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The protective ability of the Mediterranean plant extracts against the oxidative DNA damage. The role of the radical oxygen species and the polyphenol content.

The polyphenol plant extracts content seems to be responsible for the scavenging activity of the reactive oxygen species (ROS), resulting in protection against DNA damage induced by the oxidative stress. This assumption was verified analyzing the effect of six Mediterranean plant extracts (Crepis vesicaria L, Origanum heracleoticum, Scandix australis L, Amaranthus sp., Scolymus hispanicus L, Thymus piperella L) on the oxidative DNA damage induced in lymphocytes by H(2)O(2) in relation to the polyphenolic content and the lymphocyte scavenging ability of ROS. The comet assay was used to evaluate oxidative DNA damage and the polyphenol content was analyzed by the Folin-Ciocalteu method. The fluorescence resulting from oxidation of ROS-sensitive dye, dihydrorofluorescein (DHF), was utilized as indicator of the ROS level. Pretreatment with all plant extracts produced the dose-dependent increase in the DNA protection up to the 0.2 microg/ml polyphenol content and the decrease above that dose. Only the Thymus piperella, similarly to quercetin, showed a strong positive correlation between the DNA protection and the polyphenol content, but negative correlation with ROS formation. In conclusion, the DNA protective ability of plant extracts seems to be related to the low polyphenol concentration and only to certain extent depends on the polyphenol ROS scavenging activity.

Comet Assay↗

Polyphenols produced during red wine ageing.

Over the past few years, it has been accepted that a moderate red wine consumption is a factor beneficial to human health. Indeed, people of France and Italy, the two major wine-producing European countries, eat a lot of fatty foods but suffer less from fatal heart strokes than people in North-America or in the northern regions of Europe, where wine is not consumed on a regular basis. For a time, ethanol was thought to be the "good" chemical species hiding behind what is known as the "French paradox". Researchers now have turned their investigations towards a family of natural substances called "polyphenols", which are only found in plants and are abundant in grapes. It is well known that these molecules behave as radical scavengers and antioxidants, and it has been demonstrated that they can protect cholesterol in the LDL species from oxidation, a process thought to be at the origin of many fatal heart attacks. However, taken one by one, it remains difficult to demonstrate which are the best polyphenols as far as their antioxidant activities are concerned. The main obstacle in that kind of research is not the design of the chemical and biological tests themselves, but surprisingly enough, the limited access to chemically pure and structurally elucidated polyphenolic compounds. In this article, particular attention will be paid to polyphenols of red wine made from Vitis vinifera cultivars. With respect to the "French paradox", we address the following question: are wine polyphenolic compounds identical to those found in grapes (skin, pulp and seed), or are there biochemical modifications specifically taking place on the native flavonoids when a wine ages? Indeed, structural changes occur during wine conservation, and one of the most studied of those changes concerns red wine colour evolution, called "wine ageing". As a wine ages, it has been demonstrated that the initially present grape pigments slowly turn into new more stable red pigments. That phenomenon goes on for weeks, months and years. Since grape and wine polyphenols are chemically distinct, their antioxidant activities cannot be the same. So, eating grapes might well lead to beneficial effects on human health, due to the variety and sometimes large amounts of their polyphenolic content. However, epidemiological surveys have focused on wines, not on grapes....

Anthocyanins↗

4-Hydroxyanisole: the most suitable monophenolic substrate for determining spectrophotometrically the monophenolase activity of polyphenol oxidase from fruits and vegetables.

A continuous spectrophotometric method for determining the monophenolase activity of polyphenol oxidase from several plant sources is described. This assay method is based on the coupling reaction between 3-methyl-2-benzothiazolinone hydrazone and the quinone product of the oxidation of 4-hydroxyanisole in the presence of polyphenol oxidase. 4-Hydroxyanisole proved to be the best monophenol assayed to measure the monophenolase activity of polyphenol oxidase from apple, artichoke, avocado, medlar, pear, and strawberry. Kinetic constants of 4-hydroxyanisole were compared to those of p-hydroxyphenyl propionic acid, a very sensitive monophenol previously reported to assay the monophenolase activity of polyphenol oxidase from apple, pear, and mushroom. The high values of the maximum steady state rate obtained for 4-hydroxyanisole suggest the existence of high catalytic constant toward this monophenol. These kinetic values were supported by nuclear magnetic resonance assays which predicted the highest reactivity of 4-hydroxyanisole. Therefore nuclear magnetic resonance assays proved to be a valuable and useful tool to predict the best monophenolic substrate for plant polyphenol oxidases. The 3-methyl-2-benzothiazlolinone-adduct for 4-hydroxyanisole was stable, with high molar absorptivity at the optimum pHs of the polyphenol oxidases assayed. All this together makes the use of 4-hydroxyanisol as monophenolic substrate and 3-methyl-2-benzothiazolinone as coupling reagent the most sensitive and precise assay method up to date reported in the literature to determine the monophenolas activity of polyphenol oxidase from fruits and vegetables.

Anisoles↗

Evaluation of polyphenol bioavailability in rat small intestine.

BACKGROUND: Dietary polyphenols, which are contained in several foods of plant origin, have been reported to be effective protective agents against cardiovascular diseases and cancer. However, data on their absorption from the gastrointestinal tract are still scarce and, often, contradictory. AIM OF THE STUDY: In this report, evaluation of polyphenol bioavailability was carried out by using segments of the small intestine from rat. The extent of absorption throughout the small intestine of rat was evaluated with two model compounds, tannic acid and catechin, as representatives of high and low molecular weight polyphenols, respectively. The consequence of the binding of tannic acid to BSA on both tannic acid absorption and in vivo protein digestibility was also examined. METHODS: Polyphenol solutions of different concentrations were injected into the lumen of ligated segments (6 cm) of the small intestine and the segments incubated in buffer for 5 min. The residual amount of polyphenol in the lumen of each segment was assayed by maximum absorption in the UV/VIS optical spectrum as was the amount of compound that had crossed the gut wall into the incubation buffer. Digestibility of BSA and of a BSA- tannic acid complex was assayed with rats. RESULTS: The results indicated a significant, concentration-dependent, disappearance of both polyphenols from the small intestine of the rat, with higher uptake levels being evident for tannic acid (50%) than catechin (30%). However, complete transfer through the gut wall was not observed with tannic acid whilst low but significant amounts (10%) were detected in incubation buffers with catechin. Partial binding of polyphenols by endogenous proteins in the intestinal lumen was also demonstrated. Complexing tannic acid with BSA (1:10 mol/mol) was not found to affect either the extent of interaction of tannic acid with the small intestine or the in vivo digestibility of the protein. CONCLUSIONS: Our experiments show that tannic acid and catechin both interact with the gut but only catechin appears able to traverse the gut. In addition, they provide evidence for binding of tannic acid and catechin by endogenous proteins in the intestinal lumen. This may limit their absorption from the small intestine. BSA complexed with tannic acid was as readily digested as BSA alone. This may suggest that tannic acid exerts anti-nutritional effects by binding to proteins of the gut wall and interfering with gut function rather than by inhibition of dietary protein digestion.

Animals↗

Potent suppressing activity of the non-polyphenolic fraction of green tea (Camellia sinensis) against genotoxin-induced umu C gene expression in Salmonella typhimurium (TA 1535/pSK 1002)--association with pheophytins a and b.

Antigenotoxic and antimutagenic activities of green tea extract and tea-derived polyphenols have been studied using in vitro and in vivo experiments. However, antigenotoxic substances in the non-polyphenolic fraction of green tea have been poorly elucidated. In the present study, the effect of the non-polyphenolic fraction of green tea on genotoxin-induced umu C gene expression was analyzed using a tester bacteria, and potent antigenotoxic substances in the non-polyphenolic fraction were identified. The non-polyphenolic fraction of green tea showed strong suppressive activities against umu C gene expression in Salmonella typhimurium (TA 1535/pSK 1002) induced by 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indol (Trp-P-1) or mitomycin C (MMC) in the presence or absence of S9 metabolizing enzyme mixture. The non-polyphenolic fraction of green tea exhibited major two-color bands in a silica gel TLC and they were identified as chlorophyll-related compounds, pheophytins a and b, judged by their specific colors, Rf values in silica gel TLC and absorption spectra. These pigments showed significant suppressive activities against umu C gene expression in tester bacteria induced by Trp-P- and MMC in a dose-dependent manner. These results suggest that the non-polyphenolic fraction of green tea contains pheophytins a and b as potent antigenotoxic substances.

Antimutagenic Agents↗

Multivariate analysis of antioxidant power and polyphenolic composition in red wines.

It has been demonstrated that the dietary intake of compounds having antioxidant activity is very important, and various chemical, biological, and electrochemical methods have been proposed to evaluate the antioxidant power of compounds such as polyphenols. Wine, although nonessential, has a high polyphenol content up to 2-3 g/L in red wines obtained by traditional maceration. The polyphenol content of wines is usually evaluated by the Folin reagent, which provides an appropriate response to the requirements of wine manufacturers. Because the presence of individual polyphenols may be evaluated by HPLC, more or less selective methods toward the various chemical classes of polyphenols have been developed. An HPLC method set up recently was applied to evaluate how individual polyphenols contributed to the overall antioxidant power (AOP) of 60 Italian red wines, trying to identify the effect that individual compounds may have on the total AOP. Application of the multivariate analysis allowed us to detect some determining compounds such as gallic acid and some flavonols. On the basis of the correlation between two traditional chemical methods, namely the total polyphenol determination by the Folin reagent and the flavanol determination by the condensation reaction with p-(dimethylamino)-cinnamaldehyde, it was shown that the use of these two merely chemical methods is well correlated (r = 0.83 and 0.87) to an AOP evaluation of red wines.

Antioxidants↗

Simultaneous determination of all polyphenols in vegetables, fruits, and teas.

Polyphenols, which have beneficial effects on health and occur ubiquitously in plant foods, are extremely diverse. We developed a method for simultaneously determining all the polyphenols in foodstuffs, using HPLC and a photodiode array to construct a library comprising retention times, spectra of aglycons, and respective calibration curves for 100 standard chemicals. The food was homogenized in liquid nitrogen, lyophilized, extracted with 90% methanol, and subjected to HPLC without hydrolysis. The recovery was 68-92%, and the variation in reproducibility ranged between 1 and 9%. The HPLC eluted polyphenols with good resolution within 95 min in the following order: simple polyphenols, catechins, anthocyanins, glycosides of flavones, flavonols, isoflavones and flavanones, their aglycons, anthraquinones, chalcones, and theaflavins. All the polyphenols in 63 vegetables, fruits, and teas were then examined in terms of content and class. The present method offers accuracy by avoiding the decomposition of polyphenols during hydrolysis, the ability to determine aglycons separately from glycosides, and information on simple polyphenol levels simultaneously.

Chromatography, High Pressure Liquid↗

Variability of the polyphenolic composition of cider apple (Malus domestica) fruits and juices.

Five French cider apple varieties were compared on the basis of their detailed polyphenol profile in the cortex and in the juices. Among the factors studied, variety was the most important variability factor in fruits, whereas polyphenol profiles showed an overall stability from one year to another, and a limited decrease of polyphenol concentration was observed during the starch regression period of fruit maturation. In juices, procyanidins remained the preponderant polyphenol class with concentrations up to 2.4 g/L even in centrifuged juices. Compared to the fruits, the average degree of polymerization of procyanidins was significantly reduced in the juice. Centrifugation of the crude juice had only minor effects on the polyphenol composition. For one variety, highly polymerized procyanidins with average degrees of polymerization of 25 were shown to be soluble in the centrifuged juice at a concentration of close to 1.2 g/L. Oxygenation of the juices during processing resulted in a significant decrease of all classes of native polyphenols. Catechins and procyanidins were particularly affected by oxidation, whereas caffeoylquinic acid was partly preserved. The transfer of polyphenols after pressing was maximal for dihydrochalcones and minimal for procyanidins with extraction yield values close to 80 and 30%, respectively.

Beverages↗

Polyphenolic profiles in eight apple cultivars using high-performance liquid chromatography (HPLC).

Polyphenolic compounds of apple may play an important role in physiologic functions related to human health. Different polyphenolics may have varied biological activities including antioxidant activity. The objective of this study was to investigate the profiles of polyphenolic compounds in different apple varieties and different parts of an apple. The total and individual polyphenolics differed significantly among the eight apple cultivars grown in Ontario, and the peels had higher concentrations than the flesh. Among the tested cultivars, Red Delicious and Northern Spy had the highest concentrations and Empire the lowest. Five major polyphenolic groups with a total of 16 identified individual compounds were found, among which the dihydroxycinnamic acid esters, phloretin glycosides, and flavan-3-ols were found in both flesh and peel, whereas quercetin glycosides were almost exclusively found in the peel. Cyanidin 3-galactoside was unique to and found only in red apple peels. In both apple peel and flesh, the predominant group of polyphenolics was the procyanidins, followed by quercetin glycosides in the peel and hydroxycinnamic acid esters in the flesh. 3-Hydroxyphloretin 2'-xyloglucoside was newly identified in apple. The results obtained in this study will further the understanding of the polyphenolic composition of apples and their roles in health-promoting physiological functions.

Anthocyanins↗

Polyphenols and glutathione synthesis regulation.

Polyphenols in food plants are a versatile group of phytochemicals with many potentially beneficial activities in terms of disease prevention. In vitro cell culture experiments have shown that polyphenols possess antioxidant properties, and it is thought that these activities account for disease-preventing effects of diets high in polyphenols. However, polyphenols may be regarded as xenobiotics by animal cells and are to some extent treated as such, ie, they interact with phase I and phase II enzyme systems. We recently showed that dietary plant polyphenols, namely, the flavonoids, modulate expression of an important enzyme in both cellular antioxidant defenses and detoxification of xenobiotics, ie, gamma-glutamylcysteine synthetase. This enzyme is rate limiting in the synthesis of the most important endogenous antioxidant in cells, glutathione. We showed in vitro that flavonoids increase expression of gamma-glutamylcysteine synthetase and, by using a unique transgenic reporter mouse strain, we showed increased expression in vivo, with a concomitant increase in the intracellular glutathione concentrations in muscles. Because glutathione is important in redox regulation of transcription factors and enzymes for signal transduction, our results suggest that polyphenol-mediated regulation of glutathione alters cellular processes. Evidently, glutathione is important in many diseases, and regulation of intracellular glutathione concentrations may be one mechanism by which diet influences disease development. The aim of this review is to discuss some of the mechanisms involved in the glutathione-mediated, endogenous, cellular antioxidant defense system, how its possible modulation by dietary polyphenols such as flavonoids may influence disease development, and how it can be studied with in vivo imaging.

Animals↗

Protective effects of green tea polyphenols and their major component, (-)-epigallocatechin-3-gallate (EGCG), on 6-hydroxydopamine-induced apoptosis in PC12 cells.

Green tea polyphenols exert a wide range of biochemical and pharmacological effects, and have been shown to possess antimutagenic and anticarcinogenic properties. Oxidative stress is involved in the pathogenesis of Parkinson's disease. However, although green tea polyphenols may be expected to inhibit the progression of Parkinson's disease on the basis of their known antioxidant activity, this has not previously been established. In the present study, we evaluated the neuroprotective effects of green tea polyphenols in the Parkinson's disease pathological cell model. The results show that the natural antioxidants have significant inhibitory effects against apoptosis induced by oxidative stress. 6-Hydroxydopamine (6-OHDA)-induced apoptosis in catecholaminergic PC12 cells was chosen as the in vitro model of Parkinson's disease in our study. Apoptotic characteristics of PC12 cells were assessed by MTT assay, flow cytometry, fluorescence microscopy and DNA fragmentation. Green tea polyphenols and their major component, EGCG at a concentration of 200 microM, exert significant protective effects against 6-OHDA-induced PC12 cell apoptosis. EGCG is more effective than the mixture of green tea polyphenols. The antioxidant function of green tea polyphenols may account for this neuroprotective effect. The present study supports the notion that green tea polyphenols have the potential to be effective as neuropreventive agents for the treatment of neurodegenerative diseases.

Animals↗

Polyphenol increases in safflower and cucumber seedlings exposed to strong visible light with limited water.

To assess effects of the environmental stress on polyphenol compounds (polyphenols) in plants, the polyphenol contents were investigated in the seedlings of safflower (Carthamus tinctrius L.) and cucumber (Cucumis sativus L.) grown under three types of growth conditions: control; light stress, irradiated with strong light in the visible wavelength range; and light/water stress, irradiated with strong visible light with a limited water supply. The total polyphenol contents and the amounts of the major polyphenols, especially luteolin 7-O-glucoside in safflower cotyledons, and luteolin 7-O-glucoside and luteolin in safflower foliage leaves, increased in response to both stresses. The polyphenol increasing effect of light/water stress was clearly observed in safflower compared to cucumber, suggesting that plants that are resistant to these stresses can accumulate substantial amounts of polyphenols compared to the plants which respond weakly to the stresses.

Asteraceae↗

[The effects of tea polyphenols on the adherence of cariogenic bacterium to the salivary acquired pellicle in vitro].

OBJECTIVE: It has been proved that tea polyphenols (TP) can inhibit the sucrose-dependent adherence of oral cariogenic bacterium to the tooth surface. This study is carried on to investigate the effects of tea polyphenols on the preliminary adherence of main cariogenic bacterium to salivary acquired pellicle and reveal the anticarious mechanisms of tea polyphenols further. METHODS: The hydroxyapatites were coated by saliva to form S-HA, an in-vitro model of experimental pellicle. Streptococcus mutans 3a3 and Actinomyces viscosus ATCC 19246 were chosen as the main cariogenic bacterium, and the microorganisms were radiolabelled by growing them in TPY culture medium supplemented with 3H-thymidine. Tea polyphenols solution was prepared, the concentrations of which were 0.5, 1.0, 2.0 and 4.0 mg/ml respectively. Two parts of inhibited adherence tests were done with TP-treated S-HA or TP-treated bacterium. In the first one, S-HA was immerged into the TP solution of different concentrations for 1 h before the attachment of bacterium. In the other, the bacterium were mixed with the TP solution for 1 h and then centrifuged and washed before bound to S-HA. The whole process was carried on in KCl buffer solution. The number of 3H-thymidine-labelled bacterium which attached to S-HA were determined by scintillation counting. RESULTS: In both the two parts of this study, tea polyphenols solution, the concentrations of which were changed from 1.0 to 4.0 mg/ml, could decrease the number of S. mutans 3a3 and A. viscosus ATCC 19246, and the inhibition rates rose with the increase of the concentrations of TP solution. But the inhibitory effects of 0.5 mg/ml TP solution were inconsistent in different tests. The results showed that 1.0-4.0 mg/ml TP solution could inhibit the attachments of S. mutans and A. viscosus to S-HA effectively. The inhibitory functions of tea polyphenols were so obvious that in some experiments, while the concentrations of TP solution were changed from 2.0 to 4.0 mg/ml, the number of 3H-thymidine-labelled bacterium which attached to S-HA was even less than that of the same bacterium which attached to naked HA. CONCLUSION: Tea polyphenols can inhibit the preliminary adherence of Streptococcus mutans and Actinomyces viscosus to S-HA effectively. TP may prevent enamel from caries by decreasing the adherence of main cariogenic bacterium to salivary acquired pellicle.

Actinomyces viscosus↗

[The effects of tea polyphenols on the adherence of cariogenic bacterium to the collagen in vitro].

OBJECTIVE: In the first phase of our study, it has been proved that tea polyphenols (TP) can inhibit the preliminary adherence of oral cariogenic bacterium to salivary acquired pellicle. This study is carried on to investigate the effects of tea polyphenols on the preliminary adherence of main cariogenic bacterium to collagen I and reveal the anticarious mechanisms of tea polyphenols further. METHODS: Tea polyphenols solution was prepared firstly, the concentrations of which were 0.5, 1.0, 2.0 and 4.0 mg/ml. An in-vitro model of experimental pellicle(C-HA) was formed by immerging the hydroxyapatites in 250 micrograms/ml collagen I solution for 1 h. Streptococcus mutans 3a3, Actinomyces viscosus ATCC 19246 and Lactobacillus ATCC 4546 were chosen as the main cariogenic bacterium and radiolabelled by growing them in TPY culture medium supplemented with 3H-thymidine. Two parts of inhibited adherence tests were done with TP-treated C-HA or TP-treated bacterium respectively. In the first one, C-HA was immerged into the TP solution of different concentrations for 1 h before the attachment of bacterium. In the other, the bacterium were mixed with the TP solution for 1 h and then centrifuged and washed before bound to C-HA. KCl buffer solution provided a steady condition to the whole process. Finally, the number of 3H-thymidine-labelled bacterium which attached to C-HA were determined by scintillation counting. RESULTS: In both the two parts of this study, tea polyphenols solution, the concentrations of which were changed from 1.0 to 4.0 mg/ml could inhibit the attachments of S. mutans 3a3, A. viscosus ATCC 19246 and Lactobacillus ATCC 4546 to C-HA effectively, and the inhibition rates rose with the increase of the concentrations of TP solution. But the inhibitory effects of 0.5 mg/ml TP solution were inconsistent in different tests. The inhibitory functions of tea polyphenols were so prominent that in some experiments, while the concentrations of TP solution were higher than 2.0 mg/ml, the number of 3H-thymidine-labelled bacterium which attached to C-HA was even less than that of the same bacterium which attached to naked HA. These results showed that TP had the remarkable inhibitory efficiency on the attachments of S. mutans, A. viscosus and Lactobacillus to C-HA. CONCLUSION: Tea polyphenols can inhibit the preliminary adherence of Streptococcus mutans, Actinomyces viscosus and Lactobacillus to C-HA effectively. TP may prevent dentine from caries by decreasing the adherence of main cariogenic bacterium to collagen.

Actinomyces viscosus↗

Induction of p57 is required for cell survival when exposed to green tea polyphenols.

Green tea polyphenols (catechins) are known to induce cell death in many types of tumor cells, but how normal epithelial cells survive in the presence of polyphenols is unknown. We recently reported that green tea polyphenols potently induced a cyclin-dependent kinase inhibitor, p57/(KIP2), only in normal human epithelial cells. In this study, we investigated the correlation between p57 expression and survival/apoptosis by Western blot analysis, caspase 3 assays and morphological analysis. It was demonstrated that, in the cells that lack p57 induction, green tea polyphenols induced Apaf-1 expression along with caspase 3 activation, leading to apoptosis. In contrast, cells with polyphenol-inducible p57 maintained constant levels of Apaf-1 and proliferating cell nuclear antigen (PCNA), with basal caspase 3 activity. Retroviral-transfected, p57-expressing oral carcinoma cells showed significant resistance to green tea polyphenol-induced apoptosis. Our results suggest that p57/KIP2 is a determinant pro-survival factor for cell protection from green tea polyphenol-induced apoptosis.

Apoptosis↗

Photochemoprevention of UVB-induced skin carcinogenesis in SKH-1 mice by brown algae polyphenols.

Chronic exposure of the skin to ultraviolet B (UVB) radiation induces oxidative stress, which plays a crucial role in the induction of skin cancer. In this study, the effect of dietary feeding and topical application of brown algae polyphenols on UVB radiation-induced skin carcinogenesis in SKH-1 mice was investigated. SKH-1 hairless mice were randomly divided into 9 groups, including control, UVB control and treatment groups. They were treated orally (0.1% and 0.5% with AIN-76 diet, w/w) and topically (3 and 6 mg/0.2 ml of vehicle) with brown algae polyphenols and irradiated with UVB for 26 weeks. Dietary feeding (0.1% and 0.5%) of brown algae polyphenols significantly reduced tumor multiplicity (45% and 56%) and tumor volume (54% and 65%), and topical administration (3 and 6 mg) significantly decreased tumor multiplicity (60% and 46%) and tumor volume (66% and 57%), respectively, per tumor-bearing mouse. Dietary feeding and topical administration of the polyphenols also inhibited tumor incidence by 6% and 21%, respectively, but the results were not significant. Dietary and topical administration of the polyphenols markedly inhibited cyclooxygenase-2 activity and cell proliferation. These observations show that brown algae polyphenols have an antiphotocarcinogenic effect which may be associated with the prevention of UVB-induced oxidative stress, inflammation, and cell proliferation in the skin.

Administration, Cutaneous↗

Dietary polyphenolic phytochemicals--promising cancer chemopreventive agents in humans? A review of their clinical properties.

Epidemiological and preclinical evidence suggests that polyphenolic phytochemicals exemplified by epigallocatechin gallate from tea, curcumin from curry and soya isoflavones possess cancer chemopreventive properties. Whilst such naturally occurring polyphenols have been the subject of numerous mechanistic studies in cells, information on their clinical properties, which might help assess their promise as human cancer chemopreventive agents, is scarce. Therefore, we present a review of pilot studies and trials with a cancer chemoprevention-related rationale, in which either healthy individuals or patients with premalignant conditions or cancer received polyphenolic phytochemicals. The review identifies trial design elements specifically applicable to polyphenolic phytochemicals. The available evidence for tea polyphenols tentatively supports their advancement into phase III clinical intervention trials aimed at the prevention of progression of prostate intraepithelial neoplasia, leukoplakia or premalignant cervical disease. In the case of curcumin and soya isoflavones more studies in premalignacies seem appropriate to optimise the nature and design of suitable phase III trials. The abundance of flavonoids and related polyphenols in the plant kingdom makes it possible that several hitherto uncharacterised agents with chemopreventive efficacy are still to be identified, which may constitute attractive alternatives to currently used chemopreventive drugs.

Antineoplastic Agents, Phytogenic↗

Ultrasonic extraction followed by a novel filtration and clean-up device for screening of some polyphenols in tobaccos.

A simple and efficient HPLC method using a diode array detector (DAD) was developed for simultaneous determination of six polyphenols in tobaccos. Ultrasonic extraction was employed at 25 degrees C to extract the polyphenols present in tobaccos into anhydrous methanol. A novel filtration device linked to a clean-up cartridge was designed for simultaneous extract filtration and clean-up. Optimized HPLC-DAD analysis, with multi-wavelength detection, was used for determination of the polyphenols. Because the content of some of the polyphenols is too low to be quantified directly, a concentration step was necessary. Anhydrous methanol was employed for extraction of the polyphenols because of its high extraction efficiency and its low boiling point in the concentration step. Using the proposed method, six polyphenols were quantified in tobaccos (Nicotina tobaccum L.).

Chromatography↗