Content of phenolic acids and ferulic acid dehydrodimers in 17 rye (Secale cereale L.) varieties

The contents of pnenolic acids and ferulic acid dehydrodimers were quantified by HPLC analysis after alkaline hydrolysis in kernels of 17 rye (Secale cereale L.) varieties grown in one location in Denmark during 1997 and 1998. Significant variations (P < 0.05) with regard to the concentration of the analyzed components were observed among the different rye varieties and also between different harvest years. However, the content of phenolic acids in the analyzed rye varieties was narrow compared to cereals such as wheat and barley. The concentration of ferulic acid, the most abundant phenolic acid ranged from 900 to 1170 microgram g(-1) dry matter. The content in sinapic acid ranged from 70 to 140 microgram g(-1) dry matter, p-coumaric acid ranged from 40 to 70 microgram g(-1) dry matter, and caffeic, p-hydroxybenzoic, protocatechuic, and vanillic acids were all detected in concentrations less than 20 microgram g(-1) dry matter. The most abundant ferulic acid dehydrodimer 8-O-4 -DiFA was quantified in concentrations from 130 to 200 microgram g(-1) dry matter followed by 8,5 -DiFA benzofuran form (50-100 microgram g(-1) dry matter), 5,5 -DiFA (40-70 microgram g(-1) dry matter), and 8,5 -DiFA (20-40 microgram g(-1) dry matter).     

Antioxidant effects of phenolic rye (Secale cereale L.) extracts, monomeric hydroxycinnamates, and ferulic acid dehydrodimers on human low-density lipoproteins

Dietary antioxidants that protect low-density lipoprotein (LDL) from oxidation may help to prevent atherosclerosis and coronary heart disease. The antioxidant activities of purified monomeric and dimeric hydroxycinnamates and of phenolic extracts from rye (whole grain, bran, and flour) were investigated using an in vitro copper-catalyzed human LDL oxidation assay. The most abundant ferulic acid dehydrodimer (diFA) found in rye, 8-O-4-diFA, was a slightly better antioxidant than ferulic acid and p-coumaric acid. The antioxidant activity of the 8-5-diFA was comparable to that of ferulic acid, but neither 5-5-diFA nor 8-5-benzofuran-diFA inhibited LDL oxidation when added at 10-40 microM. The antioxidant activity of the monomeric hydroxycinnamates decreased in the following order: caffeic acid > sinapic acid > ferulic acid > p-coumaric acid. The antioxidant activity of rye extracts was significantly correlated with their total content of monomeric and dimeric hydroxycinnamates, and the rye bran extract was the most potent. The data suggest that especially rye bran provides a source of dietary phenolic antioxidants that may have potential health effects.     

Hydroxycinnamic acids as affected by different fertilization of Rebula grapevines

The phenolic composition of white grapes is important since browning reactions may dramatically compromise the overall quality of musts and wines. Only few scientific contributions are available, which investigated how fertilization can influence this class of compounds. Thus, the aim of this work was to examine the effects of NPK soil fertilization coupled with soil or foliar applications of Mg, Fe, and Zn on the concentrations of K, Mg, Fe, and Zn of grape berries and leaf petioles of potted Rebula grapevines (Vitis vinifera L.), also revealing the change of hydroxycinnamic acids in grape juice. The results obtained over the three‐year study (2009–2011) showed that NPK fertilization positively affected K and Zn concentrations of petioles and grape berries, and negatively Mg. In addition, K (synergistic) and Mg (antagonistic) had an influence on Zn uptake. Our findings suggest that the supply of NPK was profitable for a reduction of trans‐caftaric acid in grape juice, while only few differences were observed with further application of nutrients.     

Oxidation of ferulic acid or arabinose-esterified ferulic acid by wheat germ peroxidase

The oxidation of ferulic acid (FA) or 5-O-(trans-feruloyl)-L-arabinose (EFA) by a purified wheat germ peroxidase was followed by UV spectrophotometry and high-performance liquid chromatography using an electrochemical detection. Wheat peroxidase (POD) exhibits a ping-pong bireactant mechanism forming phenoxy radicals more rapidly from FA than from EFA in routine assay conditions. When both the free and the esterified forms of FA are present, the reverse was found. This result could be due to a nonenzymatic cooxidation of FA by the phenoxy radicals of EFA leading to the formation of phenoxy radicals of FA and the EFA regeneration. Addition of ascorbic acid (AA) provokes a delay of FA consumption. AA reduced very rapidly the phenoxy radicals formed by POD back to initial phenol avoiding the formation of ferulate dimers until it was completely oxidized in dehydroascorbic acid. Conversely, cysteine addition slowed but did not delay the FA consumption. The thiol reduced a fraction of the phenoxy radicals produced by wheat POD and was oxidized into cystine, while the other part of phenoxy radicals formed ferulate dimers. These results could be of interest to understand the POD effect on the wheat dough rheological properties.     

Hydroxycinnamic acid ethyl esters as precursors to ethylphenols in wine

A method for determining ethyl coumarate and ethyl ferulate in wine using GC-MS with deuterium-labeled analogues has been developed and used to measure the evolution of these two esters during the production of two commercial monovarietal red wines, cv. Grenache and Shiraz. During fermentation, the concentration of ethyl coumarate rose from low levels to 0.4 mg/L in Grenache and 1.6 mg/L in Shiraz wines. These concentrations then increased further during barrel aging to 1.4 and 3.6 mg/L, respectively. The concentration of ethyl ferulate was much lower, reaching a maximum of only 0.09 mg/L. Conversion of ethyl coumarate and ethyl ferulate to their corresponding ethylphenols was observed during fermentations of a synthetic medium with two strains of Dekkera bruxellensis (AWRI 1499 and AWRI 1608), while a third (strain AWRI 1613) produced no ethylphenols at all from these precursors. Strains AWRI 1499 and 1608 produced 4-ethylphenol from ethyl coumarate in 68% and 57% yields, respectively. The corresponding yields of 4-ethylguaiacol from ethyl ferulate were much lower, 7% and 3%. Monitoring of ethyl coumarate and ethyl ferulate concentration during the Dekkera fermentations showed that the selectivity for ethylphenol production according to yeast strain and the precursor was principally a result of variation in esterase activity. Consequently, ethyl coumarate can be considered to be a significant precursor to 4-ethylphenol in wines affected by these two strains of Brettanomyces/Dekkera yeast, while ethyl ferulate is not an important precursor to 4-ethylguaiacol.     

Release of ferulic acid from oat hulls by Aspergillus ferulic acid esterase and trichoderma xylanase

Oat hulls, an agricultural byproduct, contain a relatively high amount of ferulic acid (FA; 4-hydroxy-3-methoxycinnamic acid), which is believed to be inhibitory to oat hull biodegradability by rumen microorganisms. In this paper, Aspergillus ferulic acid esterase (FAE) was investigated for its ability to release FA from oat hulls. The objectives were to determine the effects of particle size of oat hulls (ground to pass through 1 mm and 250 microm screens and a 100 microm sieve) on release of FA by FAE both in the presence and in the absence of Trichoderma xylanase. The results show that the release of FA by FAE was dependent upon the particle size of oat hulls (< or = 250 microm). In the absence of Trichoderma xylanase, little FA was released by FAE. In the presence of Trichoderma xylanase, there was a significant release of FA by FAE, indicating a synergistic interaction between FAE and Trichoderma xylanase on release of FA from oat hulls. These results indicate that FAE is able to break the ester linkage between FA and the attached sugar, releasing FA from oat hulls. This may leave the remainder of the polysaccharides open for further hydrolytic attack by rumen microorganisms. It is likely that removing FA from oat hulls could improve rumen biodegradability, thus improving the nutritional value of oat hulls.     

Peroxidase-catalyzed oligomerization of ferulic acid esters

Valuable information about possible types of linkages, reaction mechanisms, and sequences for oxidative coupling of phenolic compounds in planta is available from in vitro model systems. Ferulate oligomers were generated in a system using ethyl ferulate, peroxidase, and hydrogen peroxide under various conditions. A molar ferulate/H2O2 ratio of 1:1, an ethanol level of 30% in an aqueous sodium phosphate buffer (pH 6.0), and a reaction time of 10 min were considered to be ideal to produce maximal proportions of ferulate trimers and tetramers from ethyl ferulate as starting material. The dominant trimer and tetramer were each isolated from the reaction mixture and identified as 8-O-4/8-5(cyclic)-dehydrotriferulic acid triethyl ester and 8-5(cyclic)/4-O-5/8-5(cyclic)-dehydrotetraferulic acid tetraethyl ester. The structure of the 8-O-4/8-5(cyclic)-dehydrotriferulic acid triethyl ester revealed that a third ferulate unit is bound to a preformed 8-O-4-diferulate dimer, a surprising reaction sequence considering the dominance of 8-5-coupled dimers among dehydrodiferulates in H2O2/peroxidase-based model reactions. As 4-O-5-coupling is not favored in the dimerization process of ferulates, the main tetramer isolated in this study is probably formed by 4-O-5-coupling of two preformed 8-5(cyclic)-diferulates, a logical step in analogy with reactions occurring in lignin biosynthesis.     

Sulfated ferulic acid is the main in vivo metabolite found after short-term ingestion of free ferulic acid in rats

The bioavailability of ferulic acid (FA; 3-methoxy-4-hydroxycinnamic acid) and its metabolites was investigated in rat plasma and urine after an oral short-term ingestion of 5.15 mg/kg of FA. Free FA, glucuronoconjugates, and sulfoconjugates were quickly detected in plasma with a peak of concentration found 30 min after ingestion. Sulfoconjugates were the main derivates ( approximately 50%). In urine, the cumulative excretion of total metabolites reached a plateau 1.5 h after ingestion, and approximately 40% were excreted by this way. Free FA recovered in urine represented only 4.9 +/-1.5% of the native FA consumed by rats. Glucuronoconjugates and sulfoconjugates represented 0.5 +/- 0.3 and 32.7 +/- 7.3%, respectively. These results suggested that a part of FA incorporated in the diet was quickly absorbed and largely metabolized in sulfoconjugates before excretion in urine.     

Sinapate dehydrodimers and sinapate-ferulate heterodimers in cereal dietary fiber

Two 8-8-coupled sinapic acid dehydrodimers and at least three sinapate-ferulate heterodimers have been identified as saponification products from different insoluble and soluble cereal grain dietary fibers. The two 8-8-disinapates were authenticated by comparison of their GC retention times and mass spectra with authentic dehydrodimers synthesized from methyl or ethyl sinapate using two different single-electron metal oxidant systems. The highest amounts (481 microg/g) were found in wild rice insoluble dietary fiber. Model reactions showed that it is unlikely that the dehydrodisinapates detected are artifacts formed from free sinapic acid during the saponification procedure. The dehydrodisinapates presumably derive from radical coupling of sinapate-polymer esters in the cell wall; the radical coupling origin is further confirmed by finding 8-8 and 8-5 (and possibly 8-O-4) sinapate-ferulate cross-products. Sinapates therefore appear to have an analogous role to ferulates in cross-linking polysaccharides in cereal grains and presumably grass cell walls in general.     

Antioxidant properties of ferulic acid and its related compounds

Antioxidant activity of 24 ferulic acid related compounds together with 6 gallic acid related compounds was evaluated using several different physical systems as well as their radical scavenging activity. The radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) decreased in the order caffeic acid > sinapic acid > ferulic acid > ferulic acid esters > p-coumaric acid. In bulk methyl linoleate, test hydroxycinnamic acids and ferulic acid esters showed antioxidant activity in parallel with their radical scavenging activity. In an ethanol-buffer solution of linoleic acid, the activity of test compounds was not always associated with their radical scavenging activity. Ferulic acid was most effective among the tested phenolic acids. Esterification of ferulic acid resulted in increasing activity. The activity of alkyl ferulates was somewhat influenced by the chain length of alcohol moiety. When the inhibitory effects of alkyl ferulates against oxidation of liposome induced by AAPH were tested, hexyl, octyl, and 2-ethyl-1-hexyl ferulates were more active than the other alkyl ferulates. Furthermore, lauryl gallate is most effective among the tested alkyl gallates. These results indicated that not only the radical scavenging activity of antioxidants, but also their affinity with lipid substrates, might be important factors in their activity.     

阿魏酸与肉桂酸对杉木化感作用的生物评价

通过绿豆、大豆、黄瓜、空心菜及玉米等种子发芽的生物评价,分析研究阿魏酸和肉桂酸对杉木化感作用结果表明,阿魏酸对5种蔬菜种子作用表现为高浓度抑制萌发和生长,随浓度的降低其抑制作用逐渐减弱。肉桂酸除对空心菜发芽表现为高浓度促进生长外,对其余种子作用表现为高浓度抑制萌发和生长,随浓度的降低其抑制作用逐渐减弱。     

Reactivity of ferulic acid and its derivatives toward hydrogen peroxide and peracetic acid.

The reactions of ferulic acid and its derivatives with hydrogen peroxide and peracetic acid in lignin-retaining bleaching conditions have been investigated to determine their susceptibility to oxidative degradation. The conjugated side chain of ferulic acid and its etherified or esterified derivative was shown to be fairly stable, especially to hydrogen peroxide. The major reaction was trans-cis isomerization that possibly involved a radical mechanism but did not cause bond cleavage. The peracetic acid reaction increased the rate of trans-cis isomerization and was also accompanied by a minor cleavage of the side chain. Esterification did not have a substantial effect on the reactivity of ferulic acid, but 4-O-etherification significantly stabilized it against these two oxidants. By contrast, aldehyde substitution tremendously enhanced the susceptibility of the cinnamyl side chain to oxidative degradation, as evidenced by an intensive degradation of coniferaldehyde.     

Regioselective dimerization of ferulic acid in a micellar solution.

Dehydrodimers of hydroxycinnamates play an important role in the cross-linking of plant cell walls. An aqueous solution of quaternary ammonium salts with a long aliphatic chain is known to spontaneously organize itself into micelles with the ionic part at the outer sphere. It is shown that regioisomeric ferulic acid dehydrodimers can be obtained in one step from trans-ferulic acid after attachment to these micelles and using the biomimetic peroxidase-H2O2 system. The surfactant hexadecyltrimethylammonium hydroxide yielded trans-4-(4-hydroxy-3-methoxybenzylidene)-2-(4-hydroxy-3-methoxyphenyl)-5-oxotetrahydrofuran-3-carboxylic acid (25%), (E,E)-4,4'-dihydroxy-5,5'-dimethoxy-3,3'-bicinnamic acid (21%), and trans-5-[(E)-2-carboxyvinyl]-2-(4-hydroxy-3-methoxyphenyl)-7-methoxy-2,3-dihydrobenzofuran-3-carboxylic acid (14%), whereas the surfactant tetradecyltrimethylammonium bromide gave 4-cis, 8-cis-bis(4-hydroxy-3-methoxyphenyl)-3,7-dioxabicyclo[3.3.0]octane-2,6-dione (18%) as the main product. The use of micelles appears to be not only a new way to synthesize regioisomeric ferulic acid dehydrodimers but may also help to understand the regiospecificity of dimeric hydroxycinnamate formation in vivo.     

Digestion and absorption of ferulic acid sugar esters in rat gastrointestinal tract

We estimated the absorption site and absorptivity of ferulic acid (FA) and its sugar esters, namely 5-O-feruloyl-l-arabinofuranose (FAA) and feruloyl-arabinoxylan (FAXn), in rats on the basis of their recovery in intestinal content and feces by comparing the values with those of a nonabsorbable marker, poly R-478. The results indicated that free FA was absorbed almost completely before reaching cecum. About 40% of dietary FAA was absorbed in rat foregut and 57% disappeared in the cecum. In contrast, about 67% of the FA moiety in FAXn was released and then disappeared predominantly in the hindgut. These results suggested that the existing form of FA in diets affects its absorptivity, its absorption site, and its ensuing fate in the gastrointestinal tract. Those ingested FAs esterified with saccharides; especially, polysaccharides have to transit the hindgut where FA might be released and then absorbed and/or degraded by microflora in lumen. Such microbial degradation may be an important factor affecting the bioavailability of dietary FA.     

A spectrophotometric study of the copigmentation of malvin with caffeic and ferulic acids

The process of copigmentation of the anthocyanin molecule malvidin 3, 5-diglucoside with two organic monocarboxylic phenolic acids, caffeic and ferulic acids, was studied via their absorption electronic spectra. The dependence of the copigmentation process on the pH of the medium, molecular concentration, and temperature was established. The process of copigmentation was observed at two pH values: 2.50 and 3.65. The stoichiometric ratio was 1:1 at both pH values. The copigmentation was characterized by approximately equal values of the equilibrium constant, K, within each of the pH values. The temperature was found to be a significant factor that determines the thermodynamic conditions of the copigmentation process, because the process is spontaneous (DeltaG degrees < 0), and results in entropy loss (DeltaS degrees < 0) at both pH values.     

Peroxidase-mediated cross-linking of a tyrosine-containing peptide with ferulic acid

The tyrosine-containing peptide Gly-Tyr-Gly (GYG) was oxidatively cross-linked by horseradish peroxidase in the presence of hydrogen peroxide. As products, covalently coupled di- to pentamers of the peptide were identified by LC-MS. Oxidative cross-linking of ferulic acid with horseradish peroxidase and hydrogen peroxide resulted in the formation of dehydrodimers. Kinetic studies of conversion rates of either the peptide or ferulic acid revealed conditions that allow formation of heteroadducts of GYG and ferulic acid. To a GYG-containing incubation mixture was added ferulic acid in small aliquots, therewith keeping the molar ratio of the substrates favorable for hetero-cross-linking. This resulted in a predominant product consisting of two ferulic acid molecules dehydrogenatively linked to a single peptide and, furthermore, two ferulic acids linked to peptide oligomers, ranging from dimers to pentamers. Also, mono- and dimers of the peptide were linked to one molecule of ferulic acid. A mechanism explaining the formation of all these products is proposed.     

Analysis of phenolic acids in barley by high-performance liquid chromatography

Phenolic acids from 30 barley varieties (combination of hulled/hulless/two-row/six-row/regular/waxy) were investigated by HPLC following four different sample treatments: (a) simple hot water extraction, (b) extraction after acid hydrolysis, (c) acid plus alpha-amylase hydrolysis, and (d) acid plus alpha-amylase plus cellulase hydrolysis treatments. The benzoic acid (p-hydroxybenzoic, vanillic, and protocatechuic acids) and cinnamic acid derivatives (coumaric, caffeic, ferulic, and chlorogenic acids) were identified, and some of the phenolic acids were quantified after each above-mentioned treatment. The data indicated that a combination of sequential acid, alpha-amylase, and cellulase hydrolysis treatments might be applicable for release of more phenolic acids from barley.     

Hydroxycinnamic acid amide derivatives, phenolic compounds and antioxidant activities of extracts of pollen samples from Southeast Brazil

Seven bee pollen samples (C1-C7) with different palynological sources were harvested from Pindamonhangaba municipality (Southeast Brazil). Methanol extracts of untreated samples (control), samples frozen at -18 °C and samples frozen and then dried were analyzed by HPLC/PAD/ESI/MS/MS. Flavonoid diglycosides of quercetin, kaempferol, isorhamnetin and patuletin were detected, together with hydroxycinnamic acid amide derivatives, such as N',N',N'-tris-p-feruloylspermidine and N',N',N'-tris-p-coumaroylspermidine. Distinct phenolic profiles characterized the analyzed samples, but no differences were noted as resulting from different treatments. Total phenolic contents determined with the Folin-Ciocalteau reagent ranged from 1.7 to 2.2%. Antioxidant activities above 75%, based on the DPPH method, were observed for all extracts, not correlated with total phenolic contents. Among samples from the same origin, those frozen were more active than samples untreated (control), and the samples frozen and then dried were the most active.     

Screening spring wheat for midge resistance in relation to ferulic acid content

The concentration of ferulic acid (FA), the major phenolic acid in the wheat kernel, was found to differ significantly in the mature grain of six wheat cultivars known to have a range of tolerance to the orange wheat blossom midge (Sitodiplosis mosellana). Differences in FA content were correlated with floret infestation level of the cultivars. The wheat cultivars ranked similarly in FA content at the four locations where they were tested, despite a significant effect of environment. Ferulic acid was synthesized mainly during the early stages of grain filling but at different rates among cultivars. Ferulic acid was concentrated primarily in the shorts and bran fractions in an insoluble-bound form. A high correlation was obtained between FA contents as determined by GLC, fluorometry, UV, and colorimetry. The colorimetric procedure was modified as a qualitative, simple, and rapid test for identifying midge-resistant wheat and evaluated in several field trials. The method should provide a rapid tool in the preliminary screening of experimental lines in the development of midge-resistant wheat cultivars.     

Flavanol and bound phenolic acid contents in different barley varieties

Different barley varieties, consisting of hulled and hull-less types, of normal, waxy, and high amylose starch, as well as two-rowed and six-rowed types, were analyzed for their main proanthocyanidins and bound phenolic acids. Variations in proanthocyanidin and phenolic acid contents were studied in different barley types as well as inter-relationships between the phytochemicals and polysaccharides. The main flavanols found in the analyzed barley varieties were two dimeric as well as four trimeric forms in addition to catechin. The total amount of flavanols ranged from 325 to 527 microg/g of fresh weight of barley flour. No evident associations were found between variations in proanthocyanidin levels and different barley types. The total amount of phenolic acids ranged from 604 to 1346 microg/g of fresh weight of barley flour, with ferulic acid as the dominating acid. The amount of phenolic acids varied according to occurrence or lack of hull, with significantly higher levels in the hulled varieties.