Isolation and structure of whiskey polyphenols produced by oxidation of oak wood ellagitannins

Three new phenolic compounds named whiskey tannins A and B and carboxyl ellagic acid were isolated from commercial Japanese whiskey, along with gallic acid, ellagic acid, brevifolin carboxylic acid, three galloyl glucoses, a galloyl ester of phenolic glucoside, 2,3-(S)-hexahydroxydiphenoylglucose, and castacrenin B. Whiskey tannins A and B were oxidation products of a major oak wood ellagitannin, castalagin, in which the pyrogallol ring at the glucose C-1 position of castalagin was oxidized to a cyclopentenone moiety. These tannins originated from ellagitannins contained in the oak wood used for barrel production; however, the original oak wood ellagitannins were not detected in the whiskey. To examine whether the whiskey tannins were produced during the charring process of barrel production, pyrolysis products of castalagin were investigated. Dehydrocastalagin and a new phenolcarboxylic acid trislactone having an isocoumarin structure were isolated, along with castacrenin F and ellagic acid. However, whiskey tannins were not detected in the products.     

Understanding the native Californian diet: Identification of condensed and hydrolyzable tannins in tanoak acorns (Lithocarpus densiflorus)

The tanoak (Lithocarpus densiflorus) acorn was a staple food in the Native American diet and is still used in traditional dishes. Acorns from the genus Quercus have been shown to contain a large range of hydrolyzable tannins. However, neither hydrolyzable nor condensed tannins have been characterized in tanoak acorns. The aim of this study was to identify the full range of hydrolyzable and condensed tannins in extracts of tanoak acorns using liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry. Condensed tannins were identified as B type oligomers of (epi)-catechin (procyanidins) with a degree of polymerization up to six. Oligomers up to and including tetramers were identified by UV spectra and MS detection whereas pentamers and hexamers were detected only by MS. The total concentration of condensed tannins was 464 mg/100 g acorn pericarp. The concentration of propocyanidin monomers, dimers, trimers, and tetramers in acorn pericarp (mg/100 g acorn pericarp) were 95 +/- 10.9, 148 +/- 35.0, 90 +/- 17.9, and 131 +/- 1.9, respectively. No procyanidins were found in the acorn cotyledon tissue. A total of 22 hydrolyzable tannins were identified in methanolic extracts of acorn cotyledon tissue. Gallic acid derivatives predominated and included galloylated esters of glucose, hexahydrodiphenoyl esters of glucose, and methylated gallates. Galloylated esters of glucose were present as isomers of galloyl glucose, digalloyl glucose, and trigalloyl glucose. Mass spectral fragmentation patterns indicate the presence of one gallic acid-galloyl glucose isomer and two gallic acid-digalloyl-glucose isomers. No isomers of tetragalloyl glucose and pentagalloyl glucose were identified. Ellagic acid and ellagic acid pentoside were also identified.     

Phenolic compounds and antioxidant activity of kernels and shells of Mexican pecan (Carya illinoinensis)

The phenolic composition and antioxidant activity of pecan kernels and shells cultivated in three regions of the state of Chihuahua, Mexico, were analyzed. High concentrations of total extractable phenolics, flavonoids, and proanthocyanidins were found in kernels, and 5-20-fold higher concentrations were found in shells. Their concentrations were significantly affected by the growing region. Antioxidant activity was evaluated by ORAC, DPPH?, HO?, and ABTS?-- scavenging (TAC) methods. Antioxidant activity was strongly correlated with the concentrations of phenolic compounds. A strong correlation existed among the results obtained using these four methods. Five individual phenolic compounds were positively identified and quantified in kernels: ellagic, gallic, protocatechuic, and p-hydroxybenzoic acids and catechin. Only ellagic and gallic acids could be identified in shells. Seven phenolic compounds were tentatively identified in kernels by means of MS and UV spectral comparison, namely, protocatechuic aldehyde, (epi)gallocatechin, one gallic acid-glucose conjugate, three ellagic acid derivatives, and valoneic acid dilactone.     

Major flavonoids in grape seeds and skins: antioxidant capacity of catechin, epicatechin, and gallic acid

Grape seeds and skins are good sources of phytochemicals such as gallic acid, catechin, and epicatechin and are suitable raw materials for the production of antioxidative dietary supplements. The differences in levels of the major monomeric flavanols and phenolic acids in seeds and skins from grapes of Vitis vinifera varieties Merlot and Chardonnay and in seeds from grapes of Vitis rotundifolia variety Muscadine were determined, and the antioxidant activities of these components were assessed. The contribution of the major monomeric flavonols and phenolic acid to the total antioxidant capacity of grape seeds and skins was also determined. Gallic acid, monomeric catechin, and epicatechin concentrations were 99, 12, and 96 mg/100 g of dry matter (dm) in Muscadine seeds, 15, 358, and 421 mg/100 g of dm in Chardonnay seeds, and 10, 127, and 115 mg/100 g of dm in Merlot seeds, respectively. Concentrations of these three compounds were lower in winery byproduct grape skins than in seeds. These three major phenolic constituents of grape seeds contributed <26% to the antioxidant capacity measured as ORAC on the basis of the corrected concentrations of gallic acid, catechin, and epicatechin in grape byproducts. Peroxyl radical scavenging activities of phenolics present in grape seeds or skins in decreasing order were resveratrol > catechin > epicatechin = gallocatechin > gallic acid = ellagic acid. The results indicated that dimeric, trimeric, oligomeric, or polymeric procyanidins account for most of the superior antioxidant capacity of grape seeds.     

Antioxidant phytochemicals in hazelnut kernel (Corylus avellana L.) and hazelnut byproducts

Antioxidant efficacies of ethanol extracts of defatted raw hazelnut kernel and hazelnut byproducts (skin, hard shell, green leafy cover, and tree leaf) were evaluated by monitoring total antioxidant activity (TAA) and free-radical scavenging activity tests [hydrogen peroxide, superoxide radical, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical], together with antioxidant activity in a beta-carotene-linoleate model system, inhibition of oxidation of human low-density lipoprotein (LDL) cholesterol, and inhibition of strand breaking of supercoiled deoxyribonucleic acid (DNA). In addition, yield, content of phenolics, and phenolic acid profiles (free and esterified fractions) were also examined. Generally, extracts of hazelnut byproducts (skin, hard shell, green leafy cover, and tree leaf) exhibited stronger activities than hazelnut kernel at all concentrations tested. Hazelnut extracts examined showed different antioxidative efficacies, expected to be related to the presence of phenolic compounds. Among samples, extracts of hazelnut skin, in general, showed superior antioxidative efficacy and higher phenolic content as compared to other extracts. Five phenolic acids (gallic acid, caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid) were tentatively identified and quantified (both free and esterified forms). Extracts contained different levels of phenolic acids. These results suggest that hazelnut byproducts could potentially be considered as an excellent and readily available source of natural antioxidants.     

Fruit maturity and juice extraction influences ellagic acid derivatives and other antioxidant polyphenolics in muscadine grapes

Polyphenolic compounds including ellagic acid, ellagic acid derivatives, and anthocyanins were characterized and quantified by novel chromatographic conditions in eight muscadine grape (Vitis rotundifolia) cultivars and evaluated for antioxidant capacity as influenced by two ripening stages and their location within the fruit (skin, pulp, and juice). All polyphenolics generally increased as fruit ripened and the highest concentrations were located in the skins. Free ellagic acid, ellagic acid glycosides, and total ellagic acid ranged from 8 to 162, 7 to 115, and 587 to 1900 mg/kg, respectively, in the skin of ripe grapes. Hot-pressed juices contained considerably lower polyphenolic concentrations than were present in whole grapes. Five anthocyanidins were present in each cultivar in variable concentrations (delphinidin > petunidin > malvidin + peonidin > cyanidin). Antioxidant capacity was appreciably influenced by cultivar, maturity, and location in the fruit with good correlations to soluble phenolics found in both methanolic and ethyl acetate extracts (r = 0.83 and 0.92, respectively).     

Lipid oxidation in emulsions as affected by charge status of antioxidants and emulsion droplets

The influence of charge status of both lipid emulsion droplets and phenolic antioxidants on lipid oxidation rates was evaluated using anionic sodium dodecyl sulfate (SDS) and nonionic polyoxyethylene 10 lauryl ether (Brij)-stabilized emulsion droplets and the structurally similar phenolic antioxidants gallamide, methyl gallate, and gallic acid. In nonionic, Brij-stabilized salmon oil emulsions at pH 7.0, gallyol derivatives (5 and 500 microM) inhibited lipid oxidation with methyl gallate > gallamide > gallic acid. In the Brij-stabilized salmon oil emulsions at pH 3.0, low concentrations of the galloyl derivatives were prooxidative or ineffective while high concentrations were antioxidative. In SDS-stabilized salmon oil emulsions, oxidation rates were faster and the galloyl derivatives were less effective compared to the Brij-stabilized emulsions. Differences in antioxidant activity were related to differences in the ability of the galloyl derivatives to partition into emulsion droplets and to increase the prooxidant activity of iron at low pH.     

Isolation of nematicidal compounds from Tagetes patula L. yellow flowers: structure-activity relationship studies against cyst nematode Heterodera zeae infective stage larvae

Bioassay-guided isolation studies on the extracts of yellow flowers of Tagetes patula L. against the Heterodera zeae were carried out to identify phytochemicals lethal to this economically important cyst nematode. In vitro investigation of a polar extract and fractions showing activity led to the isolation of phenolic compounds (flavonoids and phenolic acids). In the nonpolar extract, a few fatty acids, their methyl esters, and thiophenes (including α-terthienyl) were detected. In studies of compounds obtained commercially, α-terthienyl and gallic and linoleic acids showed 100% mortality at concentrations of 0.125% after 24 h. Assessment of structure-activity relationships revealed that an increase in the number of hydroxyl groups in phenolic acids increased the activity; with fatty acids, activity depended on chain length and the number and position of double bonds. Crude extracts of the flowers of different colors also have promising activity.     

Effect of toasting intensity at cooperage on phenolic compounds in acacia (Robinia pseudoacacia) heartwood

The phenolic composition of heartwood from Robinia pseudoacacia, commonly known as false acacia, before and after toasting in cooperage was studied by HPLC-DAD and HPLC-DAD/ESI-MS/MS. A total of 41 flavonoid and nonflavonoid compounds were identified, some tentatively, and quantified. Seasoned acacia wood showed high concentrations of flavonoid and low levels of nonflavonoid compounds, the main compounds being the dihydroflavonols dihydrorobinetin, fustin, tetrahydroxy, and trihydroxymethoxy dihydroflavonol, the flavonol robinetin, the flavanones robtin and butin, and a leucorobinetinidin, none of which are found in oak wood. The low molecular weight (LMW) phenolic compounds present also differed from those found in oak, since compounds with a β-resorcylic structure, gallic related compounds, protocatechuic aldehyde, and some hydroxycinnamic compounds are included, but only a little gallic and ellagic acid. Toasting changed the chromatographic profiles of extracts spectacularly. Thus, the toasted acacia wood contributed flavonoids and condensed tannins (prorobinetin type) in inverse proportion to toasting intensity, while LMW phenolic compounds were directly proportional to toasting intensity, except for gallic and ellagic acid and related compounds. Even though toasting reduced differences between oak and acacia, particular characteristics of this wood must be taken into account when considering its use in cooperage: the presence of flavonoids and compounds with β-resorcylic structure and the absence of hydrolyzable tannins.     

Determination of free and total phenolic acids in plant-derived foods by HPLC with diode-array detection

A high-performance liquid chromatographic (HPLC) method with diode-array detection (DAD) was used to identify and quantify free and total phenolic acids (m-hydroxybenzoic acid, p-hydroxybenzoic acid, protocatechuic acid, gallic acid, vanillic acid, syringic acid, o-coumaric acid, m-coumaric acid, p-coumaric acid, caffeic acid, ferulic acid, sinapic acid, chlorogenic acid, and ellagic acid) in plant foods. Free phenolic acids were extracted with a mixture of methanol and 10% acetic acid. Bound phenolic acids were liberated using first alkaline and then acid hydrolysis followed by extraction with diethyl ether/ethyl acetate (1:1). All fractions were quantified separately by HPLC. After HPLC quantification, results of alkali and acid hydrolysates were calculated to represent total phenolic acids. Ellagic acid was quantified separately after long (20 h) acid hydrolysis. The methods developed were effective for the determination of phenolic acids in plant foods. DAD response was linear for all phenolic acids within the ranges evaluated, with correlation coefficients exceeding 0.999. Coefficients of variation for 4-8 sample replicates were consistently below 10%. Recovery tests of phenolic acids were performed for every hydrolysis condition using several samples. Recoveries were generally good (mean >90%) with the exceptions of gallic acid and, in some cases, caffeic acid samples.     

Phenolic content and antioxidant capacity of muscadine grapes

Fruits of 10 cultivars of muscadine grapes (five bronze skin and five purple skin) grown in southern Georgia were separated into skin, seed, and pulp. Each fruit part and the leaves from the corresponding varieties were extracted for HPLC analysis of major phenolics. Total phenolics were determined colorimetrically using Folin-Ciocalteu reagent. Total anthocyanins were determined according to a pH-differential method, using a UV-visible spectrophotometer. Antioxidant capacity was determined by the Trolox equivalent antioxidant capacity (TEAC) assay. Gallic acid, (+)-catechin, and epicatechin were the major phenolics in seeds, with average values of 6.9, 558.4, and 1299.4 mg/100 g of fresh weight (FW), respectively. In the skins, ellagic acid, myricetin, quercetin, kaempferol, and trans-resveratrol were the major phenolics, with respective average values of 16.5, 8.4, 1.8, 0.6, and 0.1 mg/100 g of FW. Contrary to previous results, ellagic acid and not resveratrol was the major phenolic in muscadine grapes. The HPLC solvent system used coupled with fluorescence detection allowed separation of ellagic acid from resveratrol and detection of resveratrol. Reported here for the first time are the phenolic content and antioxidant capacity of muscadine leaves. Major phenolics in muscadine leaves were myricetin, ellagic acid, kaempferol, quercetin, and gallic acid, with average concentrations of 157.6, 66.7, 8.9, 9.8, and 8.6, respectively. Average total phenolics were 2178.8, 374.6, 23.8, and 351.6 mg/g gallic acid equivalent in seed, skin, pulp, and leaves, respectively. Total anthocyanin contents were 2.1 and 132.1 mg/100 g of FW in the skins of bronze and purple grapes, respectively, and 4.3 and 4.6 mg/100 g of FW in seeds and pulps, in that order. Antioxidant capacity values were, on average, 2.4, 12.8, 281.3, and 236.1 microM TEAC/g of FW for pulps, skins, seeds, and leaves, respectively.     

Effect of tannins from Quercus suber and Quercus coccifera leaves on ethanol-induced gastric lesions in mice

The gastroprotective effects of 70% acetone extracts of Quercus suber and Quercus coccifera leaves and of tannins (pedunculagin, castalagin, phillyraeoidin A, and acutissimin B) purified from these extracts were examined in the mouse using the ethanol-induced gastric ulcer model. Both extracts (25, 50, and 100 mg/kg), given orally, prevented the formation of ethanol-induced lesions in the stomach. The percent protection varied between 68 and 91%. Purified tannins (50 mg/kg) were also effective in protecting the stomach against ethanol, and the percent protection varied from 66 to 83%. Castalagin was the most potent. Both extracts and all of the tannins tested (10, 25, and 50 microg/mL) strongly inhibited (55-65%) the lipid peroxidation of rabbit brain homogenate. These results suggest that the gastroprotective effects of extracts of Q. suber and Q. coccifera leaves and the purified tannins in this experimental model are related to their anti-lipoperoxidant properties.     

Identification and quantification of phenolic compounds in berries of Fragaria and Rubus species (family Rosaceae)

High-performance liquid chromatography combined with diode array and electrospray ionization mass spectrometric detection was used to study soluble and insoluble forms of phenolic compounds in strawberries, raspberries (red and yellow cultivated and red wild), arctic bramble, and cloudberries. Hydroxycinnamic acids were present as free forms in cloudberries and mainly as sugar esters in the other berries. Quercetin 3-glucuronide was the typical flavonol glycoside in all of the berries studied. The composition of the predominant anthocyanins can be used to distinguish the studied red Rubus species from each other since cyanidin was glycosylated typically with 3-sophorose (56%) in cultivated red raspberry, with 3-sophorose (30%) and 3-glucose (27%) in wild red raspberry, and with 3-rutinose (80%) in arctic bramble. Ellagic acid was present as free and glycosylated forms and as ellagitannins of varying degrees of polymerization. Comparable levels of ellagitannins were obtained by the analysis of soluble ellagitannins as gallic acid equivalents and by the analysis of ellagic acid equivalents released by acid hydrolysis of the extracts.     

四种栎树EST-SSR信息分析

为开发栎属遗传多样性检测的SSR标记,分析了蒙古栎、无梗花栎、夏栎和欧洲栓皮栎EST-SSR的特点,结果表明蒙古栎的EST中3 209.46 bp有一个SSR,无梗花栎中每6 160.36 bp有一个SSR、夏栎中每5 883.30 bp有一个SSR,欧洲栓皮栎中每6 129.12 bp有一个SSR。蒙古栎、无梗花栎、夏栎和欧洲栓皮栎EST-SSR的平均长度分别为21.65 bp、21.1 bp、20.66 bp和20.65 bp。四种栎类中不同基元的EST-SSR的分布频率具有非常一致的特征,均是二基元、三基元和六基元的SSR分布频率最高,达20%以上。而四基元和五基元的SSR在四个种类中的分布不到0.05%。二基元的SSR中大于1%的SSR均是AG、CT、TC、GA、AT、TA基元,并且在蒙古栎、无梗花栎和夏栎中AG、CT、TC基元的分布频率最高,而在欧洲栓皮栎中是TC、GA、AG的分布频率最高;三基元的SSR中,含CAA、GAA、TCT、CTT的SSR在四种栎类中都存在。六基元的SSR中大于1%在四种栎类中出现的类型均较少,为0～4种。     

Gallic esters of sucrose as efficient radical scavengers in lipid peroxidation

Three tests of increasing complexity were used to assess the antioxidant activity of five synthetic gallic esters of sucrose bearing 3, 6, 7, or 8 galloyl units. In addition, two of these compounds had 1 or 2 hydrocarbon (C10-C12) acyl chains. Reaction with the DPPH radical led to the evaluation of the number of radicals trapped per galloyl unit n (3-4), as well as the apparent second-order rate constant for H atom donation k (1200-1500/M/s). These results indicated similar contribution and reactivity of all the galloyl units. Inhibition of the AAPH-initiated peroxidation of linoleic acid in a micellar medium confirmed the additive contribution of the galloyl units, whereas the presence of the hydrocarbon acyl chains had no influence. These results suggest an inhibition of initiation at high antioxidant levels and an underlying prooxidant effect of the galloyl radicals at low concentrations. Finally, LDL peroxidation was inhibited in proportion to the number of galloyl units, in agreement with the preceding tests.     

RP-HPLC analysis of the phenolic compounds of plant extracts. investigation of their antioxidant capacity and antimicrobial activity

Extracts of aromatic plants of Greek origin were examined as potential sources of phenolic compounds. RP-HPLC with UV detection was employed for the identification and quantification of the phenolic antioxidants, present in methanolic extracts. The most abundant phenolic acids were ferulic acid (1.1-280 mg/100 g of dry sample) and caffeic acid (1.2-60 mg/100 g of dry sample). (+)-Catechin and quercetin were the most abundant flavonoids. Apigenin and luteolin were detected in high amounts in Menta pulegium and Thymus vulgaris, respectively. The antioxidant capacity was determined, in dried ground plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically according to the Folin-Ciocalteu assay and ranged from 1 to 21 mg of gallic acid/100 g of dry sample. Antimicrobial activity of the extracts against selected microbes was also conducted in this study.     

Phenolic composition of champagnes from Chardonnay and Pinot Noir vintages

Nineteen phenolic compounds including hydroxybenzoic acids, hydroxycinnamic acids, flavonoids, phenolic alcohols, and phenolic aldehydes have been identified and quantified in two monovarietal champagnes, Chardonnay and Pinot Noir, by using a reverse-phase high-performance liquid chromatography (HPLC) system coupled with diode array detection. The identification of four hydroxycinnamic tartaric esters (caftaric, coutaric, fertaric, and 2-S-glutathionylcaftaric acids), two flavanonols (astilbin and engeletin), and some other compounds was confirmed by HPLC coupled with mass spectrometry. Caftaric acid and tyrosol were the major phenols. Hydroxybenzoic acids and flavonoids were present at low concentrations. The phenolic compositions of 2000 and 2001 Chardonnay and Pinot Noir vary quantitatively according to the year and the variety, but the chemical natures of the molecules are the same. The total phenolic content determined by colorimetric measurement ranges from 176 to 195 mg/L of gallic acid equivalent and is similar to that described in white wines.     

Analysis of Hydrolyzable Tannins and Other Phenolic Compounds in Emblic Leafflower (Phyllanthus emblica L.) Fruits by High Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry

Phenolic compounds were extracted from dried emblic leafflower (Phyllanthus emblica L.) fruits with methanol and separated by Sephadex LH-20 column chromatography. The raw extracts and fractions were analyzed with HPLC coupled with diode array UV spectroscopy, electrospray ionization mass spectrometry, and tandem mass spectrometry. Mucic acid gallate, mucic acid lactone gallate, monogalloylglucose, gallic acid, digalloylglucose, putranjivain A, galloyl-HHDP-glucose, elaeocarpusin, and chebulagic acid were suggested to be the most abundant compounds in the crude methanol extracts of the fruits. In addition, 144 peaks were detected, of which 67 were tentatively identified mostly as ellagitannins, flavonoids, and simple gallic acid derivatives in the fractions. The results indicated the presence of neochebulagic acid, isomers of neochebuloyl galloylglucose, chebuloyl neochebuloyl galloylglucose, ellagic acid glycosides, quercetin glycosides, and eriodictyol coumaroyl glycosides in the fruits. The study provides a systematic report of the retention data and characteristics of UV, MS, and MS/MS spectra of the phenolic compounds in the fruits of emblic leafflower. The fruits of two varieties (Ping Dan No 1 and Fruity) from Guangxi Province differed from those of wild Tian Chuan emblic leafflower from Fujian Province in the content and profile of phenolic compounds.     

Polyphenols of pseudostem of different banana cultivars and their antioxidant activities

The present investigation is focused on potential use of banana pseudostem (BPS), which otherwise is disposed off as a waste or incinerated, as a source of polyphenols or antioxidants. The total phenolics (TP) and total flavonoids (TF) in various solvent extracts of pseudostem (PS) of different banana cultivars varied from 7.58 to 291 mg gallic acid equivalent (GAE/g of extract) and from 4 to 80 mg catechin equivalent (CE/g of extract), respectively. Acetone extract showed high antioxidant activity (AOA) in all of the in vitro models tested, whereas methanol extract exhibited high metal chelating activity. Among the banana cultivars, Nanjanagudu Rasabale (NR) showed the highest TP (291 mg GAE/g of extract), TF (80 mg CE/g of extract), and AOA. A detailed study on phenolic acids by reverse phase HPLC and ESI-MS revealed the presence of phenolic acids such as gentisic acid, (+)-catechin, protocatechuic acid, caffeic acid, ferulic acid, and cinnamic acid in cultivar NR. The differences in AOA of banana cultivars are in accordance with their phenolic and flavonoid concentrations.