Screening for antioxidant activity in edible plant products: comparison of low-density lipoprotein oxidation assay, DPPH radical scavenging assay, and Folin-Ciocalteu assay

Oxidation of low-density lipoprotein (LDL) has been implicated in atherogenesis. Antioxidants that prevent LDL from oxidizing may reduce atherosclerosis. This study investigated LDL antioxidant activity in edible plant products for development of dietary supplementation to prevent atherosclerosis. Fifty-two kinds of edible plants were extracted using 70% aqueous ethanol solution, and the antioxidant activity of the extracts, which inhibit human LDL oxidation induced by copper ion, was determined on the basis of the oxidation lag time and represented as epigallocatechin 3-gallate equivalent. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenolic content were also measured for comparisons with antioxidant activity in LDL. Plant products showing the greatest activity in LDL oxidation assay were akamegashiwa (Mallotus japonicus) leaf, Japanese privet (Ligustrum japonicum) leaf, green tea [Camellia sinensis (L.) O. Kuntze], and astringent persimmon (Diospyros kaki). The present study revealed high levels of LDL antioxidant activity in plant products for which such activity levels are underestimated in the DPPH radical scavenging assay and Folin-Ciocalteu assay.     

Guava fruit (Psidium guajava L.) as a new source of antioxidant dietary fiber.

Guava (Psidium guajava L.) is a tropical fruit, widely consumed fresh and also processed (beverages, syrup, ice cream, and jams). Pulp and peel fractions were tested, and both showed high content of dietary fiber (48.55-49.42%) and extractable polyphenols (2.62-7.79%). The antioxidant activity of polyphenol compounds was studied, using three complementary methods: (i) free radical DPPH* scavenging, (ii) ferric reducing antioxidant power assay (FRAP), and (iii) inhibition of copper-catalyzed in vitro human low-density lipoprotein (LDL) oxidation. All fractions tested showed a remarkable antioxidant capacity, and this activity was correlated with the corresponding total phenolic content. A 1-g (dry matter) portion of peel contained DPPH* activity, FRAP activity, and inhibition of copper-induced in vitro LDL oxidation, equivalent to 43 mg, 116 mg, and 176 mg of Trolox, respectively. These results indicate that guava could be a suitable source of natural antioxidants. Peel and pulp could also be used to obtain antioxidant dietary fiber (AODF), a new item which combines in a single natural product the properties of dietary fiber and antioxidant compounds.     

In vitro antioxidant activity and antigenotoxicity of 5-n-alkylresorcinols

Incubation with 5-n-alkylresorcinols (chain lengths C15:0, C17:0, C19:0, C21:0, and C23:0) increased the self-protection capacity of HT29 human colon cancer cells against DNA damage induced by hydrogen peroxide and genotoxic fecal water samples using comet assay (single-cell gel electrophoresis assay). The alkylresorcinols did not exert potent antioxidant activity in the FRAP (ferric reduction ability of plasma) and DPPH (2,2-diphenyl-1-picrylhydrazyl) radical assays. However they were able to significantly inhibit copper-mediated oxidation of human LDL (low-density lipoprotein) in vitro, and pentadecylresorcinol at 25 micromol/L increased lag time by 65 min. The results show that alkylresorcinols have antigenotoxic and antioxidant activity under in vitro conditions.     

Commercial dietary antioxidant supplements assayed for their antioxidant activity by different methodologies

Seven commercial dietary antioxidant supplements were evaluated for their in vitro antioxidant capacity by different methodologies: antiradical activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH.), inhibition of methyl linoleate (MeLo) autoxidation, and resistance to ion-dependent oxidation of human low-density lipoprotein (LDL). Great variability in antioxidant activity was seen among the supplements, as well as different patterns of antioxidant capacity depending on the method used. The same orders of activity were found for the MeLo autoxidation and LDL-induced oxidation methods, but the differences recorded by the former were wider. "Food/compound intake equivalents" of the doses recommended by the manufacturers were calculated in terms of quercetin, strawberries, and red wine for each of the supplements studied. Food/compound intake equivalents varied between 7.9 and 190 mg of quercetin according to the DPPH. scavenging method, between 0.20 and 98 mg according to the MeLo autoxidation method, and between 3.4 and 83 mg according to LDL-induced oxidation. In equivalent terms of red wine, the food/compound intake equivalents varied between 7 and 159 mL, between 3 and 1354 mL, and between 4 and 89 mL for the same three methods. In terms of equivalents of strawberry, they varied between 14 and 343 mg according to the DPPH. scavenging method and between 57 g and 26 kg according to the MeLo autoxidation method. These results show the need to standardize dietary supplements in terms of their antioxidant capacity to match required doses to the oxidative status of consumers.     

Artichoke (Cynara scolymus L.) byproducts as a potential source of health-promoting antioxidant phenolics

The present study reports a fast, economical, and feasible way to extract antioxidant phenolics from artichoke byproducts: raw artichoke (RA), blanched (thermally treated) artichoke (BA), and artichoke blanching waters (ABW). These byproducts represent a huge amount of discarded material in some industries. Two protocols, with possible industrial applicability, based on both methanol and water extractions were used. Phenolic contents (expressed as caffeic acid derivatives) (grams per 100 g of dry extract) were 15.4 and 9.9 for RA when extracted with methanol and water, respectively; 24.3 and 10.3 for BA when extracted with methanol and water, respectively; and finally, 11.3 g of phenolics/100 mL of ABW. Therefore, methanol extracts yielded more phenolics than water extracts, especially when BA byproducts were used. The higher amount of phenolics in BA could be due to the inactivation of polyphenol oxidase (PPO) at the industrial scale (due to blanching process), avoiding PPO-catalyzed oxidation of these phenolics, a phenomenon that could occur in RA byproducts. Artichoke extracts from industrial byproducts showed a high free radical scavenging activity (versus both DPPH* and ABTS*+ radicals) as well as capacity to inhibit lipid peroxidation (ferric thiocyanate method). According to these results, the use of artichoke extracts from industrial byproducts as possible ingredients to functionalize foodstuffs (to decrease lipid peroxidation and to increase health-promoting properties) is suggested.     

Correlation between some nutritional components and the total antioxidant capacity measured with six different assays in eight horticultural crops

The contents of antioxidant nutritional compounds, total soluble phenolics (TSP), vitamin C, vitamin E, beta-carotene, and total carotenoids (TC), were correlated with the total antioxidant capacity (AOC) of hydrophilic (HPE) and lipophilic extracts (LPE) from eight horticultural crops, namely, guava, avocado, black sapote, mango, papaya, prickly pear fruit, cladodes, and strawberry. AOC was measured using six different assays: 2,2'-diphenyl-1-picrylhydrazyl (DPPH), N,N-dimethyl-p-phenylendiamine (DMPD), ferric-ion-reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), Trolox equivalent antioxidant capacity (TEAC), and total oxidant scavenging capacity (TOSC). AOC values from HPE were about 95 times higher than LPE values. HPE of guava had the highest AOC value when evaluated with DMPD, DPPH, FRAP, TEAC, and TOSC assays, whereas with ORAC assay, black sapote had the highest value. HPE of papaya and prickly pear fruit presented the lowest AOC values with all assays. From HPE, vitamin C and TSP contents were highly correlated with AOC for all assays, while from LPE, TC and beta-carotene contents possessed a high correlation with AOC only in the DMPD assay.     

Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models.

Antioxidant-rich fractions were extracted from pomegranate (Punica granatum) peels and seeds using ethyl acetate, methanol, and water. The extracts were screened for their potential as antioxidants using various in vitro models, such as beta-carotene-linoleate and 1,1-diphenyl-2-picryl hydrazyl (DPPH) model systems. The methanol extract of peels showed 83 and 81% antioxidant activity at 50 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. Similarly, the methanol extract of seeds showed 22.6 and 23.2% antioxidant activity at 100 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. As the methanol extract of pomegranate peel showed the highest antioxidant activity among all of the extracts, it was selected for testing of its effect on lipid peroxidation, hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL) oxidation. The methanol extract showed 56, 58, and 93.7% inhibition using the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL oxidation, respectively, at 100 ppm. This is the first report on the antioxidant properties of the extracts from pomegranate peel and seeds. Owing to this property, the studies can be further extended to exploit them for their possible application for the preservation of food products as well as their use as health supplements and neutraceuticals.     

Antioxidant activity of glyceollins derived from soybean elicited with Aspergillus sojae

The extract of soybean exposed to biotic elicitors such as food-grade fungus is known to have antioxidant activity. Glyceollins were major bioactive compounds present in soybean elicited by fungi and shown to have antifungal and anticancer activities. The purpose of present study was to evaluate the antioxidant activities of glyceollins by measuring ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, singlet oxygen quenching, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, hydroxyl radical scavenging activity, and lipid peroxidation inhibition. In addition, the antioxidant potential of glyceollins were measured by a fluorescent probe, 2',7'-dichlorofluorescin diacetate (DCFDA), and dihydroethidium (DHE) in mouse hepatoma hepa1c1c7 cells in which they were insulted with H2O2 to generate reactive oxygen species (ROS). Glyceollins showed a strong reducing power and inhibited lipid peroxidation, with significant scavenging activities of radicals including singlet oxygen, superoxide anion, ABTS, and DPPH. We also found that glyceollins significantly suppressed H2O2-induced ROS production in hepa1c1c7 cells. Therefore, glyceollins deserve further study as natural antioxidants and nutraceuticals.     

Effects of Nitrogen and Sulfur on Total Phenolics and Antioxidant Activity in Two Genotypes of Leaf Mustard

Leaf mustard (Brassica juncea Coss) is widely used for both fresh and processed markets in southern China. It contains high nutritional and medicinal compounds, which are important for maintaining optimum health. The objective of this study was to determine the influence of nitrogen (N) and sulfur (S) nutrition on total phenolics and antioxidant activity in two genotypes of leaf mustard (cvs. ‘Xuelihong’ and ‘Zhujie’). Plants were greenhouse-grown using nutrient solutions with two levels of nitrogen (10 and 25 mM) and three levels of sulfur (0.5, 1, and 2 mM). Total phenolic concentrations were considerably decreased by increasing nitrogen supply, whereas increased by increasing sulfur supply. Total phenolic concentrations in cv ‘Zhujie’ was higher than in cv ‘Xuelihong’. Three assays including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, β -carotene bleaching (BCB), and ferric reducing antioxidant power (FRAP) were used to evaluate antioxidant activity. Increasing nitrogen supply reduced DPPH radical scavenging activity and FRAP value, but increased antioxidant activity using BCB assay. Increasing sulfur supply increased antioxidant activity with all three tests. The effects of genotype on DPPH radical scavenging activity and FRAP value were not significant, however, antioxidant activity using BCB assay was significantly higher in cv ‘Zhujie’ than in cv ‘Xuelihong’. A significantly positive correlation was found between DPPH radical scavenging activity and total phenolic concentrations in two genotypes, FRAP value and total phenolic concentrations in cv ‘Xuelihong’.     

Antioxidant activity of isolated ellagitannins from red raspberries and cloudberries

Ellagitannins from red raspberries (Rubus idaeus) and cloudberries (Rubus chamaemorus) were isolated by using column chromatography and preparative HPLC. The berry phenolic isolates consisted of 80% (cloudberry) and of 60% (raspberry) of ellagitannins, with raspberries also containing anthocyanins. The main ellagitannins of both raspberries and cloudberries were identified by ESI-MS to consist of the dimeric sanguiin H-6 and the trimeric lambertianin C. Monomeric ellagitannins such as casuarictin in raspberries and pedunculagin in cloudberries were also found. The antioxidant activity of the berry phenolic isolate, ellagitannin isolate (mixture), ellagitannin main fraction (dimer and trimer), and ellagic acid was studied in bulk and emulsified methyl linoleate, in human low-density lipoprotein in vitro, and the radical scavenging activity was studied in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. Cloudberry and red raspberry ellagitannins were highly effective as radical scavengers. Berry ellagitannins also showed significant antioxidant activity toward oxidation of both human LDL and methyl linoleate emulsions. However, only weak or moderate antioxidant activity was exhibited by ellagitannins toward oxidation of bulk oil. Thus, ellagitannins contribute significantly to the antioxidant capacity of cloudberries and red raspberries in lipoprotein and lipid emulsion environments, the latter being more relevant for food applications.     

Chemical composition and hepatoprotective effects of polyphenol-rich extract from Houttuynia cordata tea

This study was designed to investigate the antioxidant activity, hepatoprotective effect, and phenolic composition of the ethyl acetate fraction (EAF) extracted from Houttuynia cordata tea. EAF was shown to exhibit strong ferric-reducing antioxidant power (FRAP) and scavenging activity against DPPH radical in vitro, and the antioxidant effects were further verified by suppressing CCl?-induced oxidative stress in mouse liver at three tested doses of EAF (250, 500, and 1000 mg/kg bw). Pretreatment with EAF (1000 mg/kg bw) prior to CCl? administration significantly (p < 0.001) decreased the CCl?-elevated levels of serum AST, ALT, alkaline phosphatase, total bilirubin, and hepatic MDA in mice and prevented the increases in GSH, SOD, and CAT caused by CCl?. HPLC analysis revealed that three predominantly polyphenolic compounds present in EAF were quercitrin (111.7 μg/mg), quercetin (43.8 μg/mg), and hyperoside (29.1 μg/mg). These results combined with liver histopathology indicate that EAF possesses a significant protective effect against acute hepatotoxicity induced by CCl?, which may be due to the strong antioxidant activity of phenolic components.     

Effects of extraction methods on phenolic contents and antioxidant activity in aerial parts of Potentilla atrosanguinea Lodd. and quantification of its phenolic constituents by RP-HPLC

The effects of different solvent systems (methanol, ethanol, acetone, and their 50% aqueous concentrations) and extraction procedures (microwave, ultrasound, Soxhlet and maceration) on the antioxidant activity of aerial parts of Potentilla atrosanguinea were investigated by three different bioassays: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays and ferric reducing antioxidant potential (FRAP). The 50% aqueous ethanol extracts exhibited strong antioxidant activity measured in terms of Trolox equivalent antioxidant capacity (TEAC) [(54.34 to 122.96, 29.82 to 101.22 and 13.64 to 41.43) mg of Trolox/g] with ABTS (*+), DPPH (*) and FRAP assays, respectively. In general, TEAC of Soxhlet extracts was found to be 1.8 and 3 times higher than ultrasound and maceration but slightly (1.2 times) higher than microwave. A positive correlation (r(2) = 0.931 to 0.982) was observed between total polyphenol (TPC) and total flavonoid (TFC) contents which ranged between 26.7 to 30.7 mg/g gallic acid equivalent and 16.8 to 20.8 mg/g quercetin equivalent respectively, with antioxidant activity. In addition, some of its bioactive phenolic constituents which contribute largely toward antioxidant potential such as chlorogenic acid, catechin, caffeic acid, p-coumaric acid and quercetin were also quantified in different extracts by RP-HPLC.     

Novel low-density lipoprotein (LDL) oxidation model: antioxidant capacity for the inhibition of LDL oxidation

A novel model of peroxyl radical initiated low-density lipoprotein (LDL) oxidation (LDL oxidation model for antioxidant capacity, or LOMAC) was developed to assess the free radical scavenging capacity of antioxidants and the extracts of natural products. A water-soluble free radical initiator, 2,2'-azobis(amidinopropane) dihydrochloride, was used at physiological temperature (37 degrees C) to generate peroxyl radicals to catalyze lipid oxidation of LDL isolated from human plasma samples. Headspace hexanal, a major decomposition product of LDL oxidation, was measured by a headspace gas chromatograph as an indicator of antioxidant capacity of different concentrations of pure antioxidants (vitamins C and E) and the extracts of natural products (fresh apple phytochemical extracts). All vitamin C and E and apple extract concentrations tested resulted in increasing partial suppression and delay of LDL oxidation. On the basis of the median effective dose (EC(50)) calculated for each compound or extract tested, the LOMAC value of 100 g of apple against LDL oxidation was equivalent to 1470 mg of vitamin E or to 402 mg of vitamin C. This study shows that the LOMAC assay can be routinely used to analyze or screen antioxidants or phytochemical extracts against LDL oxidation to prevent cardiovascular disease. The food-specific LOMAC values will be very useful as a new alternative biomarker for future epidemiological studies of cardiovascular disease.     

Antioxidant capacity of lettuce leaf tissue increases after wounding

Wounding induced the accumulation of phenolic compounds in Iceberg and Romaine lettuce leaf tissue. Phenolic concentrations were quantified after holding the leaf tissue at 10 degrees C for 48 h as the absorbance of a methanol extract at 320 nm, and by the Folin-Ciocalteu method. Heat-shock treatments applied by immersing tissue in 45 degrees C water for 2.5 min before or after wounding reduced the accumulation of phenolic compounds. Compared to the nonwounded, nonheat-shocked controls, these and other wounding and heat-shock treatments produced leaf tissue with a 4-fold range in phenolic content. The antioxidant capacity of the tissue, measured as DPPH (alpha,alpha-diphenyl-beta-picrylhydrazyl)-radical scavenging activity, or as ferric-reducing antioxidant power (FRAP), increased after wounding. The increase was linearly correlated with the increase in phenolic compounds in Iceberg (R(2) > 0.97) and in Romaine (R(2) > 0.95) lettuce leaf tissue. Increased consumption of diets rich in phenolic antioxidants may contribute to reducing human diseases. Treatments that reduce the browning of wounded lettuce leaf tissue by preventing the oxidation of the accumulated wound-induced phenolic compounds may produce a healthier fresh-cut product than treatments that prevent the wound-induced synthesis and accumulation of phenolic compounds with antioxidant properties.     

Comparison of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States

A number of different beverage products claim to have antioxidant potency due to their perceived high content of polyphenols. Basic and applied research indicates that pomegranate juice (PJ), produced from the Wonderful variety of Punica granatum fruits, has strong antioxidant activity and related health benefits. Although consumers are familiar with the concept of free radicals and antioxidants, they are often misled by claims of superior antioxidant activity of different beverages, which are usually based only on testing of a limited spectrum of antioxidant activities. There is no available direct comparison of PJ's antioxidant activity to those of other widely available polyphenol-rich beverage products using a comprehensive variety of antioxidant tests. The present study applied (1) four tests of antioxidant potency [Trolox equivalent antioxidant capacity (TEAC), total oxygen radical absorbance capacity (ORAC), free radical scavenging capacity by 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP)]; (2) a test of antioxidant functionality, that is, inhibition of low-density lipoprotein (LDL) oxidation by peroxides and malondialdehyde methods; and (3) evaluation of the total polyphenol content [by gallic acid equivalents (GAEs)] of polyphenol-rich beverages in the marketplace. The beverages included several different brands as follows: apple juice (3), a?aí juice (3), black cherry juice (3), blueberry juice (3), cranberry juice (3), Concord grape juice (3), orange juice (3), red wines (3), iced tea beverages (10) [black tea (3), green tea (4), white tea (3)], and a major PJ available in the U.S. market. An overall antioxidant potency composite index was calculated by assigning each test equal weight. PJ had the greatest antioxidant potency composite index among the beverages tested and was at least 20% greater than any of the other beverages tested. Antioxidant potency, ability to inhibit LDL oxidation, and total polyphenol content were consistent in classifying the antioxidant capacity of the polyphenol-rich beverages in the following order: PJ>red wine>Concord grape juice>blueberry juice>black cherry juice, a?aí juice, cranberry juice>orange juice, iced tea beverages, apple juice. Although in vitro antioxidant potency does not prove in vivo biological activity, there is also consistent clinical evidence of antioxidant potency for the most potent beverages including both PJ and red wine.     

Total antioxidant capacity of arginine-conjugated linoleic acid (CLA) complex

Conjugated linoleic acids (CLA) have shown diverse biological activities, including anti-carcinogenic, anti-atherosclerotic, anti-adipogenic, and anti-diabetogenic effects. Recent reports also showed that CLA has free radical scavenging capacity, which may give health benefits to human beings. However, the application of CLA as a bioactive ingredient has been limited due to its insolubility in water. To overcome this problem, we investigated antioxidant activities of arginine (Arg)-CLA, a water-soluble CLA salt, using both 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. CLA, Arg, and Arg-CLA all exerted radical scavenging activities in a dose-dependent manner in both assays. Arg-CLA at 20 mM scavenged 89 and 55% of ABTS and DPPH radicals in 3 h, respectively, whereas CLA alone quenched only 48 and 26% of them under the same conditions. The antioxidant activity of the Arg-CLA complex was found to be synergistic in ABTS assay and comparable to that of vitamin E in DPPH assay.     

Profiles of carotenoids, anthocyanins, phenolics, and antioxidant activity of selected color waxy corn grains during maturation

Waxy corns are becoming increasingly consumed as fresh foods or as raw materials for whole grain foods facilitating human consumption in China, so they are usually harvested before complete maturity. Unfortunately, information on functional properties of immature waxy corns is very limited. Therefore, we investigated the profiles of carotenoids, anthocyanins, phenolics, and the antioxidant activity in three types of waxy corn with different colors (white, yellow, and black) during maturation, as well as a normal corn (yellow) used as control. The results showed that black waxy corn had the highest quantity of anthocyanins, phenolics and the best antioxidant activity, yellow corn contained a relatively large amount of carotenoids, while white corn had the lowest amounts of carotenoids, anthocyanins, phenolics, and antioxidant capacity. For each type of waxy corn, the higher carotenoids were found at the M2 stage (no major difference between the M1 and M2 stages for yellow corn). The levels of anthocyanin and phenolics decreased for white and yellow corns, contrary to those for black corn during maturation. The antioxidant activity determined by scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH), the ferric reducing antioxidant power (FRAP), and the Trolox equivalent antioxidant capacity (TEAC) assays increased with ripening, but no difference was found between the M2 and maturity stages for yellow and black corns. For white corn, the DPPH radical scavenging activity first increased and then decreased, while the antioxidant activity determined by TEAC and FRAP assay decreased during maturation. Differences in these parameters indicate that types and harvesting time have significant influences on functional properties of waxy corns.     

Analysis of flavonoids and other phenolic compounds using high-performance liquid chromatography with coulometric array detection: relationship to antioxidant activity

High-performance liquid chromatography coupled with a coulometric array detector was used to characterize the electrochemical behavior of 17 flavonoids and three cinnamic acid derivatives. The antioxidant activity of these phenolic compounds was evaluated by the ferric reducing activity power (FRAP), the oxygen radical absorbance capacity (ORAC), and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical assays. All flavonoids, except kaempferol-3-rutinoside, malvidin-3-glucoside, and peonidin-3-glucoside, had two oxidation potentials (100-300 and 700-800 mV). Quercetin and myricetin had an additional oxidation wave at 400 mV. The electrochemical responses at a relatively low oxidation potential (300 mV) and the cumulative responses at medium oxidation potentials (400 and 500 mV) had the highest correlations with antioxidant activities. The highest correlations between electrochemical characteristics and antioxidant activities were found between electrochemical responses and antioxidant activities obtained in the FRAP assay and in the DPPH assay after short reaction periods. Lower correlations were revealed between electrochemical responses and antioxidant activities obtained in the ORAC assay.     

Characterization of antioxidants present in bitter tea (Ligustrum pedunculare)

The present study examined the antioxidants present in bitter tea (Ligustrum pedunculare). It was found that the crude glycoside fraction strongly protected human low-density lipoprotein (LDL) from oxidation. Further column chromatography led to purification of eight phenylethanoid or monoterpene glycosides: lipedoside A-I, lipedoside A-II, lipedoside B-I, lipedoside B-III, lipedoside B-V, lipedoside B-VI, osmanthuside B, and anatolioside. It was found that lipedoside A-I, lipedoside A-II, lipedoside B-V, and lipedoside B-VI were protective, whereas the other four compounds did not protect human LDL from Cu(2+)-medicated oxidation. Lipedoside A-I, lipedoside A-II, lipedoside B-V, and lipedoside B-VI also had a scavenging effect on 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH), comparable to that of alpha-tocopherol. The inhibitory effect of these four phenylethanoid or monoterpene glycosides on oxidation of human LDL and alpha-tocopherol was dose-dependent at concentrations of 5-40 microM. The present results demonstrate that bitter tea as a beverage contains effective antioxidants that may have benefits similar to those of green tea in terms of antioxidant activity.     

Antihyperlipidemic and antioxidant effects of extracts from Pleurotus citrinopileatus

Pleurotus citrinopileatus is a popular edible mushroom which is physiologically active in both humans and animals. In the study we investigate the effects of this mushroom on hyperlipidemic hamster rats. Four dietary forms of the mushroom were created as follows. The powdered dry fruiting body, hot-water extract, and two kinds of elutes were obtained, from ethyl acetate extract and methanol extract, respectively, in different mixed proportion solvents over silica gel column chromatography (referred to as EAE and MOE, respectively). They were tested at different dosages as a supplement to a high-fat diet in hyperlipidemic rats. Serum triglycerides and total cholesterol levels were significantly lower in groups supplemented with the highest dosages of EAE and MOE (0.5 g/kg, body weight daily) as compared with the control groups that received no mushroom additive. High-density lipoprotein levels in these same two experimental groups were also significantly higher than those in the negative control group. The tested rats that were fed with EAE had the highest serum glutathione peroxidase and superoxide dismutase activity, and those with the MOE and EAE had the highest DPPH free radical scavenging activities and ferric-reducing abilities, tested in vitro. The major constituents of MOE and EAE were identified as ergosterol and nicotinic acid, respectively. P. citrinopileatus extracts may have a significant antihyperlipidemia effect. Furthermore, antioxidant activities and antihyperlipidemic effects of MOE and EAE seemed to display similar tendencies.