Honey with high levels of antioxidants can provide protection to healthy human subjects

Free radicals and reactive oxygen species (ROS) have been implicated in contributing to the processes of aging and disease. Humans protect themselves from these damaging compounds, in part, by absorbing antioxidants from high-antioxidant foods. This report describes the effects of consuming 1.5 g/kg body weight of corn syrup or buckwheat honey on the antioxidant and reducing capacities of plasma in healthy human adults. The corn syrup treatment contained 0.21 +/- 0.06 mg of phenolic antioxidants per gram, and the two buckwheat honey treatments contained 0.79 +/- 0.02 and 1.71 +/- 0.21 mg of phenolic antioxidants per gram. Following consumption of the two honey treatments, plasma total-phenolic content increased (P < 0.05) as did plasma antioxidant and reducing capacities (P < 0.05). These data support the concept that phenolic antioxidants from processed honey are bioavailable, and that they increase antioxidant activity of plasma. It can be speculated that these compounds may augment defenses against oxidative stress and that they might be able to protect humans from oxidative stress. Given that the average sweetener intake by humans is estimated to be in excess of 70 kg per year, the substitution of honey in some foods for traditional sweeteners could result in an enhanced antioxidant defense system in healthy adults.     

Effects of structure on radical-scavenging abilities and antioxidative activities of tea polyphenols: NMR analytical approach using 1,1-diphenyl-2-picrylhydrazyl radicals

Tea (Camellia sinensis) leaves contain various antioxidants such as ascorbic acid (1) and polyphenols. This study tries to clarify the molecular mechanisms underlying the antioxidative and radical-scavenging activities of these antioxidants, and the reactivities of each antioxidant have been compared against that of the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH, 2) using nuclear magnetic resonance (NMR) analysis. Catechol (3) and (+)-taxifolin (4) were oxidized to o-quinone by 2. However, ethyl protocatechuate (5) and quercetin (6) were not oxidized to o-quinone, even though they possess a catechol structure. The radical-scavenging ability of o-dihydroxyl phenolic compounds with a conjugated olefinic double bond (e.g., 6) was superior to that of compounds without this bond (e.g., 4), whereas the ability of o-dihydroxyl phenolic compounds possessing a conjugated carbonyl bond (5) was inferior to that of compounds lacking this bond (3). Vicinal trihydroxyl phenolic compounds with a conjugated olefinic double bond [e.g., myricetin (7)] had an inferior scavenging ability as compared with compounds lacking this bond [e.g., pyrogallol (8)], but 7 was a better scavenger than compounds with a conjugated carbonyl double bond [e.g., ethyl gallate (9)]. In addition, vicinal trihydroxyl phenolic compounds (e.g., 9) were superior to o-dihydroxyl phenolic compounds (e.g., 6). Finally, 1 scavenged radicals more quickly than 8.     

Comparison of radical scavenging effect, inhibition of microsomal oxygen free radical generation, and serum lipoprotein oxidation of several natural antioxidants

Typical components of the Mediterranean diet, such as olive oil and red wine, contain high concentrations of complex phenols, which have been suggested to have an important antioxidant role. The aim of the present work was to determine the inhibitory potency of compounds such as oleuropein, hydroxytyrosol, and other structurally related compounds, such as gallic acid, toward reactive oxygen species generation and free radical scavenging ability. The potency of these compounds was also examined with respect to protecting in vitro low-density lipoprotein oxidation. These studies indicate that complex phenols, such as hydroxytyrosol, and gallic acid both inhibit free radical generation and act as free radical scavengers. The use of three different approaches to determine antioxidant potency demonstrates that activity in one test does not necessarily correlate with activity in another. It was also demonstrated that the presence of two phenolic groups is not always associated with antioxidant activity.     

Antioxidant properties of roasted coffee residues

The antioxidant activity of roasted coffee residues was evaluated. Extraction with four solvents (water, methanol, ethanol, and n-hexane) showed that water extracts of roasted coffee residues (WERCR) produced higher yields and gave better protection for lipid peroxidation. WERCR showed a remarkable protective effect on oxidative damage of protein. In addition, WERCR showed scavenging of free radicals as well as the reducing ability and to bind ferrous ions, indicating that WERCR acts as both primary and secondary antioxidants. The HPLC analyses showed that phenolic acids (chlorogenic acid and caffeic acid) and nonphenolic compounds [caffeine, trigonelline, nicotinic acid, and 5-(hydroxymethyl)furfuraldehyde] remained in roasted coffee residues. These compounds showed a protective effect on a liposome model system. The concentrations of flavonoids and polyphenolic compounds in roasted coffee residues were 8,400 and 20,400 ppm, respectively. In addition, the Maillard reaction products (MRPs) remaining in roasted coffee residues were believed to show antioxidant activity. These data indicate that roasted coffee residues have excellent potential for use as a natural antioxidant source because the antioxidant compounds remained in roasted coffee residues.     

Antioxidant activity of malt rootlet extracts

To improve the malt rootlet value, the antioxidant potentialities of this byproduct of the malting industry have been analyzed. Three extracts have been considered from the points of view of dry matter yield, level of antioxidant compounds, and efficiency and cost of the extraction protocols. They respectively contain rootlet oil, free phenolic compounds, and bound phenolic compounds. The rootlet oil contains only a low quantity of tocopherols (respectively, 20.6 and 4.2 microgram of alpha-tocopherol and gamma-tocopherol per gram of dry rootlets), and a weak antioxidant activity, evaluated in a stripped corn oil by following spectrophotometrically the conjugated dienes, has been pointed out. The bound compound extract presents a good antioxidant power mainly due to the presence of trans-ferulic and trans-p-coumaric acids, but the dry matter yield is low (2%). The free compound extract has a good antioxidant power, and the valuable dry matter, mainly composed of proteins (52%), sugars (33%), and reducing compounds (5.5%), has a yield of 12%. The mixing of bound and free compound extracts presents an antagonistic effect on the antioxidant power, but a synergistic effect has been pointed out for the mixing of alpha-tocopherol and free compound extract.     

Autotrophic and heterotrophic mechanisms for ammonia oxidation

Abstract. Autotrophic nitrifiers such as Nitrosomonas use ammonia mono-oxygenase for the initial stage of ammonia oxidation. Nitrification inhibitors have this enzyme as their site of action. Their mechanisms include alternative substrates, suicide substrates and cuprous copper chelators.
In heterotrophs, organic nitrogen is normally in the fully reduced state, but a few cell metabolites contain N-O bonds. The synthesis and breakdown of such compounds provides a mechanism for heterotrophic nitrification. A non-enzymic mechanism for nitrogen-oxidation involves hydroxyl radicals produced by the Fenton reaction. Heterotrophic nitrification is particularly important in woodland soils, where wood-rotting fungi use free radicals to break down lignin. Tests for a radical mechanism are described.     

Ascorbic acid, phenolic acid, flavonoid, and carotenoid profiles of selected extracts from Ribes nigrum

Small fruits such as berries have low energy contents, but high contents of vitamins, micronutrients, and dietary fibers and constitute a good source of natural antioxidant compounds that are important constituents of the human diet. This study identified a large number of compounds in an extract of black currant showing high antioxidant activity and compared their profile in various parts of the plants (leaves, buds, and berries). If it was known that berries contained very high levels of natural phenolic compounds, this study showed that leaves and buds could also be considered good sources of natural antioxidants. Indeed, they contained high amounts of phenolic acids, flavonoids, and carotenoids. An acetone mixture can extract several classes of phenolic compounds with a good yield of flavonols, flavan-3-ols, and anthocyanins.     

Antioxidant effect of ferulic acid in isolated membranes and intact cells: synergistic interactions with alpha-tocopherol, beta-carotene, and ascorbic acid

Although an antioxidant mechanism has been involved in the beneficial effects of ferulic acid in human diseases, there are few reports on the antioxidant properties of this compound in isolated membranes and intact cells. Here, we evaluated the ability of ferulic acid in inhibiting lipid peroxidation in rat liver microsomal membranes and reactive oxygen species production in NIH-3T3 fibroblasts, induced by both tert-BOOH and AAPH. We also compared its antioxidant efficiency with that of other antioxidants, such as alpha-tocopherol, beta-carotene, and ascorbic acid, added alone or in combination. Ferulic acid acted as a potent antioxidant in our models, being more effective in protecting from tert-BOOH than from AAPH. Moreover, the compound was the most effective among the antioxidants tested. Synergistic interactions were observed when the compound was used in combination with the other antioxidants, suggesting that they can cooperate in preserving physiological integrity of cells exposed to free radicals.     

Soil organic matter chemistry in allophanic soils: a pyrolysis-GC/MS study of a Costa Rican Andosol catena

Soil organic matter (SOM) in allophanic soils is supposed to accumulate due to protection caused by binding to allophane, aluminium and iron. We investigated a catena of allophanic and non‐allophanic soils in Costa Rica to determine the effect of such binding mechanisms on SOM chemistry. These soils contain no contribution of black carbon. Molecular characterization of litter, extractable and dispersed organic matter was done by Curie‐point pyrolysis‐GC/MS. The molecular chemistry of the organic fractions indicates a strong decomposition of plant‐derived organic matter and a strong contribution of microbial sugars and N‐compounds to SOM. Both the decomposition of plant‐derived SOM – including that of relatively recalcitrant compounds – and the relative contribution of microbial SOM were greater in allophanic samples than in non‐allophanic ones. This suggests that chemical protection does not act on primary OM, although it may influence the accumulation of secondary OM in these soils. The effect of allophane on SOM contents in such perhumid soils is probably through incorporation of decomposition products and microbial SOM in very fine aggregates that – in a perhumid environment – remain saturated with water during much of the year. Greater concentrations of aliphatics are found in allophanic residues, but there is no evidence of any specific mineral‐organic binding. The results do not support the existing theory of chemical protection of plant‐derived components through binding to allophane, iron and aluminium.     

Antioxidant activity and partitioning of phenolic acids in bulk and emulsified methyl linoleate.

To evaluate the effect of colloidal parameters on the activity of natural antioxidants, the effect of selected phenolic acids on both bulk and emulsified methyl linoleate oxidation (in the dark at 40 degrees C) was examined. Oxidation was monitored by determining the formation of hydroperoxides; their isomer distribution and ketodiene (oxodiene) products were monitored by using high-performance liquid chromatography. This study showed the system- and concentration-dependent antioxidant activity of phenolic acids. The scavenging of alpha,alpha-diphenyl-beta-picrazylhydrazyl radicals reflected the antioxidant activity in a bulk oil system but not in an emulsion. Specific interactions of the antioxidant with other compounds, for example, the emulsifier, and intramolecular hydrogen bonds may play an important role in reducing the antioxidant activity. Furthermore, these interactions of antioxidants with emulsifier have a strong influence on the partitioning of antioxidants. Thus, the proportion of the antioxidant solubilized in the lipid phase and particularly in the interface did not necessarily reflect the efficiency of the antioxidant.     

Ability of surface-active antioxidants to inhibit lipid oxidation in oil-in-water emulsion

Lipid oxidation in dispersed lipids is prevalent at the oil-water interface where lipid hydroperoxides are decomposed into free radicals by transition metals. Free radical scavenging antioxidants are believed to be most effective in lipid dispersions when they accumulate at the oil-water interface. The surface activity of antioxidants could be increased by their conjugation to hydrocarbon chains. In this study, p-hydroxyphenylacetic acid (HPA) was conjugated with either a butyl or dodecyl group. The HPA conjugates were more effective at decreasing interfacial tension than unconjugated HPA, indicating that they were able to adsorb at lipid-water interfaces. However, free HPA was a more effective antioxidant than butyl and dodecyl conjugates in Menhaden oil-in-water emulsions as determined by both lipid hydroperoxides and thiobarbituric acid reactive substances. The increased antioxidant activity of free HPA could be due to its more effective free radical scavenging activity and its higher concentration in the lipid phase of oil-in-water emulsions in the presence of surfactant micelles where it can act as a chain-breaking antioxidant.     

Screening and identification of antioxidants in biological samples using high-performance liquid chromatography-mass spectrometry and its application on Salacca edulis Reinw

In this study, a new approach was developed for screening and identifying antioxidants in biological samples. The approach was based on significant decreases of the intensities of ion peaks obtained from high-performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) upon reaction with 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals. HPLC-MS/MS was further applied to elucidate structures of antioxidant peaks characterized in a spiking test. The new approach could also be used to monitor the reactivity of antioxidants in biological sample with free radicals. The approach was successfully applied to the identification of antioxidants in salak (Salacca edulis Reinw), a tropical fruit that is reported to be a very good source of natural antioxidants, but it was still not clear which compounds were responsible for its antioxidant property. The antioxidants in salak were identified to be chlorogenic acid, (-)-epicatechin, and singly linked proanthocyanidins that mainly existed as dimers through hexamers of catechin or epicatechin. In salak, chlorogenic acid was identified to be an antioxidant of the slow reaction type as it reacted with free radicals much more slowly than either (-)-epicatechin or proanthocyanidins. The new approach was proved to be useful for the characterization and identification of antioxidants in biological samples as a mass detector combined with an HPLC separation system not only serves as an ideal tool to monitor free radical active components but also provides their possible chemical structures in a biological sample.     

Wastes generated during the storage of extra virgin olive oil as a natural source of phenolic compounds

Phenolic compounds in extra virgin olive oil (EVOO) have been associated with beneficial effects for health. Indeed, these compounds exert strong antiproliferative effects on many pathological processes, which has stimulated chemical characterization of the large quantities of wastes generated during olive oil production. In this investigation, the potential of byproducts generated during storage of EVOO as a natural source of antioxidant compounds has been evaluated using solid-liquid and liquid-liquid extraction processes followed by rapid resolution liquid chromatography (RRLC) coupled to electrospray time-of-flight and ion trap mass spectrometry (TOF/IT-MS). These wastes contain polyphenols belonging to different classes such as phenolic acids and alcohols, secoiridoids, lignans, and flavones. The relationship between phenolic and derived compounds has been tentatively established on the basis of proposed degradation pathways. Finally, qualitative and quantitative characterizations of solid and aqueous wastes suggest that these byproducts can be considered an important natural source of phenolic compounds, mainly hydroxytyrosol, tyrosol, decarboxymethyl oleuropein aglycone, and luteolin, which, after suitable purification, could be used as food antioxidants or as ingredients in nutraceutical products due to their interesting technological and pharmaceutical properties.     

On-line HPLC-DPPH screening method for evaluation of radical scavenging phenols extracted from apples (Malus domestica L.)

An on-line HPLC-DPPH screening method for phenolic antioxidants in apple methanol/water (80:20, v/v) extracts was applied. The determination of antioxidants was based on a decrease in absorbance at 515 nm after postcolumn reaction of HPLC-separated antioxidants with the 2,2'-diphenyl-1-picrylhydrazyl radicals (DPPH*). Each of the antioxidants separated by the HPLC column was observed as a negative peak corresponding to its antioxidative activity. The on-line method was applied for quantitative analysis of the antioxidants. A linear dependence of negative peak area on concentration of the reference antioxidants was observed. For validation of the on-line method the limit of detection, LOD (microg/mL), and the limit of quantification, LOQ (microg/mL), of the phenolic compounds were determined. Comparison of the UV and DPPH radical quenching chromatograms with authentic compounds identified catechin, chlorogenic acid, caffeic acid, epicatechin, and phloridzin in the apple cultivars (Lobo, Golden Delicious, and Boskoop), and the distribution of total antioxidant activity was calculated.     

How relevant is recalcitrance for the stabilization of organic matter in soils?

Traditionally, the selective preservation of certain recalcitrant organic compounds and the formation of recalcitrant humic substances have been regarded as an important mechanism for soil organic matter (SOM) stabilization. Based on a critical overview of available methods and on results from a cooperative research program, this paper evaluates how relevant recalcitrance is for the long‐term stabilization of SOM or its fractions. Methodologically, recalcitrance is difficult to assess, since the persistence of certain SOM fractions or specific compounds may also be caused by other stabilization mechanisms, such as physical protection or chemical interactions with mineral surfaces. If only free particulate SOM obtained from density fractionation is considered, it rarely reaches ages exceeding 50 y. Older light particles have often been identified as charred plant residues or as fossil C. The degradability of the readily bioavailable dissolved or water‐extractable OM fraction is often negatively correlated with its content in aromatic compounds, which therefore has been associated with recalcitrance. But in subsoils, dissolved organic matter aromaticity and biodegradability both are very low, indicating that other factors or compounds limit its degradation. Among the investigated specific compounds, lignin, lipids, and their derivatives have mean turnover times faster or similar as that of bulk SOM. Only a small fraction of the lignin inputs seems to persist in soils and is mainly found in the fine textural size fraction (<20 µm), indicating physico‐chemical stabilization. Compound‐specific analysis of ¹³C : ¹²C ratios of SOM pyrolysis products in soils with C3‐C4 crop changes revealed no compounds with mean residence times of > 40–50 y, unless fossil C was present in substantial amounts, as at a site exposed to lignite inputs in the past. Here, turnover of pyrolysis products seemed to be much longer, even for those attributed to carbohydrates or proteins. Apparently, fossil C from lignite coal is also utilized by soil organisms, which is further evidenced by low ¹⁴C concentrations in microbial phospholipid fatty acids from this site. Also, black C from charred plant materials was susceptible to microbial degradation in a short‐term (60 d) and a long‐term (2 y) incubation experiment. This degradation was enhanced, when glucose was supplied as an easily available microbial substrate. Similarly, SOM mineralization in many soils generally increased after addition of carbohydrates, amino acids, or simple organic acids, thus indicating that stability may also be caused by substrate limitations. It is concluded that the presented results do not provide much evidence that the selective preservation of recalcitrant primary biogenic compounds is a major SOM‐stabilization mechanism. Old SOM fractions with slow turnover rates were generally only found in association with soil minerals. The only not mineral‐associated SOM components that may be persistent in soils appear to be black and fossil C.     

Novel antioxidant reactions of cinnamates in wine

Plant-derived polyphenolic compounds have received much attention for their ability to sequester high-energy free radicals in a great variety of food-related and biological systems, protecting those systems from oxidative change. The ability of these compounds to scavenge free radicals has always been attributed to their phenolic functionality, from which a hydrogen atom can be easily abstracted. In this study, the cinnamates and the ubiquitous hydroxycinnamates were found to equally suppress the formation of oxidation products in wine exposed to the Fenton reaction (catalytic Fe(II) with hydrogen peroxide). Mechanistic investigations led to the unexpected discovery that the α,β-unsaturated side chain of cinnamic acids could efficiently trap 1-hydroxyethyl radicals, representing a newly discovered mode of antioxidant radical scavenging activity for these broadly occurring compounds in a food system. The proposed pathway is supported by prior fundamental studies with radiolytically generated radicals.     

Total oxidant scavenging capacity of Euterpe oleracea Mart. (açaí) seeds and identification of their polyphenolic compounds

The antioxidant capacity of methanol and ethanol seed extracts from Euterpe oleracea Mart. (a?aí) against the reactive oxygen species (ROS) peroxyl radicals, peroxynitrite, and hydroxyl radicals was studied with the total oxidant scavenging capacity (TOSC) assay in a modified and automated version. Cold methanol digestion was the most efficient extraction method with respect to the antioxidant capacity. The extracts exhibit good antioxidant capacity against peroxyl radicals, similar to the capacity of the pulp. The antioxidant capacity against peroxynitrite and hydroxyl radicals is even higher. The main antioxidants identified by HPLC-MS and HPLC-CEAD are five different procyanidins (di- through pentamers); furthermore, protocatechuic acid and epicatechin were identified as minor compounds. Determination of TOSC values of HPLC seed extract fractions indicates that the procyanidins contribute substantially to the overall antioxidant capacity. In addition, however, other compounds that have not yet been identified are responsible for a large part of the observed antioxidant capacity.     

Effects of solid-state enzymatic treatments on the antioxidant properties of wheat bran

This study evaluated the potential of solid-state enzyme treatments to release insoluble bound antioxidants such as phenolic acids from wheat bran, thereby improving its extractable and potentially bioaccessible antioxidant properties including scavenging capacities against peroxyl (ORAC), ABTS cation, DPPH and hydroxyl radicals, total phenolic contents, and phenolic acid compositions. Investigated enzyme preparations included Viscozyme L, Pectinex 3XL, Ultraflo L, Flavourzyme 500L, Celluclast 1.5L, and porcine liver esterase. Results showed significant dose-dependent increases in extractable antioxidant properties for some enzyme preparations, and Ultraflo L was found to be the most efficient enzyme, able to convert as much as 50% of the insoluble bound ferulic acid in wheat bran to the soluble free form. The effect of moisture content on these solid-state enzyme reactions was also evaluated and found to be dependent on enzyme concentration. These data suggest that solid-state enzyme treatments of wheat bran may be a commercially viable post-harvest procedure for improving the bioaccessibility of wheat antioxidants.     

LDL isolated from plasma-loaded red wine procyanidins resist lipid oxidation and tocopherol depletion

Dietary phenolic compounds may act as antioxidants in vitro, but because of structural modifications during absorption, its role based on concentrations high enough to afford an antioxidant protection needs to be re-evaluated. We have explored the hypothesis that red wine procyanidins interact with low density lipoproteins (LDL) and that, at this location, the phenolic compounds efficiently protect LDL from oxidation and maintain LDL alpha-tocopherol at a high steady state concentration by recycling it back from the alpha-tocopheroxyl radical. To this end, human plasma was supplemented with wine procyanidins and isolated LDL were challenged with a constant flux of peroxyl radicals. As compared with LDL from plasma-free procyanidins, those LDL better resisted lipid oxidation and exhibited longer lag-phases of alpha-tocopherol consumption. The procyanidins, depending on their structure, were able to reduce the UV-induced alpha-tocopherol radical in a micellar system, as evidenced by electron paramagnetic ressonance. Mechanistically, the protection of LDL was interpreted in terms of quenching of peroxyl radicals and the recycling of alpha-tocopherol by the procyanidins bound to the lipoproteins. These results support the notion that, in human plasma, the procyanidins, via binding to LDL, may act as efficient local antioxidants.     

Antioxidant content and free radical scavenging ability of fresh red pummelo [Citrus grandis (L.) Osbeck] juice and freeze-dried products

The antioxidative phytochemicals in various fruits and vegetables are widely recognized for their role in scavenging free radicals, which are involved in the etiology of many chronic diseases. Colored fruits are especially considered a quality trait that correlates with their nutritional values and health benefits. The specific aim of this study was to investigate the antioxidants in the juice and freeze-dried flesh and peel of red pummelo and their ability to scavenge free radicals and compare them with those in white pummelo juice. The total phenolic content of red pummelo juice extracted by methanol (8.3 mg/mL) was found to be significantly higher than that of white pummelo juice (5.6 mg/mL). The carotenoid content of red pummelo juice was also significantly higher than that in white pummelo juice. The contents of vitamin C and delta-tocopherol in red pummelo juice were 472 and 0.35 mug/mL, respectively. The ability of the antioxidants found in red pummelo juice to scavenge radicals were found by methanol extraction to approximate that of BHA and vitamin C with a rapid rate in a kinetic model. The ability of methanol extracts of freeze-dried peel and flesh from red pummelo to scavenge these radicals was 20-40% that of BHA and vitamin C effects. Fresh red pummelo juice is an excellent source of antioxidant compounds and exhibited great efficiency in scavenging different forms of free radicals including DPPH, superoxide anion, and hydrogen peroxide radicals.