Identification of natural oak lactone precursors in extracts of american and French oak woods by liquid chromatography-tandem mass spectrometry

A method for the screening of potential natural oak lactone precursors in oak wood extracts using LC-MS/MS combined with information-dependent acquisition was developed. The method was applied to extracts of American and French oak woods. As a result, cis-3-methyl-4-galloyloxyoctanoic acid (ring-opened cis-oak lactone gallate), (3S,4S)- and (3S,4R)-3-methyl-4-O-beta-D-glucopyranosyloctanoic acid (ring-opened cis- and trans-oak lactone glucoside), and (3S,4S)-3-methyl-4-O-(6'-O-galloyl)-beta-D-glucopyranosyloctanoic acid (ring-opened cis-oak lactone galloylglucoside) were identified as natural oak lactone precursors in the extracts by comparison with the respective synthetic reference compounds. In addition, the ring-opened oak lactone rutinoside was tentatively identified in the extracts. Three apparent isomers of the ring-opened cis-oak lactone galloylglucoside were also observed.     

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.     

Volatile compounds and sensorial characterization of wines from four Spanish denominations of origin, aged in Spanish Rebollo (Quercus pyrenaica Willd.) oak wood barrels

The evolution of almost 40 oak-related volatile compounds and the sensorial characteristics of red wines from four Spanish denominations of origin (DOs) (Bierzo, Toro, Ribera de Duero, and Rioja) during aging in barrels made of Rebollo oak wood, Quercus pyrenaica, were studied and compared to the same wines aged in American and French oak barrels. Each oak wood added unique and special characteristics to the wine, and in addition, each wine showed a different ability to extract the compounds, which result in these characteristics from the oak wood. In general, wines aged in Q. pyrenaica wood were characterized by high levels of eugenol, guaiacol, and other volatile phenols. In regards to compounds like cis-whiskylactone or maltol, the behavior of this wood is very similar to that of American oaks. When considering phenolic aldehydes and ketones, the levels of these compounds are intermediate between those of French and American woods and depend greatly on the type of wine. The type of oak, on the other hand, does not affect the chromatic characteristics of the wines. In sensory analysis, the biggest differences are found in the olfactory phase. Among the four DOs studied, wine aged in Q. pyrenaica presented the highest notes of wood, with more aromas of roasting, toasting, milky coffee, spices, or wine-wood interactions. The wines aged in barrels made of Q. pyrenaica wood were highly regarded, and preference was shown for them over those same wines when they had been aged in barrels of American or French oak.     

Characteristic volatiles from young and aged fruiting bodies of wild Polyporus sulfureus (Bull.:Fr.) Fr

The volatile compounds of fresh fruiting bodies of wild Polyporus sulfureus (Bull.:Fr.) Fr. growing on oak trees were isolated by continuous liquid-liquid extraction (CLLE) and investigated by high-resolution gas chromatography-mass spectrometry (HRGC-MS) on two GC columns of different polarity (DB-5 and ZB-WAX), and by gas chromatography-olfactometry (GC-O). A total of 40 major volatile compounds from the young samples were identified and semiquantified. Five odorous compounds were determined to be responsible for the characteristic flavor of young Polyporus sulfureus: 1-octen-3-one, 1-octen-3-ol, 3-methylbutanoic acid, phenylethanol, and phenylacetic acid. Four volatiles investigated by GC-O and detected by GC-MS were determined as the characteristic odorants of aged species: 2-methylpropanoic acid, butanoic acid, 3-methylbutanoic acid, and phenylacetic acid. The comparative results revealed that the volatile composition of the fruiting bodies even from the same fungal species may greatly vary with its host, location, and age.     

C(13)-Norisoprenoid glucoconjugates from lulo (Solanum quitoense L.) leaves

With the aid of multilayer coil countercurrent chromatography, subsequent acetylation, and liquid chromatographic purification of a glycosidic mixture obtained from lulo (Solanum quitoense L.) leaves, three C(13)-norisoprenoid glucoconjugates were isolated in pure form. Their structures were elucidated by NMR, MS, and CD analyses to be the novel (6R,9R)-13-hydroxy-3-oxo-alpha-ionol 9-O-beta-D-glucopyranoside (4a), the uncommon (3S,5R,8R)-3, 5-dihydroxy-6,7-megastigmadien-9-one 5-O-beta-D-glucopyranoside (citroside A) (5a), and the known (6S,9R)-vomifoliol 9-O-beta-D-glucopyranoside (6a). Enzymatic treatment of compound 5a showed the formation of 3-hydroxy-7,8-didehydro-beta-ionone (7), an important lulo peeling volatile, which in its turn after chemical reduction and heated acid catalyzed rearrangement generates beta-damascenone (9) and 3-hydroxy-beta-damascone (10).     

Enantioselective analysis of methyl-branched alcohols and acids in rhubarb (Rheum rhabarbarum L.) stalks

The enantiomeric compositions of 2-methylbutanol (1), 4-methylhexanol (2), 2-methylbutanoic acid (3), and 4-methylhexanoic acid (4) present in rhubarb (Rheum rhabarbarum L.) stalks were determined. Enantiodifferentiation was achieved via multidimensional gas chromatography using heptakis(2,3,6-tri-O-ethyl)-beta-cyclodextrin as a chiral stationary phase. For all compounds the enantiomeric ratios were in favor of the (R)-enantiomers. The alcohols (1 and 2) exhibited generally high excesses of the (R)-enantiomers, the ratios varying slightly from batch to batch. For the acid (3) a rather narrow range averaging 65% (R):35% (S) was observed. The procedure applied to isolate the volatiles (vacuum headspace technique, simultaneous distillation--extraction, liquid--liquid extraction) had no significant impact on the enantiomeric ratios. The study describes for the first time a plant used as food material in which 2-methyl-branched volatiles are not nearly exclusively present as (S)-enantiomers. This information enlarges the current regulatory knowledge regarding the classification of these important flavor compounds as "natural" on the basis of their enantiomeric ratios.     

Spectroscopic identification and antioxidant activity of glucosylated carotenoid metabolites from Cydonia vulgaris fruits

Carotenoid metabolites are common plant constituents with significant importance for the flavor and aroma of fruits. Three new carotenoid derivatives, (2E,4E)-8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 1-O-beta-D-glucopyranosyl ester (1), (2Z,4E)-8-beta-D-glucopyranosyloxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid (3), and 3,9-dihydroxymegastigmast-5-ene-3-O-[beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (5), as well as three known compounds, have been isolated from the ethanolic extract of peels of Cydonia vulgaris, the fruit of a shrub belonging to the same family as the apple. All the compounds were identified by spectroscopic techniques, especially 1D and 2D NMR. Antioxidant activities of all the isolated metabolites were assessed by measuring their ability to scavenge DPPH radical and superoxide radical (O2*-) and to induce the reduction of Mo(VI).     

Abscisic acid related compounds and lignans in prunes (Prunus domestica L.) and their oxygen radical absorbance capacity (ORAC)

Four new abscisic acid related compounds (1-4), together with (+)-abscisic acid (5), (+)-beta-D-glucopyranosyl abscisate (6), (6S,9R)-roseoside (7), and two lignan glucosides ((+)-pinoresinol mono-beta-D-glucopyranoside (8) and 3-(beta-D-glucopyranosyloxymethyl)-2- (4-hydroxy-3-methoxyphenyl)-5-(3-hydroxypropyl)-7-methoxy-(2R,3S)-dihydrobenzofuran (9)) were isolated from the antioxidative ethanol extract of prunes (Prunus domestica L.). The structures of 1-4 were elucidated on the basis of NMR and MS spectrometric data to be rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (1), rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid 3'-O-beta-d-glucopyranoside (2), rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (3), and rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxabicyclo[3,2,1]- oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (4). The antioxidant activities of these isolated compounds were evaluated on the basis of oxygen radical absorbance capacity (ORAC). The ORAC values of abscisic acid related compounds (1-7) were very low. Two lignans (8 and 9) were more effective antioxidants whose ORAC values were 1.09 and 2.33 micromol of Trolox equiv/micromol, respectively.     

Sensory-directed identification of taste-active ellagitannins in American (Quercus alba L.) and European oak wood (Quercus robur L.) and quantitative analysis in bourbon whiskey and oak-matured red wines

Aimed at increasing our knowledge on the sensory-active nonvolatiles migrating from oak wood into alcoholic beverages upon cooperaging, an aqueous ethanolic extract prepared from oak wood chips (Quercus alba L.) was screened for its key taste compounds by application of the taste dilution analysis. Purification of the compounds perceived with the highest sensory impacts, followed by liquid chromatography/mass spectrometry as well as one-dimensional and two-dimensional NMR experiments, revealed the ellagitannins vescalagin, castalagin, and grandinin, the roburins A-E, and 33-deoxy-33-carboxyvescalagin as the key molecules imparting an astringent oral sensation. To the best of our knowledge, 33-deoxy-33-carboxyvescalagin has as yet not been reported as a phytochemical in Q. alba L. In addition, the sensory activity of these ellagitannins was determined for the first time on the basis of their human threshold concentrations and dose/response functions. Furthermore, the ellagitannins have been quantitatively determined in extracts prepared from Q. alba L. and Quercus robur L., respectively, as well as in bourbon whiskey and oak-matured red wines, and the sensory contribution of the individual compounds has been evaluated for the first time on the basis of dose/activity considerations.     

Investigation of the tuber constituents of maca (Lepidium meyenii Walp.)

Lepidium meyenii, known in South America as maca, has received attention worldwide as a powerful energizer that improves physical and mental conditions and increases fertility. Because of these reports, we investigated the secondary metabolites of the tuber of maca. The methanol extract of the tuber of maca contained, in addition to free sugars and amino acids, the following: uridine, malic acid and its benzoyl derivative, and the glucosinolates, glucotropaeolin and m-methoxyglucotropaeolin. Because glucosinolates and their derived products have received increasing attention due to their biological activities, the occurrence of glucosinolate degradation products in the hexane extract was also investigated, and benzylisothiocyanate and its m-methoxy derivative were isolated. The two glucosinolates were semiquantified by HPLC, and benzylisothiocyanate was semiquantified by GC/MS. The methanol extract of maca tuber also contained (1R,3S)-1-methyltetrahydro-beta-carboline-3-carboxylic acid, a molecule which is reported to exert many activities on the central nervous system.     

Volatile compounds in a spanish red wine aged in barrels made of Spanish, French, and American oak wood

A red Rioja wine was aged in barrels made of Spanish oak wood (Quercus robur, Quercus petraea,Quercus pyrenaica, and Quercus faginea) during 21 months. The concentrations of some volatile compounds [syringaldehyde, vanillin, eugenol, maltol, guaiacol, 4-ethylphenol, cis and trans isomers of beta-methyl-gamma-octalactone, 2-furfuraldehyde, 5-methyl-2-furfuraldehyde, 5-(hydroxymethyl)-2-furfuraldehyde, and furfuryl alcohol] were studied in these wines and compared with those of the same wine aged in barrels made from French oak of Q. robur (Limousin, France) and Q. petraea (Allier, France) and American oak of Quercus alba (Missouri). Similar concentrations of these compounds were found in wines aged in Spanish and French oak wood barrels, and significantly different concentrations were found with respect to wines aged in barrels made of American oak wood, indicating a different behavior. Thus, wines with different characteristics were obtained, depending on the kind of wood. Also, the kind of wood had an important influence on sensory characteristics of wine during the aging process. Spanish oak wood from Q. robur, Q. petraea, and Q. pyrenaica can be considered to be suitable for barrel production for quality wines, because a wine aged in barrels made of these Spanish oak woods showed similar and intermediate characteristics to those of the same wine aged in French and American oak woods usually used in cooperage.     

Identifying new volatile compounds in toasted oak

Toasting wood to be used in barrels for aging wine produces a great number of volatile and odiferous compounds. Three new volatile odorous compounds in toasted oak were identified. Analysis by high-performance gas chromatography of toasted oak extracts, combined with olfactory detection, enabled various chromatographic peaks with these specific aromas to be isolated. These same odors were simultaneously studied by heating glucose both with and without proline and phenylalanine. Aromatic compounds of interest were identified thanks to a combination of gas chromatography and both mass and infrared spectrometry. An analysis RMN was also used. Hydroxymaltol, 2,5-furanedicarbaldehyde, and furylhydroxymethyl ketone have been detected in extract of toasted oak wood. These molecules may be formed by direct pyrolysis of sugar or Maillard reactions. The acetylformoine was not detected in extract of toasted oak wood, whereas it was detected in heated extracts of various sugars and sugars mixtures with amino acids.     

Characterization of volatiles in extracts from oak chips obtained by accelerated solvent extraction (ASE)

Sixteen commercially available oak chips, differing in origin (French or American) and toasting level, were extracted by an accelerated solvent extraction method and characterized by their volatile composition. About 80 compounds were identified and quantified, a great part of them from the thermodegradation of lignin and cellulose. One furanone (solerone) and two C-13 norisoprenoids (3-oxo-alpha-ionol and a 3-oxoretro-alpha-ionol isomer) were also tentatively identified and reported for the first time in toasted wood. Quantitative data demonstrated the oak chips to be not so different from the composition of light- or medium-toasted wood barrels, which was reported by other authors. The same data suggest that toasting level had the strongest influence on the volatile composition of chip samples. Phenyl ketones, volatile phenols, and some furanic compounds were the most influenced. On the other hand, the influence of wood origin was found to be weaker.     

Solanidine hydrolytic extraction and separation from the potato (Solanum tuberosum L.) vines by using solid-liquid-liquid systems

Solanidine is a steroidal aglycon of potato (Solanum tuberosum L.) glycoalkaloids and a very important precursor for the synthesis of hormones and some pharmacologically active compounds. Glycoalkaloids are hydrolyzed by mineral acid, yielding solanidine. This paper deals with the kinetics of solanidine hydrolytic extraction in different solid-liquid-liquid systems. The dried and milled potato (S. tuberosum L.) vines were used as a source of glycoalkaloids and as the solid phase. The solutions of hydrochloric acid in 2 and 10% (w/v) aqueous acetic acid, in 50% (volume) aqueous methanol, and in 50% (volume) aqueous ethanol were first liquid phase, and the medium for glycoalkaloid extraction from potato vines and their hydrolysis to solanidine. The chloroform, trichloroethylene, or carbon tetrachloride were the second, organic, liquid phase and the medium for solanidine extraction. This procedure combines three different processes: extraction of glycoalkaloids from potato vines, their hydrolysis to solanidine, and the extraction of solanidine, in a single step. The term hydrolytic extraction of solanidine was used for these processes. The purpose of the paper was to choose an optimal solid-liquid-liquid system for solanidine extraction and to define the procedure for its isolation from the organic liquid phase. The best degree of solanidine hydrolytic extraction (DHE) of more than 98% was achieved when 10% (w/v) hydrochloric acid in 50% (volume) methanol were the first liquid phase and chloroform was the second liquid phase, after 90 min. The yield of solanidine (q(S)) under these conditions is calculated to be 0.24 g/100 g of potato vines. Approximately 78% of the maximal possible yield of solanidine was isolated from chlorofom liquid phase. The IR and MS spectra of isolated solanidine were recorded.     

Properties of diphenolase from Vanilla planifolia (Andr.) shoot primordia cultured in vitro.

Properties of diphenolase (PPO, EC1.10.3.1) from vanilla (Vanilla planifolia Andr.) shoot primordia culture were investigated. Two pH optima of the enzyme extraction at pH 6 and 8 were found. Nevertheless, the enzymes shared the same optimum pH of activity-between pH 3 and 4. Sodium dodecyl sulfate slightly improved diphenolase extraction but caused a 3-fold increase in its specific activity. The extracts of pH 6 and 8.0 revealed three isozyme bands after polyacrylamide gel electrophoresis-two of them were similar in both extracts and two distinct. The enzyme showed high thermal stability-no loss was observed after 120 min at 50 degrees C. Diethyldithiocarbamic acid, ethylenediaminetetracetic acid disodium salt, ethylene glycol bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, L-ascorbic acid, dithiothreitol, glutathione (reduced), and beta-mercaptoethanol were found to be potent inhibitors of the diphenolase studied. The enzyme showed also monophenolase activity. Km and Vmax were calculated with monophenols [p-coumaric acid, 3-(p-hydroxyphenyl)propionic acid, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid] and with diphenols (caffeic acid, hydrocaffeic acid, chlorogenic acid, 4-methylcatechol, protocatechuic aldehyde and acid, and 3,4-dihydroxyphenylalanine). The highest Vmax was found with 4-hydroxybenzyl alcohol and the greatest affinity to protocatechuic acid, respectively-the most abundant monophenol and one of the least abundant o-diphenols in the studied Vanilla tissue.     

Determination of amino acids from the leaves of evergreen oak (Quercus ilex L.) at four different stages of decomposition

Summary It was established by weak ion exchange resin chromatography that 17 amino acids in proteins and 15 free amino acids account for almost all of the amino acids present in the leaves of evergreen oak (Quercus ilex L.) in four different stages of decomposition. No significant change in the amounts of these amino acids was observed in the course of decomposition. The amino acid content of evergreen oak leaves is comparable to that in pine needles and beech leaves.     

Isolation, structure determination, and sensory activity of mouth-drying and astringent nitrogen-containing phytochemicals isolated from red currants (Ribes rubrum)

Application of chromatographic separation and taste dilution analyses recently revealed, besides a series of flavon-3-ol glycosides and (E)/(Z)-aconitic acid, four nitrogen-containing phytochemicals as the key astringent and mouth-drying compounds in red currants (Ribes rubrum). The isolation and structure determination of the astringent indoles 3-carboxymethyl-indole-1-N-beta-D-glucopyranoside (1) and 3-methylcarboxymethyl-indole-1-N-beta-D-glucopyranoside (2), as well as the astringent, noncyanogenic nitriles 2-(4-hydroxybenzoyloxymethyl)-4-beta-D-glucopyranosyloxy-2(E)-butenenitrile (3) and 2-(4-hydroxy-3-methoxybenzoyloxymethyl)-4-beta-D-glucopyranosyloxy-2(E)-butenenitrile (4) by means of 1D/2D NMR, LC-MS/MS, and UV-vis spectroscopy are reported. The structures of compounds 1 and 2 were confirmed by synthesis. Using the recently developed half-tongue test, human recognition thresholds for the astringent and mouth-drying nitrogen compounds were determined to be between 0.0003 and 5.9 micromol/L (water). In particular, the extraordinarily low threshold of 0.0003 micromol/L evaluated for the indole 1 represents the lowest recognition threshold of any astringent phytochemical reported to date.     

Sensory-guided decomposition of red currant juice (Ribes rubrum) and structure determination of key astringent compounds

Sequential application of solvent extraction, gel permeation chromatography, and RP-HPLC in combination with taste dilution analyses, followed by LC-MS and 1D/2D NMR experiments, led to the discovery and structure determination of 25 key astringent compounds of red currant juice. Besides several flavonol glycosides, in particular, 3-carboxymethyl-indole-1-N-beta-D-glucopyranoside, 3-methylcarboxymethyl-indole-1-N-beta-D-glucopyranoside, and a family of previously not identified compounds, namely, 2-(4-hydroxybenzoyloxymethyl)-4-beta-D-glucopyranosyloxy-2(E)-butenenitrile, 2-(4-hydroxy-3-methoxybenzoyloxymethyl)-4-beta-D-glucopyranosyloxy-2(E)-butenenitrile, (E)-6-[3-hydroxy-4-(O-beta-D-glucopyranosyl)phenyl]-5-hexen-2-one named dehydrorubrumin, and (3E,5E)-6-[3-hydroxy-4-(O-beta-D-glucopyranosyl)phenyl]-3,5-hexadien-2-one named rubrumin, have been identified. Determination of the oral astringency thresholds by means of the half-tongue test revealed that the lowest thresholds of 0.3 and 1.0 nmol/L were found for the nitrogen-containing 3-carboxymethyl-indole-1-N-beta-D-glucopyranoside and 3-methylcarboxymethyl-indole-1-N-beta-D-glucopyranoside, which do not belong to the group of plant polyphenols.     

LDL-antioxidant pterocarpans from roots of Glycine max (L.) Merr

The methanolic root extract of Glycine max (L.) Merr. was chromatographed, which yielded 10 flavonoids, including three isoflavones 1-3, five pterocarpans 4-8, one flavonol 9, and one anthocyanidin 10. All isolated compounds were examined for LDL-antioxidant activities using four different assay systems on the basis of Cu2+-mediated oxidation. Among them, seven compounds showed potent LDL-antioxidant activities in the thiobarbituric acid reactive substances (TBARS) assay, the lag time of conjugated diene formation, relative electrophoretic mobility (REM), and fragmentation of apoB-100 on copper-mediated LDL oxidation. Three pterocarpans 4, 6, and 7, never reported as LDL-antioxidant, showed potent activities with IC50 values of 19.8, 0.9, 45.0 microM, respectively, in comparison with probucol (IC50 = 5.6 microM) as positive control. Interestingly, coumestrol 6 (IC50 = 0.9 microM) showed 20 times more activity in the TBARS assay than genistein (IC50 = 30.1 microM) and daidzein (IC50 = 21.6 microM), representative antioxidants in soybean. Moreover, coumestrol 6 had an extended lag time of 190 min at 3.0 microM in measuring conjugated diene formation, while both genistein (120 min) and daidzein (93 min) lag times were extended to less than 120 min at the same concentration.     

Antioxidant constituents in the fruits of Luffa cylindrica (L.) Roem

Hydrophilic antioxidant constituents in the fruits of the vegetable Luffa cylindrica (L.) Roem (sponge gourds) were separated by an antioxidant-guided assay to yield eight compounds: p-coumaric acid (1), 1-O-feruloyl-beta-D-glucose (2), 1-O-p-coumaroyl-beta-D-glucose (3), 1-O-caffeoyl-beta-D-glucose (4), 1-O-(4-hydroxybenzoyl)glucose (5), diosmetin-7-O-beta-D-glucuronide methyl ester (6), apigenin-7-O-beta-D-glucuronide methyl ester (7), and luteolin-7-O-beta-D-glucuronide methyl ester (8). The eight compounds were isolated by high-speed countercurrent chromatography and identified by electrospray ionization-mass spectrometry and NMR analysis, and the antioxidant activity was evaluated by the radical scavenging effect on the 1,1-diphenyl-2-picrylhydrazyl radical. High-performance liquid chromatography analysis showed that a total amount of the eight compounds in the dried gourds without skin was about 1%. The results demonstrate that the consumption of sponge gourds can supply some antioxidant constituents to human body.