two populations of aggregates, micelles, and fibrils dominated the time evolution of light scattering intensity and of effective hydrodynamic diameter. SAXS experiments allowed us to resolve micelles and fibrils so that the time dependence of the scattering profile revealed the structural evolution of the two populations. The low-Q scattering intensity prior to an expected increase in time due to fibril growth shows an intriguing rapid decrease, which is interpreted as the release of monomers from micelles. This phenomenon, observed both in the absence and in the presence of EGCG, indicates that under thermal stress free conditions, native monomers are converted to amyloid-prone monomers that do not form micelles. The consumption of free native monomers results in a release of native monomers from micelles because only native proteins participate in micelle-monomer (quasi)equilibrium. This release is reversible, indicating also that native-to-amyloid-prone monomer conversion is reversible as well. We show that EGCG does not bind to protein in fibrils, neither does it affect/prevent the proamyloid conversion of monomers. EGCG hinders the addition of monomers to growing fibrils. These facts allowed us to propose the kinetics model for EGCG-controlled amyloid aggregation of micellar proteins. Therein, we introduced the growth-rate inhibition function, which quantitatively accounts for the effect of EGCG on the fibril growth at any degree of thermal stress.

Historical biogeography of Reticulitermes termites (Isoptera: Rhinotermitidae) inferred from analyses of mitochondrial and nuclear loci.

Termites of the genus Reticulitermes are ecologically and economically important wood-feeding social insects that are widespread in the Holarctic region. Despite their importance, no study has yet attempted to reconstruct a global time-scaled phylogeny of Reticulitermes termites. In this study, we sequenced mitochondrial (2096bp) and nuclear (829bp) loci from 61 Reticulitermes specimens, collected across the genus' entire range, and one specimen of Coptotermes formosanus, which served as an outgroup. Bayesian and Maximum likelihood analyses conducted on the mitochondrial and nuclear sequences support the existence of four main lineages that span four global geographical regions: North America (NA lineage), western Europe (WE lineage), a region including eastern Europe and western Asia (EA+WA lineage), and eastern Asia (EA lineage). The mitochondrial data allowed us to clarify the phylogenetic relationships among these lineages. They were also used to infer a chronogram that was time scaled based on age estimates for termite fossils (including the oldest Reticulitermes fossils, which date back to the late Eocene-early Oligocene). Our results support the hypothesis that the extant Reticulitermes lineage first differentiated in North America. The first divergence event in the ancestral lineage of Reticulitermes occurred in the early Miocene and separated the Nearctic lineages (i.e., the NA lineages) from the Palearctic lineages (i.e., WE, EE+WA, and EA lineages). Our analyses revealed that the main lineages of Reticulitermes diversified because of vicariance and migration events, which were probably induced by major paleogeographic and paleoclimatic changes that occurred during the Cenozoic era. This is the first global and comprehensive phylogenetic study of Reticulitermes termites, and it provides a crucial foundation for studying the evolution of phenotypic and life-history traits in Reticulitermes. For instance, the phylogeny we obtained suggested that 'asexual queen succession', a unique reproductive system, independently evolved at least three times during the diversification of the genus.

Polyphenol variability in the fruits and juices of a cider apple progeny.

BACKGROUND: Polyphenols have a favourable 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 and colour. Major phenolic compounds were quantified by liquid chromatography in fruits and juices from a cider apple progeny harvested for 3 years. The total content of procyanidins and their average degree of polymerisation (DPn) were also determined in fruits by phloroglucinolysis. Variability and extraction yield of these compounds were determined. RESULTS: 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. CONCLUSION: This study is the first 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.

Interactions between Salivary Proteins and Apple Polyphenols and the Fate of Complexes during Gastric Digestion.

Beneficial polyphenols in apples can reach the stomach as complexes formed with salivary proteins. The present study aimed at documenting the interactions between salivary proteins and cider apple polyphenols and the fate of complexes during gastric digestion. A polyphenolic extract was mixed with human saliva, and interactions were characterized by analyzing proteins and polyphenols in the insoluble and soluble fractions of the mixtures, before and after in vitro gastric digestion. Results confirmed that proline-rich proteins can efficiently precipitate polyphenols and suggested that two zinc-binding proteins can also form insoluble complexes with polyphenols. The classes of polyphenols involved in such complexes depended on the polyphenol-to-protein ratio. In vitro gastric digestion led to extensive proteolysis of salivary proteins, and we formulate the hypothesis that the resulting peptides can interact with and precipitate some procyanidins. Saliva may therefore partly modulate the bioaccessibility of at least procyanidins in the gastric compartment.

Characterization of microbial metabolism of Syrah grape products in an in vitro colon model using targeted and non-targeted analytical approaches.

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.

Phenolic profile and antioxidant capacity of the principal apples produced in Brazil.

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.

Physical and Oxidative Stabilization of Oil-In-Water Emulsions by Roasted Coffee Fractions: Interface- and Continuous Phase-Related Effects.

Emulsions fortified with polyunsaturated fatty acids are highly relevant from a nutritional perspective; however, such products are prone to lipid oxidation. In the current work, this is mitigated by the use of natural antioxidants occurring in coffee. Coffee fractions with different molecular weights were extracted from roasted coffee beans. These components were positioned either at the interface or in the continuous phase of emulsions where they contributed to emulsion stability via different pathways. Coffee brew as a whole, and its high-molecular-weight fraction (HMWF), was able to form emulsions with good physical stability and excellent oxidative stability. When added post-homogenization to the continuous phase of dairy protein-stabilized emulsions, all coffee fractions were able to slow down lipid oxidation considerably without altering the physical stability of emulsions, though HMWF was more effective in retarding lipid oxidation than whole coffee brew or low-molecular-weight fraction. This is caused by various effects, such as the antioxidant properties of coffee extracts, the partitioning of components in the emulsions, and the nature of the phenolic compounds. Our research shows that coffee extracts can be used effectively as multifunctional stabilizers in dispersed systems leading to emulsion products with high chemical and physical stability.

Oil Bodies from Chia (Salvia hispanica L.) and Camelina (Camelina sativa L.) Seeds for Innovative Food Applications: Microstructure, Composition and Physical Stability.

Exploring and deciphering the biodiversity of oil bodies (OBs) recovered from oilseeds are of growing interest in the preparation of sustainable, natural and healthy plant-based food products. This study focused on chia (Salvia hispanica L.) and camelina (Camelina sativa L.) seed OBs. A green refinery process including ultrasound to remove mucilage, aqueous extraction by grinding and centrifugation to recover OBs from the seeds was used. The microstructure, composition and physical stability of the OBs were examined. Confocal laser scanning microscopy images showed that chia and camelina seed OBs are spherical assemblies coated by a layer of phospholipids and proteins, which have been identified by gel electrophoresis. The mean diameters determined by laser light scattering measurements were 2.3 and 1.6 µm for chia and camelina seed OBs, respectively. The chia and camelina seed OBs were rich in lipids and other bioactive components with, respectively, 64% and 30% α-linolenic acid representing 70% and 53% of the total fatty acids in the sn-2 position of the triacylglycerols, 0.23% and 0.26% phospholipids, 3069 and 2674 mg/kg oil of ß-sitosterol, and lipophilic antioxidants: 400 and 670 mg/kg oil of γ-tocopherol. Phenolic compounds were recovered from the aqueous extracts, such as rutin from camelina and caffeic acid from chia. Zeta-potential measurements showed changes from about -40 mV (pH 9) to values that were positive below the isoelectric points of pH 5.1 and 3.6 for chia and camelina seed OBs, respectively. Below pH 6.5, physical instability of the natural oil-in-water emulsions with aggregation and phase separation was found. This study will contribute to the development of innovative and sustainable food products based on natural oil-in-water emulsions containing chia and camelina seed OBs for their nutritional and health benefits.

Effect of some local plant extracts on fatty acid composition of fish (Alestes baremoze) during smoking and sun drying in the Far-North region of Cameroon.

The objective of this study was to assess the antioxidant activities of three plant extracts (Moringa oleifera leaves, Xylopia aethiopica fruits, and Allium cepa leaves) and to evaluate their effects on the preservation of fish polyunsaturated fatty acids (PUFAs) during smoking and sun-drying processes. PUFAs are highly prone to oxidation during fish processing. The plant extracts were analyzed for their polyphenol contents and were evaluated for their total antiradical capacity. The polyphenol components of each plant were characterized. The hydroethanolic and aqueous extracts were added to the fish at concentrations of 3, 6, 9, and 12 g/L and 10, 20, 30, and 40 g/L, respectively. Butylated hydroxytoluene (BHT) was used as a positive control at a concentration of 2 g/L to compare the antioxidant effects of the plant extracts. The treated fish was subjected to smoking or sun drying and the fatty acid composition of the fish lipid extract was assessed. The results showed that the total polyphenolic, flavonoid, and tannin contents varied significantly from one plant extract to the other (p < .05). The radical scavenging and FRAP increased significantly with the concentration of the plant extracts (p < .05). An HPLC analysis of the extracts led to the preliminary identification of four hydroxycinnamic acids in M. oleifera and X. aethiopica, one anthocyanin and one flavone glycoside in M. oleifera, and four flavan-3-ols in X. aethiopica. Moreover, eight flavonols were preliminarily identified in the three plants. Compared to the control product, these plant extracts significantly protected fish PUFAs from oxidation (p < .05). The aqueous extract of A. cepa at 40 g/L better preserved omega-3 in fish during smoking and sun drying than the control product. Incorporating the three plant extracts during smoking and sun-drying processes can effectively preserve the PUFAs in fish. Therefore, these plants are viable sources of natural antioxidants in the preservation of fish products.

Structural properties of colloidal complexes between condensed tannins and polysaccharide hyaluronan.

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.

Interactions between Salivary Proteins and Dietary Polyphenols: Potential Consequences on Gastrointestinal Digestive Events.

The present review documents the current knowledge and hypotheses on how polyphenols-saliva interactions may modulate the bioaccessibility or bioavailability of nutrients and highlights research prospects in the field. After an updated description of the different classes of dietary polyphenols and their modifications by food processing or digestion, an overview of interactions between salivary proteins and polyphenols (with an emphasis on tannins) is provided. In vitro studies show that the solubility of salivary protein-tannin complexes in gastric conditions depends on the degree of tannin polymerization, while complexes are partly solubilized by bile salts. Salivary proteins-polyphenols interactions may affect digestive processes. For example, polyphenols can bind to and inhibit salivary amylase, with downstream consequences on starch digestion. Some salivary proteins (PRPs) prevent tannin-induced reduced protein digestibility, probably through binding tannins before they interact with digestive proteases. Salivary proteins may also act as scavenger molecules to limit the intestinal uptake of tannins.

NMR structural elucidation of dehydrodimers resulting from oxidation of 5-O-caffeoylquinic acid in an apple juice model solution.

During apple juice and cider-making processes, phenolic compounds undergo enzymatic oxidation. 5-O-caffeoylquinic acid (CQA) is one of the major hydroxycinnamic acid derivatives and it is the preferential substrate for polyphenol oxidase (PPO) in apple juices. Consequently, CQA dehydrodimers (MW 706 Da) are among the main products resulting from CQA oxidation. CQA dehydrodimers were previously synthesized in a biomimetic apple juice model solution. Following their purification and characterization using UV-Visible spectra and mass spectrometry, the structures of seven CQA dehydrodimers were elucidated using 1H and 13C one- and two-dimensional NMR spectroscopy. Six of them exhibited dihydrobenzofuran, benzodioxane, or dihydronaphtalene skeletons, which are caffeicin-like structures. Interestingly, a new dehydrodicaffeoyldiquinic acid molecule was also characterised for which two novel structures showing a symmetric dicatechol skeleton were also proposed.

Biochemical and physical-chemical characterisation of leaf proteins extracted from Cichorium endivia leaves.

This study provides a detailed characterisation of a leaf protein concentrate (LPC) extracted from Cichorium endivia leaves using a pilot scale process. This concentrate contains 74.1% protein and is mainly composed of Ribulose-1,5-BISphosphate Carboxylase/Oxygenase (RuBisCO). We show that the experimentally determined extinction coefficient (around 5.0 cm-1 g-1 L depending on the pH) and refractive index increment (between 0.27 and 0.39 mL g-1) are higher than the predicted ones (about 1.6 cm-1 g-1 L and 0.19 mL g-1, respectively). In addition, the UV-visible absorption spectra show a maximum at 258 nm. These data suggest the presence of non-protein UV-absorbing species. Chromatographic separation of the concentrate components in denaturing conditions suggests that RuBisCO SC may be covalently bounded to few phenolic compounds. Besides, the solubility of LPC proteins is higher than 90% above pH 6. Such high solubility could make LPC a good candidate as a functional food ingredient.

Preparative isolation of apple flavan-3-ols monomers and oligomers using pH-zone-refining centrifugal partition chromatography combined with reversed-phase liquid chromatography.

Flavan-3-ols (catechin monomers and procyanidins) are the main class of polyphenols in apples and are found in high concentrations in cider apple varieties. They are known to be involved in bitterness and astringency in apple-based beverages, and also contribute to polyphenol nutritional intake.Therefore, highly purified flavan-3-ol fractions isolated from raw materials are needed to study their various properties. For this purpose, a gentle strategy combining pH-zone-refining centrifugal partition chromatography (pH-ZRCPC) and preparative reversed-phase liquid chromatography (Prep-RPLC) was developed to recover one hundred milligrams of a high purity apple flavan-3-ol fraction. First, pH-ZRCPC fractionation in descending mode was optimized to remove hydroxycinnamic acid derivatives using a biphasic mixture composed of ethyl acetate/n-butanol/water (3/2/5, v/v). Trifluoroacetic acid and sodium hydroxide were used as retainer and eluter, in the upper and lower phases, respectively. Secondly, Prep-RPLC separation was carried out in isocratic mode at 20% ACN to remove dihydrochalcones. Finally, from one gram of a crude polyphenol extract, four hundred and nine milligrams of a highly purified fraction of flavan-3-ols with an average degree of polymerization close to 3.1 was obtained with 73% recovery.

Interactions between polyphenol oxidation products and salivary proteins: Specific affinity of CQA dehydrodimers with cystatins and P-B peptide.

Throughout the apple juice and cider making process, polyphenols undergo enzymatic oxidation which generates a great variety of polyphenol oxidation products. Since 5'-O-Caffeoylquinic acid (CQA) is one of the major phenolic compounds and the preferential substrate for polyphenoloxidase in apple juice, its oxidation leads to the formation of newly formed molecules by which dehydrodimers (MW 706 Da) are included. Interactions of salivary proteins (SP) with native polyphenols is a well-known phenomenon, but their interactions with polyphenol oxidation products has not been studied yet. In this work, we decided to decipher the interactions between CQA dehydrodimers and SP (gPRPs, aPRPs, statherins/P-B peptide, and cystatins) using HPLC-UV and fluorescence. These results showed that contrary to what was expected, CQA dehydrodimers presented a low interaction with PRPs, but revealed a specific interaction with statherins/P-B peptide and cystatins. This work settles for the first time the interactions between SP and polyphenol oxidation products.

Structural characterization of native and oxidized procyanidins (condensed tannins) from coffee pulp (Coffea arabica) using phloroglucinolysis and thioglycolysis-HPLC-ESI-MS.

Procyanidins from coffee pulp are responsible from the limited valorization of this by-product. Information about procyanidin structure is still scarce and imprecise. The aim of this work was to study the native and oxidized procyanidins from coffee pulp with respect to composition and structure. An aqueous acetone extract from coffee pulp was purified using Sephadex LH-20. Butanolysis, phloroglucinolysis and thioglycolysis coupled to HLPC-ESI-MS were applied for the characterization of the native and oxidized procyanidins. The purification allowed to recovery three fractions (aqueous, ethanolic and acetonic) and only acetone fraction showed a high concentration of procyanidins (98%, w/w). HPLC-ESI-MS of procyanidins-rich fraction without any reaction resulted in a UV-Vis chromatogram unresolved typical of the presence of procyanidins. The extracted ion chromatogram and MS2 analysis revealed the presence from dimers to pentamers of native procyanidins. Interestingly, by first time an A-type trimeric procyanidin (m/z of 863) was observed in coffee pulp. In our study, (-)-epicatechin was the constitutive unit of procyanidins with an aDP of 6.8 (oligomeric native procyanidins) according to the phloroglucinolysis assay. Two oxidation markers useful to characterization of oxidized procyanidins were observed in the procyanidins-rich fraction after thioglycolysis, a dimer A2-ext and a molecule that corresponds to a linkage between an extension and a terminal unit. Coffee pulp procyanidins were presented with only a minor class of oxidized procyanidins. As far as we know, this is the first study about characterization of the oxidized procyanidins from coffee pulp.

Data set on hydroxycinnamic acid ester analysis from the cell walls of apples and grapes.

Data on the esters of hydroxycinnamic acids (HCAs) from the cell walls of wine grapes (Cabernet franc) and cider apples (Douce Moen and Guillevic) were acquired. Caffeic acid, p-coumaric acid (pCA) and ferulic acid (FA) monomers were identified by HPLC-UV/MS. Means to limit the oxidative degradation during cell wall preparation were assessed by the yield of HCA recovered after alkaline extraction. Following the optimum cell wall preparation, the pCA content varied between 2.3 and 32.5 mg kg-1 dry cell wall and that of FA varied between 0.3 and 17.2 mg kg-1 dry cell wall. Higher HCA quantities were found in the peels compared to the flesh and in apples compared to grapes. The Douce Moen apple was richer in HCAs than the Guillevic apple. pCA was localized in the cell wall as observed by TEM after labeling with the INRA-COU1 antibody that recognizes pCA linked to O-5 of arabinose. The anti-FerAra antibody targeting FA on O-5 of arabinose failed to locate FA esters in the apple and grape cell walls.

Heat-unstable apple pathogenesis-related proteins alone or interacting with polyphenols contribute to haze formation in clear apple juice.

Physico-chemical instability is a damaging defect that can occur in clear bottled beverages leading to the formation of haze. In a previous study, we showed the presence of proteins in haze gathered from apple juices. For the first time, proteomics was used to sequence and identify four pathogenesis-related proteins (PRPs) from the haze of a commercial apple juice. Then, a study involving purified PRPs and polyphenols from apple juice was conducted in model solution to understand the mechanisms by which they are involved in haze formation. Visual assessment revealed that apple juice pathogenesis-related proteins are able to form haze alone when thermally denatured. These proteins were also able to interact with apple juice procyanidins to form complexes that can be precipitated using ultracentrifugation, even without prior heating. These interactions were greater when the degree of polymerization of tannins increased.

Rigidity, conformation, and solvation of native and oxidized tannin macromolecules in water-ethanol solution.

We studied by light scattering and small angle x-rays scattering (SAXS) conformations and solvation of plant tannins (oligomers and polymers) in mixed water-ethanol solutions. Their structures are not simple linear chains but contain about 6% of branching. Ab initio reconstruction reveals that monomers within a branch are closely bound pairwise. The chains are rather rigid, with the Kuhn length b = 13+/-3 nm, corresponding to about 35 linearly bound monomers. Contribution of solvation layer to SAXS intensity varies in a nonmonotonic way with ethanol content phi(A), which is an indication of amphipathic nature of tannin molecules. Best solvent composition phi(A)(B) is a decreasing function of polymerization degree N, in agreement with increasing water solubility of tannins with N. Polymers longer than b present a power-law behavior I approximately Q(-d) in the SAXS profile at high momentum transfer Q. The monotonic decrease in d with increasing phi(A) (from 2.4 in water to 1.9 in ethanol) points that the tannins are more compact in water than in ethanol, presumably due to attractive intramolecular interactions in water. Tannins were then oxidized in controlled conditions similar to real biological or food systems. Oxidation does not produce any intermolecular condensation, but generates additional intramolecular links. Some oxidation products are insoluble in water rich solvent. For that reason, we identify these species as a fraction of natural tannins called "T1" in the notation of Zanchi et al. [Langmuir 23, 9949 (2007)]. Within the fraction left soluble after oxidation, conformations of polymeric tannins, despite their higher rigidity, remain sensitive to solvent composition.