Wine Constituents Research Articles

Effects of red wine, alcohol, and quercetin on coronary resistance and conductance arteries.

Moderate consumption of alcoholic beverages is associated with a reduced risk of coronary heart disease (CHD). Some evidence suggests that red wine is particularly beneficial in this regard and may account in part for the French paradox, although the mechanism of this effect is unknown. We assessed the effects of red wine, ethanol, and quercetin, a major flavonoid constituent of red wine, in coronary resistance vessels (80-150 microm, i.d.) and conductance vessels (300-525 microm, i.d.) of the rabbit. Vessel wall tension was measured in isolated segments maintained in a wire-type myograph (37 degrees C) and preconstricted with 30 mM K+. At an alcohol concentration (14 mM) equivalent to moderate consumption, red wine evoked a small, transient constrictor effect in resistance and conductance vessels (9+/-4%, n = 5; 8+/-1%, n = 7, respectively; p < 0.05). Ethanol alone at this concentration was without effect. Quercetin (5.6, 8, and 30 microM) significantly relaxed resistance (-32+/-4%, n = 10; -47+/-2%, n = 7; -82+/-6%, n = 8, respectively) and conductance (-20+/-3%, n = 8; -32+/-4%, n = 8; -72+/-7%, n = 8, respectively) coronary arteries. Vasorelaxation by quercetin was endothelium-independent and was significantly greater in resistance than in conductance vessels. These data suggest that red wine and ethanol do not evoke relaxation in small coronary arteries at concentrations associated with moderate consumption. Quercetin elicits marked coronary vasorelaxation that is endothelium-independent. However, the concentrations of quercetin necessary to achieve this action are not attained with moderate red wine consumption.

Analysis of phenolic constituents of biological interest in red wines by high-performance liquid chromatography

We describe a reversed-phase HPLC method that uses gradient elution and diode array detection to determine four biologically active phenolic constituents of red wines: gallic acid, trans-resveratrol, quercetin and rutin. The method permits direct injection without sample pre-treatment. ODS Hypersil served as the stationary phase; the gradient was formed by acetic acid, methanol, and water. Each analysis required an equilibration period of 10 min and a run time of 50 min for completion. Previously, total phenols were analysed according to the Folin–Ciocalteu method, using gallic acid as the standard, and the results are given as gallic acid equivalent.

Isolation and characterization of novel benzoates, cinnamates, flavonoids, and lignans from Riesling wine and screening for antioxidant activity.

A German Riesling wine has been fractionated with the aid of countercurrent chromatography. After purification by HPLC, the structures of 101 compounds were established by mass spectrometry and NMR spectroscopy. Seventy-three of the isolated compounds exhibited a phenolic or benzylic structure. Fifty-four compounds were reported for the first time as Riesling wine constituents. New compounds identified in this work included twelve benzoic and cinnamic acid derivatives. In addition to two isomeric (E)-caffeoyl ethyl tartrates, the glucose esters of (E)-cinnamic, (E)-p-coumaric, and (E)-ferulic acid, as well as the 4-O-glucosides of (E)- and (Z)-ferulic acid, have been identified for the first time in Riesling wine. The structures of two additional phenylpropanoids were elucidated as 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-propan-1-one and 2,3-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-propan-1-one. Moreover, two ethyl esters, i.e., ethyl protocatechuate and ethyl gallate, as well as the glucose ester of vanillic acid, were newly detected in Riesling wine. Novel representatives in the flavonoid group were dihydrokaempferol, dihydroquercetin, and four dihydroflavonol glycoconjugates, i.e., the 3-O-glucosides of dihydrokaempferol and dihydroquercetin, as well as the 3-O-xyloside and the 3'-O-glucoside of dihydroquercetin. Additionally, six novel lignans, i.e., lariciresinol 4-O-glucoside, three isolariciresinol derivatives, and two secoisolariciresinols, as well as three neolignans were isolated. Structural elucidation of the newly isolated wine constituents is reported together with the determination of their antioxidant activity.

Membrane fouling by turbidity constituents of beer and wine: characterization and prevention by means of infrasonic pulsing

During microfiltration of fermented beverages such as beer and wine, colloidal turbidity constituents play an important role in membrane fouling and therefore hinder the permeate flux. This paper studies polymeric membrane fouling by turbidity constituents of beer and wine and proposes a fouling prevention method to improve permeate flux. The model turbidity suspensions, based on gelatin and tannic acid, were microfiltered in a stirred cell using 0.2 μm cellulose acetate (CA), polyvinylidene di-fluoride (PVDF) and polycarbonate (PC) membranes under constant pressure (0.79 bar) and agitation (1350 rpm). Fouling caused by either a separate protein solution or a separate polyphenol solution was considerably lower than the fouling caused by a solution of both at the same concentration, which suggests that fouling is mainly caused by light scattering complexes formed by polyphenols and proteins. In all cases, during the microfiltration of model turbidity suspensions, the internal fouling mechanism dominated first and this was later followed by a period of external fouling. It was shown that membrane fouling depends strongly on the initial turbidity of the solution. A lower turbidity of the model solution resulted in a lower final total resistance. The initial turbidity also influenced the duration of the initial period of internal fouling. For the lower turbidity model solutions internal fouling lasted longer. An infrasonic pulsing was applied to remove foulant cake and improve permeate flux during microfiltration of model turbidity suspensions, and wine and beer samples. In infrasonic pulsing, high frequency pulses of permeate are periodically sent back in the direction of the membrane at a lower pressure than the transmembrane pressure. This causes the membrane to vibrate rapidly and removes a portion of the foulant cake. The model turbidity suspension (33±2 NTU) was filtered through a flat-sheet PVDF membrane at a transmembrane pressure of 0.65 bar. In the experiments with infrasonic pulsing a 0.2 duty cycle ( Φ) was used. The improvement in net permeate flux depended on the infrasonic pulse frequency. For the highest frequency (6.67 Hz) net permeate flux improved fourfold in comparison with long-term permeate flux in normal cross-flow. These data fitted very well to the theoretical model first published by Czekaj [Reducción del Ensuciamiento de Membranas Poliméricas en Procesos de Microfiltración Tangencial de Suspensiones Biológicas Mediante Utilización de Infrasonidos y de la Técnica de Flujo Transversal, PhD Thesis, Universitat Rovira i Virgili, Spain] Experiments with wine and beer samples filtered through a PS hollow fiber membrane yielded approximately 2.4 and 2.1 times larger permeate fluxes with infrasonic pulsing than with normal cross-flow.

Centrifugal precipitation chromatography -- a novel chromatographic system for fractionation of polymeric pigments from black tea and red wine.

A novel chromatographic system was developed and first applied to the fractionation of polymeric pigments from black tea and red wine. Centrifugal precipitation chromatography (CPC) generates solvent gradients through a long separation channel under a centrifugal force field. Tea and wine extracts are precipitated in a hexane- or methyl tert-butyl ether-rich environment and are exposed to a gradually increasing ethanol concentration. This causes a repetitive precipitation and dissolution of the biopolymers along the channel. Consequently, they are eluted in the order of their solubility in the organic solvent. It is shown by HPLC analysis of the separated fractions that monomers elute first, whereas fractionated polymers can be found at the end of the chromatographic run. This novel method allows gentle fractionation of polymeric tea and wine constituents and also has potential for use in preparative-scale separations.

Differential response of immature rat uterine tissue to ethinylestradiol and the red wine constituent resveratrol.

The stilbene derivative resveratrol (RES) is a phytoalexin of grapes, peanuts and other fruits. It is structurally related to stilbene estrogens and an estrogenic potential of RES has recently been demonstrated in a number of in vitro studies. In this investigation, the uterotrophic responses of immature Wistar rats to subcutaneous administration of RES (18, 58, and 575 mg/ kg) and the reference estrogen ethinylestradiol (EE2; 0.3, 1, 3, 30 microg/kg) on three consecutive days were determined. Uterine weight, histopathological changes, immunohistochemical expression of nuclear estrogen receptor-alpha (ERalpha) and progesterone receptor (PR) protein, gene expression of ERalpha and PR at the messenger ribonucleic acid (mRNA) level and peroxidase induction were examined. EE2 dose dependently increased uterine weight, enlarged the uterine lumen and induced hypertrophy of epithelial, stromal and myometrial cells. Expression of ERalpha protein in epithelial, stromal and myometrial nuclei and of PR protein in epithelial nuclei was reduced in EE2-treated rats, while PR protein in stromal and myometrial nuclei was increased in a dose-dependent manner. EE2 increased messenger ribonucleic acid (mRNA) levels of uterine PR and induced peroxidase activity. In contrast, RES rather mildly decreased uterine weight, while histology did not reveal differences between controls and RES-treated rats. Expression of nuclear ERalpha protein was dose dependently decreased in epithelial, stromal and myometrial cells of RES-treated rats, while nuclear PR protein content was similar in controls and RES-treated rats. Following administration of RES, a trend toward reduced levels of ERalpha and PR mRNA was found, while no peroxidase induction occurred. Plasma levels of RES, 45 min after the administration of a single subcutaneous dose of 500 mg/kg, were in the range 1-2 microM. In summary, an estrogenic potential of RES could not be substantiated in this in vivo study, although the most effective route of administration and extremely high doses were used and plasma levels were in the range reported to be effective in vitro. Whether other pharmacological properties of RES could mediate the observed changes in RES-treated animals is discussed.

The world of resveratrol.

Since the discovery of trans-resveratrol (3,5,4’-trihydroxystilbene) as a constituent of wine by Siemann and Creasy, first reported in 1992 (1), the possibility that this compound, almost unique to red wine among constituents of the human diet, may in large measure account for the putative health benefits of this beverage beyond its mere content of vulgar ethanol, excited the imagination of the scientific and medical communities, initiating a ferment of research and enquiry that continues to this day. Indeed, ripples of these activities from time to time flow into the pages of the lay press, so that resveratrol has become a molecule impacting the consciousness of many well-informed members of the lay public. In March, 1997 we published a major review incorporating 183 references forming the bulk of the world literature on resveratrol up to that time (2). Our bottom line was that the future of resveratrol did not look particularly promising given the reality that, despite its miraculous performances in the culture dish and the test tube, the intact bodies of mice and men proved to be an inhospitable millieu robbing it of its presumed powers. To us, it seemed to be a compound for whom the bell tolled , but others did not share this gloomy prognosis. In fact, so much new work on this topic has been published in a mere two years-and-a-bit that a reappraisal of the situation deserves a welcome and is mandated by the present Symposium. Resveratrol exists as trans and cis isomers. Very little is known about the latter. When the nature of resveratrol is not specified, the reader should assume that the text refers to the trans isomer.

Examination of non-volatile organic compounds in red wines made in Eger

At present the analytical investigation of human consumption products, with special regard to the environmental and health connections, is basically important all over the world. Recently in several countries it was almost impossible to sell wine without a certificate of quality, based on modern, instrumental analytical methods. There is well-known medical–biological evidence, which has proved the antioxidant and vein wall-protecting effects of the wines in the case of frugal ingestion. On the other hand, they also play a part in the prevention of heart attacks. Regarding these biological effects the most important constituents of wines are the flavonoids, anthocyanidins, and their glycosides. Anthocyanins can be identified first of all in red wines. The organic constituents have characteristic antioxidant effects, which can play an important role in health protection. During our investigation we have studied the volatile and non-volatile organic compounds in different types of wines made in Eger and Tokay (Hungary). In our opinion these types of research projects have unique importance, from the economic viewpoint and in that they are not negligible in a national context. The separation and determination of volatile compounds was carried out by applying a Finnigan GSQ GC-MS apparatus and the non-volatile ones with HPLC-DAD and FAB MS techniques.

Resveratrol, a red wine constituent, is a mechanism-based inactivator of cytochrome P450 3A4

Resveratrol, a phytoalexin found in red wine, has been shown to possess antioxidant and antimutagenic properties. Incubation of resveratrol with Sf 9 insect microsomes containing baculovirus-derived human cytochrome P450 3A4 (CYP3A4) and NADPH-cytochrome P450 reductase showed that resveratrol inactivated CYP3A4 in a time- and NADPH-dependent manner. Resveratrol, erythromycin and troleandomycin inactivated CYP3A4 at a similar rate (as reflected by k inact) whereas the binding affinity to CYP3A4 (as reflected by K I) was in the order of: troleandomycin > erythromycin > resveratrol. (K I and k inact for CYP3A4 inactivation by resveratrol, erythromycin and troleandomycin are 20 μM and 0.20 min −1, 5.3 μM and 0.12 min −1 and 0.18 μM and 0.15 min −1, respectively.) Fractionation studies of red wine showed that fractions that did not contain resveratrol inactivated CYP3A4 significantly. In addition, the resveratrol content in red wine used in the study was too low to account for the degree of CYP3A4 inactivation observed after red wine treatment. Inactivation studies using a variety of red wine types showed that the CYP3A4 inactivation did not correlate to their resveratrol content. In summary, data here showed that resveratrol is an effective mechanism-based inactivator of CYP3A4; however, it is not one of the main red wine constituents that are responsible for CYP3A4 inactivation by red wine. Nevertheless, inactivation of CYP3A4 by resveratrol may cause clinically relevant drug interactions with CYP3A4 substrates.

Neuroprotective abilities of resveratrol and other red wine constituents against nitric oxide-related toxicity in cultured hippocampal neurons.

Animal and epidemiological studies suggest that polyphenol constituents of red wine possess antioxidant activities that favour protection against cardiovascular disease - the so-called. 'French paradox' - and possibly, central nervous system disorders such as Alzheimer's disease (AD) and ischaemia. In the present study, the potential of three major red wine derived-polyphenols to protect against toxicity induced by the nitric oxide free radical donors sodium nitroprusside (SNP) and 3-morpholinosydnonimine (SIN-1) was examined in cultured rat hippocampal cells. Both co- and post-treatments with either the stilbene resveratrol (5 - 25 microM) or the flavonoids quercetin (5 - 25 microM) and (+)-catechin (1 - 10 microM) were capable of attenuating hippocampal cell death and intracellular reactive oxygen species accumulation produced by SNP (100 microM and 1 mM, respectively). However, among the phenolic compounds tested, only the flavonoids afforded significant protection against 5 mM SIN-1-induced toxicity. The effects of phenolic constituents were shared by Trolox (100 microM), a vitamin E analogue, but not by selective inhibitors of cyclo-oxygenases (COX) and lipoxygenases (LOX). Among the phenolic compounds tested, only quercetin (10 microM) inhibited 100 microM SNP-stimulated protein kinase C (PKC) activation, whereas none of them were able to attenuate nitrite accumulation caused by SNP (100 microM). Taken together, these data suggest that the neuroprotective abilities of quercetin, resveratrol, and (+)-catechin result from their antioxidant properties rather than their purported inhibitory effects on intracellular enzymes such as COX, LOX, or nitric oxide synthase. Quercetin, however, may also act via PKC to produce its protective effects.

Resveratrol, a red wine constituent, has anticarcinogenic properties

Resveratrol, a red wine constituent, has anticarcinogenic properties

Synthesis of Enantiomerically Pure (−)-Wine Lactone Based on a Palladium-Catalyzed Enantioselective Allylic Substitution

The first enantioselective synthesis of enantiomerically pure (−)-wine lactone, (−)-1a, a fragrance constituent of various white wines, and its epimer (+)-1b, was carried out. The key steps are allylic substitution of (±)-2-cyclohexen-1-yl acetate (2) with dimethylmalonate using palladium complexes of phosphanyldihydrooxazol L1 or of the phosphanylcarboxylic acid L2 as catalyst, subsequent decarboxylation, iodolactonization and elimination, furnishing enantiomerically pure bicyclic lactone (+)-7 in 47% overall yield. The diastereoselective introduction of methyl groups by SN2′-type substitution with an organocopper compound and by enolate alkylation gave lactone (−)-1a in 43% overall yield from (+)-7.

Assay of Resveratrol and Derivative Stilbenes in Wines by Direct Injection High Performance Liquid Chromatography

The concentrations of <i>trans</i>-and <i>cis</i>-isomers of four stilbenes produced by grapevines — piceid, resveratrol, resveratrol dehydrodimer, and pterostilbene — were determined in wines from Burgundy and Switzerland. Analyses were carried out by direct injection high performance liquid chromatography. All red wines studied contained high levels of piceid and resveratrol, but no pterostilbene. Resveratrol dehydrodimer was also detected. It is the first time that this compound is reported as a constituent of wine.

Identification of some volatile descriptors of the rock-rose-like aroma of fortified red wines from douro demarcated region.

2,2,6-Trimethylcyclohexanone and ethyl dihydrocinnamate, known sensorial descriptors of rock-rose (Cistus ladaniferus) aroma, are reported as minor constituents of young fortified wines from the Douro Demarcated region (vintage 1997) for the first time. These compounds were respectively identified by GC-MS in two different fractions (A and B) obtained by preparative GC. Quantitative analysis by GC-MS was performed on 14 fortified wines originating from the different Douro subregions. Ethyl dihydrocinnamate was found to be present at concentrations between 2.3 and 6.7 microgram/L, while 2,2,6-trimethylcyclohexanone ranged from 0.05 to 0.4 microgram/L.

Effect of antioxidants on the flavor characteristics and the gas chromatography/olfactometry profiles of champagne extracts.

This paper aims to investigate the possibility of oxidation of the flavor active constituents of champagne wines during the critical steps of the extraction for analytical purposes and to estimate its consequences on the extract's aroma characteristics. The investigation uses conventional sensory and GC/O analyses. Triangle and similarity sensory tests carried out on champagne samples and on their extracts show that the absence of antioxidants implies a deterioration of the aroma quality. Conversely, the use of 2-tert-butyl-4-methoxyphenol (BHA) as antioxidant gives an extract with an odor judged to be more similar to the original odor of the champagne. The alteration in the odor of the unprotected extracts is explained, from GC/O and GC/MS analyses, as three types of uncontrolled reactions. These are, first, the chemical oxidation of the fusel alcohols and the amino acids; second, the oxidation of sensitive flavor compounds such as enolic lactones and monoterpenic alcohols; and, third, the hydrolysis of fatty acid esters and saturated gamma-lactones. A systematic use of BHA during flavor extraction is therefore advisable, in particular when oxidation phenomena such as wine aging are under study.

Liquid chromatography/mass spectrometric determination of trans-resveratrol in wine using a tandem solid-phase extraction method.

Liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization has been successfully applied to the determination of trans-resveratrol (3,5,4'-trihydroxystilbene) in wine. Of a range of analytical conditions that were tested, optimum results were obtained by the use of reversed-phase high performance liquid chromatography (HPLC) using a mixture of methanol and ammonium acetate as the mobile phase. The negative-ion spectrum of trans-resveratrol showed pseudo-molecular ion, [M - H](-), which was the most abundant ion, and low fragment ions corresponding to the losses of hydroxyl groups of the phenol nucleus. Enhanced selectivity for the separation between trans-resveratrol and endogenous wine constituents was afforded by sample purification with a tandem solid-phase extraction method. The approach permits detection at low concentration of trans-resveratrol. The combination of improved sample pretreatment and an isocratic chromatographic system in conjunction with internal standardization forms the basis of a new assay for the quantitation of trans-resveratrol in wine. Full-scan mass spectra were readily obtained from 8 ng of trans-resveratrol, while a limit of detection of 200 pg (signal-to-noise ratio 3) was attained in the selected ion monitoring mode. The application of LC/MS to the determination of trans-resveratrol in wines is demonstrated by the analysis of red wines. Copyright 1999 John Wiley & Sons, Ltd.

Determination of transition metals in wine by IC, DPASV-DPCSV, and ZGFAAS coupled with UV photolysis.

In the present work ion chromatography (IC), differential pulse anodic (DPASV) or cathodic stripping voltammetry (DPCSV), and Zeeman graphite furnace atomic absorption spectroscopy (ZGFAAS) were applied to the determination of heavy and transition metals in wine. The matrix is degraded by oxidative UV photolysis in a digester equipped with a high-pressure mercury lamp. The temperature of the sample is maintained at 85 +/- 5 degrees C by a combined air/water cooling system. This procedure has decisive advantages if compared with other sample pretreatment techniques. Most organic wine constituents degrade in <1 h, whereas metals quantitation remains unaffected by UV radiation, with the exception of manganese. The clear solution is directly analyzed for most common heavy and transition metals such as cadmium, cobalt, copper, iron, lead, nickel, and zinc. In the absence of standards, the results obtained by different techniques are compared and are found in good agreement. All of the considered techniques appear to be equivalent, but ZGFAAS is more time-consuming because it does not permit multielement analyses.

Relationships of Resveratrol, Anthocyanin, and Soluble Solids to Maturity in Vitis lambrusca

Resveratrol in Vitis vinifera cultivars has been shown to be important in resistance to infection of Botrytis cinerea (gray mold). Also, resveratrol is a constituent of wine that has been shown to confer protection against artherosclerosis, heart disease, and cancer. The relationship between grape maturity and resveratrol concentration levels, soluble solids, and anthocyanins were examined on V. lambrusca cultivars. `Concord' and `Catawba' grapes were harvested weekly from August to maturity from a production site in Lexington, Ky. Irradiation and non-irradiation of berries was performed in the laboratory and resveratrol concentration was analyzed. Berry skins were extracted with methanol/water (8:1 v/v) in 10 ml/g of fresh weight tissue. Resveratrol was quantified using reverse-phase HPLC. Soluble solids were measured weekly by refractometer, and anthocyanin content was determined by spectrophotometer. Results showed a marked increase in resveratrol concentration as grapes matured, and followed by a subsequent decrease at final harvest. Sugar content increased weekly in each cultivar, and was maximal at final harvest.

Isolation, identification, and antioxidant activity of three stilbene glucosides newly extracted from vitis vinifera cell cultures

Suspension cultures of Vitis vinifera L. (Vitaceae) produce many hydroxylated stilbene glucosides found in red wine. From these cells, we isolated and characterized glycosylated stilbenes, (Z)-piceatannol (3,5,3',4'-tetrahydroxystilbene) -3-O-beta-d-glucopyranoside (6) and (E)- and (Z)-resveratrol (3,5, 4'-trihydroxystilbene)-4'-O-beta-d-glucopyranoside (2 and 7, respectively), which have not previously reported to be constituents of Vitis vinifera or wine. The ability of these compounds to act as radical scavengers was investigated using 1,1 diphenyl-2-picryl-hydrazyl, a stable free radical. Antioxidant activities were assessed by their capacity to prevent Cu2+-induced lipid peroxidation in human low-density lipoprotein.

Isolation of Two Novel Terpenoid Glucose Esters from Riesling Wine

A glycosidic isolate of Riesling wine was separated with multilayer coil countercurrent chromatography (MLCCC). After acetylation and subsequent purification by high-performance liquid chromatography (HPLC), the glucose esters of (E)-2,6-dimethyl-6-hydroxyocta-2,7-dienoic acid (1) and (2E,6E)-10,11-dihydroxy-3,7,11-trimethyl-2,6-dodecadienoic acid (2) were identified for the first time as wine constituents. The identification was achieved by mass spectrometry (DCI-MS) as well as nuclear magnetic resonance spectroscopy (1H NMR, 13C NMR, COSY, HMBC). In biomimetic studies carried out with (E)-2,6-dimethyl-6-hydroxyocta-2,7-dienoic acid the potent wine aroma compound 3a,4,5,7a-tetrahydro-3,6-dimethylbenzofuran-2(3H)-one (wine lactone) was formed, thus revealing an important aroma precursor function for glucoconjugate 1. Keywords: (E)-2,6-Dimethyl-6-hydroxyocta-2,7-dienoic acid glucose ester; (2E,6E)-10,11-dihydroxy-3,7,11-trimethyl-2,6-dodecadienoic acid glucose ester; aroma precursor; wine lactone; multilay...