Celestino Santos-Buelga

Proanthocyanidins and tannin-like compounds - nature, occurrence, dietary intake and effects on nutrition and health

Proanthocyanidins (syn condensed tannins) are complex flavonoid polymers naturally present in cereals, legume seeds and particularly abundant in some fruits and fruit juices. They share some common structural features—phenolic nature and high molecular weight—with phenolic polymers found in black tea and red wine (called here tannin-like compounds). The polymeric nature of proanthocyanidins makes their analysis and estimation in food difficult. For this reason, little is known about their consumption, although they likely contribute a large part of the daily polyphenol intake. They also share common physicochemical properties: they form stable complexes with metal ions and with proteins and are, like other polyphenols, good reducing agents. Many of their biological effects of nutritional interest derive from these properties. As metal ion chelators, they influence the bioavailability of several minerals. The nutritional significance of the non-specific complexation of proteins is less clear. As reducing agents, they may participate in the prevention of cancers, both of the digestive tract and inner organs. They may also protect LDLs against oxidation and inhibit platelet aggregation and therefore prevent cardiovascular diseases. In vitro, animal and human studies on the prevention of these chronic diseases are reviewed with particular attention to wine and tea polyphenols. The lack of data on their bioavailability and the paucity of human studies are emphasised. © 2000 Society of Chemical Industry

High-Flavonol Tomatoes Resulting from the Heterologous Expression of the Maize Transcription Factor Genes LC and C1

Flavonoids are a group of polyphenolic plant secondary metabolites important for plant biology and human nutrition. In particular flavonols are potent antioxidants, and their dietary intake is correlated with a reduced risk of cardiovascular diseases. Tomato fruit contain only in their peel small amounts of flavonoids, mainly naringenin chalcone and the flavonol rutin, a quercetin glycoside. To increase flavonoid levels in tomato, we expressed the maize transcription factor genes LC and C1 in the fruit of genetically modified tomato plants. Expression of both genes was required and sufficient to upregulate the flavonoid pathway in tomato fruit flesh, a tissue that normally does not produce any flavonoids. These fruit accumulated high levels of the flavonol kaempferol and, to a lesser extent, the flavanone naringenin in their flesh. All flavonoids detected were present as glycosides. Anthocyanins, previously reported to accumulate upon LC expression in several plant species, were present in LC/C1 tomato leaves but could not be detected in ripe LC/C1 fruit. RNA expression analysis of ripening fruit revealed that, with the exception of chalcone isomerase, all of the structural genes required for the production of kaempferol-type flavonols and pelargonidin-type anthocyanins were induced strongly by the LC/C1 transcription factors. Expression of the genes encoding flavanone-3′-hydroxylase and flavanone-3′5′-hydroxylase, which are required for the modification of B-ring hydroxylation patterns, was not affected by LC/C1. Comparison of flavonoid profiles and gene expression data between tomato leaves and fruit indicates that the absence of anthocyanins in LC/C1 fruit is attributable primarily to an insufficient expression of the gene encoding flavanone-3′5′-hydroxylase, in combination with a strong preference of the tomato dihydroflavonol reductase enzyme to use the flavanone-3′5′-hydroxylase reaction product dihydromyricetin as a substrate.

Evaluation of the antioxidant properties of fruits

Abstract Twenty-eight different fruits were analysed for antioxidant activities using two different methods, one that determines the inhibition of ascorbate/iron-induced peroxidation of phosphatidylcholine, by means of thiobarbituric acid-reactive substances (TBARS) measurement, and another that evaluates the scavenging of the radical cation of 2,2′-azinobis (3-ethyl-benzothiazoline-6-sulphonate) (ABTS) relative to Trolox C, a water-soluble vitamin E analogue. The values of the antioxidant capacities of the analysed fruits were in a wide range from 50 g to more than 5 mg for the IW 50 (dry weight causing 50% inhibition of lipidic peroxidation) and from it 406 μmol/g for the TEAC (μmol Trolox equivalents g −1 ). There was no significant correlation between antioxidant activity (IW 50 and TEAC) and the contents of flavanol of the samples. The antioxidant activities of the analysed fruits cannot only be attributed to their flavanol contents, but to the result of the action of different antioxidant compounds present in the fruits and to possible synergic and antagonist effects still unknown.

Antihypertensive effects of the flavonoid quercetin

The blood pressure lowering effect of a fruit and vegetable-rich diet is a necessary dietary lifestyle measure now included the guidelines for the management of arterial hypertension. Furthermore, flavonoids represent a major class of plant polyphenolics. The present review addresses the antihypertensive effect of quercetin, one of the most abundant flavonoids present in fruits and vegetables, and probably the best studied flavonoid because of its high biological activity. Quercetin has been shown to induce a progressive, dose-dependent and sustained reduction in blood pressure when given chronically in several rat models of hypertension, including spontaneously hypertensive rats, L-NAME-treated rats, DOCA-salt hypertensive rats, two-kidney one-clip Goldblatt rats, rats with aortic constriction and Dahl salt-sensitive hypertensive rats. Quercetin was also effective in reducing blood pressure in rat models of metabolic syndrome, including the obese Zucker rats as well as rats treated with a high-sucrose, high-fat diet. Quercetin also prevented morphological and functional changes in the heart, vessels and kidney, while increasing production of reactive oxygen species associated with hypertension. A high dose of quercetin also reduced blood pressure in stage 1 hypertensive patients in a randomized, double-blind, placebo-controlled, crossover study. Since raised blood pressure is the major cause of stroke as well as an important risk factor for ischemic heart disease, we propose that the blood pressure-lowering effect of quercetin could be an important mechanism contributing to the reduced risk of myocardial infarction and stroke observed with fruit and vegetables-rich diets, and possibly with flavonoid-rich diets.

One-month strawberry-rich anthocyanin supplementation ameliorates cardiovascular risk, oxidative stress markers and platelet activation in humans

Strawberries are an important fruit in the Mediterranean diet because of their high content of essential nutrients and beneficial phytochemicals, which seem to exert beneficial effects in human health. Healthy volunteers were supplemented daily with 500 g of strawberries for 1 month. Plasma lipid profile, circulating and cellular markers of antioxidant status, oxidative stress and platelet function were evaluated at baseline, after 30 days of strawberry consumption and 15 days after the end of the study. A high concentration of vitamin C and anthocyanins was found in the fruits. Strawberry consumption beneficially influenced the lipid profile by significantly reducing total cholesterol, low-density lipoprotein cholesterol and triglycerides levels (-8.78%, -13.72% and -20.80%, respectively; P<.05) compared with baseline period, while high-density lipoprotein cholesterol remained unchanged. Strawberry supplementation also significant decreased serum malondialdehyde, urinary 8-OHdG and isoprostanes levels (-31.40%, -29.67%, -27.90%, respectively; P<.05). All the parameters returned to baseline values after the washout period. A significant increase in plasma total antioxidant capacity measured by both ferric reducing ability of plasma and oxygen radical absorbance capacity assays and vitamin C levels (+24.97%, +41.18%, +41.36%, respectively; P<.05) was observed after strawberry consumption. Moreover, the spontaneous and oxidative hemolysis were significant reduced (-31.7% and -39.03%, respectively; P<.05), compared to the baseline point, which remained stable after the washout period. Finally, strawberry intake significant decrease (P<.05) the number of activated platelets, compared to both baseline and washout values. Strawberries consumption improves plasma lipids profile, biomarkers of antioxidant status, antihemolytic defenses and platelet function in healthy subjects, encouraging further evaluation on a population with higher cardiovascular disease risk.

Chromatic characterization of anthocyanins from red grapes-I. pH effect

Abstract Five anthocyanins (the 3-monoglucoside of delphinidin, cyanidin, petunidin, peonidin and malvidin), isolated from red grape skins, were subjected to a spectroscopic study to characterize their chromatic properties in a model solution imitating wine in the pH range 1.5–7.0. Tristimulus colorimetry (CIEXYZ, CIELUV and CIELAB colour spaces) proved to be a useful tool for the chromatic characterization of these anthocyanins. Significant colour differences were found among them, related to the number and type of substituents in their B rings. Two-substituted compounds were located in the area of orange hue, while three-substituted compounds were in the area of red–purple. As the methoxylation degree increased, a shift was observed toward purple. Also, a relationship was found between saturation and chroma and the number of hydroxy groups in the B ring. In all the anthocyanins studied, the increase in pH provoked a curve displacement in chromaticity, approaching that of the illuminant..

Phenolic acids determination by HPLC-DAD-ESI/MS in sixteen different portuguese wild mushrooms species

Analysis of phenolic compounds in sixteen Portuguese wild mushrooms species has been carried out by high-performance liquid chromatography coupled to photodiode array detector and mass spectrometer (HPLC-DAD-ESI/MS). No flavonoids were detected in the analysed samples, but diverse phenolic acids namely protocatechuic, p-hydroxybenzoic and p-coumaric acids, and two vanillic acid isomers were found and quantified. A related non-phenolic compound, cinnamic acid, was also detected in some samples, being the only compound found in Cantharellus cibarius (14.97 mg/kg, dry matter), Lycoperdon perlatum (14.36 mg/kg) and Macrolepiota procera (21.53 mg/kg). p-Hydroxybenzoic acid was found in the majority of the samples, being the most abundant compound in Agaricus silvicola (238.7 mg/kg). Ramaria botrytis showed the highest phenolic acids concentration (356.7 mg/kg) due to the significant contribution of protocatechuic acid (342.7 mg/kg).

Developmental changes in the phenol concentrations of ‘Golden delicious’ apple fruits and leaves

The phenolic content of apple fruit skin and leaves was determined at the developmental stage of each organ. Phenolic levels decreased on a dry weight basis during the seasonal development of fruits and leaves with respect to their ontogenesis but the single compounds did not behave uniformly. A shift in flavanol pools from monomeric to oligomeric structures during fruit growth indicated the biosynthetic tendency towards the formation of procyanidins at the end of the growing period. Among the procyanidins identified epicatechin-(4 beta,6)-epicatechin-(4 beta,6)-epicatechin has not been reported previously from apple.

Recent advances in polyphenol research

This important book covers polyphenol chemistry, biosynthesis and genetic manipulation, ecology and plant physiology, food and nutritional aspects and the effects of polyphenols on health. Included within the contents are cutting edge chapters on biotic and abiotic stress in plants, safety and toxicity in foods, functionality and nutraceutical benefits in nutrition, and aspects of pharmaceutical and cosmetic discovery and development.

Comparative flavan-3-ol composition of seeds from different grape varieties

The flavanol composition of the seeds of 17 varieties of grape cultivated in the main wine-producing areas of Castilla-Leon (Spain) was studied. Twenty-seven different flavan-3-ols of procyanidin type were found, but no prodelphinidins were detected. Minor amounts of four hydrolyzable tannins were also present in the seed extracts of most of the grapes analyzed. All the varieties contained galloyled flavanols, such that their presence could be considered characteristic of the grape seed composition. Some proposals concerning the biosynthesis of flavan-3-ol in the grape seed are also advanced taking into account the compounds found.