Determination of total antioxidant capacity by oxygen radical absorbance capacity (ORAC) using fluorescein as the fluorescence probe: First Action 2012.23.

An improved method for the measurement of oxygen radical absorbance capacity (ORAC) was developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H], 9'[9H]-xanthen]-3-one) as a new fluorescence probe (ORAC(FL)). Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water-solubility enhancer for lipophilic antioxidants. 7% RMCD (w/v) in 50% acetone-H2O mixture sufficiently solubilized vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). Results indicated that fluorescein shows excellent photostability under the plate reader conditions. This ORAC(FL) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrated that the ORACFL assay is reliable and robust. The mean of intraday and interday CVs were <15%; for hydrophilic ORAC, LOD and LOQ are 5 and 6.25 microM, respectively; for lipophilic ORAC, LOD and LOQ are 6.25 and 12.5 microM, respectively. It is concluded that unlike other popular methods, the ORAC(FL) assay provides a direct measure of total antioxidant capacity against the peroxyl radicals.

Determination of total procyanidins in selected chocolate and confectionery products using DMAC.

A simple, specific, high-throughput colorimetric method based on the reaction of 4-dimethylaminocinnamaldehyde (DMAC) with flavan-3-ols was developed to determine total procyanidins in selected cacao-based products. Extracts of defatted samples were dispensed into a 96-well plate and reacted with DMAC. The absorbance of the reaction products was measured at 640 nm and compared to commercially available procyanidin B2 as a standard. The use of the 96-well plates and a plate reader dramatically improved sample throughput. A standard protocol was established and used for further studies. The calibration was found to be linear from 1-100 ppm. The DMAC reagent reacted relatively specifically to (-)-epicatechin, (+)-catechin, epigallocatechin, gallocatechin, the gallates of catechin, epicatechin, gallocatechin, and epigallocatechin, oligomeric procyanidins of cocoa up to n=4, and A-type procyanidins. Little or no reaction occurred with cyanidins and representative compounds of phenolic acids, flavones, flavanones, flavonols, anthocyanidins, isoflavones, and stilbenes. Sample precision studies were carried out on 10 different test materials over several weeks, and yielded RSD values of 4.0 to 9.5%. The method was ring-tested in three laboratories using blinded test materials including cocoa beans, cocoa powder, chocolate liquor, dark chocolate, and milk chocolate. There was excellent agreement of the results between laboratories.

In vitro assessment of oat ß-glucans nutritional properties: An inter-laboratory methodology evaluation.

The purpose of this inter-laboratory study was to test the repeatability and reproducibility of an in vitro method aimed at analyzing the physicochemical properties under physiological conditions of ß-glucans from foods. After evaluating ß-glucans molar mass and quantification methods using five ß-glucan controls, four laboratories ran six oat-based products through in vitro digestion, measured ß-glucans solubility and viscosity and molar mass of solubilized ß-glucans. The determination of the molar mass of ß-glucan controls, their viscosity in solution and ß-glucans content in food samples exhibited relative standard reproducibility of 20.9-40.9%, 10.2-40.9% and 2.3-14.8%, respectively. After in vitro digestion, relative standard reproducibility ranged 12.1-60.0%, 12.2-64.3% and 9.7-36.3% for molar mass of extracted ß-glucans, their viscosity and their solubility, respectively. Although the characterization methods were satisfactory within the limits of current technology, the in vitro extraction contributed significantly to the uncertainty of final characterization.

Rapid quantitation of avenanthramides in oat-containing products by high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-TQMS).

Avenanthramides (AVNs) are a family of nitrogen-containing phenolic compounds produced in oat; AVN 2c, 2p, and 2f are the three major members. An LC-MS/MS method was developed, with the limit of detection (LOD) and the limit of quantitation (LOQ) being, respectively, 0.29ng/mL and 1.96ng/mL for AVN 2c, 0.24ng/mL and 0.60ng/mL for AVE 2p, and 0.42ng/mL and 2.2ng/mL for AVN 2f. The method was validated in oat-containing hot cereal and snack bar samples. The recovery of AVN 2c, 2p, and 2f from these two oat products was 95-113%, and the relative standard deviations ranged from 5% to 9%. This method was used to evaluate oat products and raw oat samples. The effects of location and variety on AVN composition were investigated. The method presented here provides a novel and rapid tool to quantitate the abundance of AVN 2c, 2p, and 2f in oat-containing products.

Procyanidin and catechin contents and antioxidant capacity of cocoa and chocolate products.

Cocoa and chocolate products from major brands were analyzed blind for total antioxidant capacity (AOC) (lipophilic and hydrophilic ORAC(FL)), catechins, and procyanidins (monomer through polymers). Accuracy of analyses was ascertained by comparing analyses on a NIST standard reference chocolate with NIST certified values. Procyanidin (PC) content was related to the nonfat cocoa solid (NFCS) content. The natural cocoa powders (average 87% of NFCS) contained the highest levels of AOC (826 +/- 103 micromol of TE/g) and PCs (40.8 +/- 8.3 mg/g). Alkalized cocoa (Dutched powders, average 80% NFCS) contained lower AOC (402 +/- 6 micromol of TE /g) and PCs (8.9 +/- 2.7 mg/g). Unsweetened chocolates or chocolate liquor (50% NFCS) contained 496 +/- 40 micromol of TE /g of AOC and 22.3 +/- 2.9 mg/g of PCs. Milk chocolates, which contain the least amount of NFCS (7.1%), had the lowest concentrations of AOC (80 +/- 10 micromol of TE /g) and PCs (2.7 +/- 0.5 mg/g). One serving of cocoa (5 g) or chocolate (15 or 40 g, depending upon the type of chocolate) provides 2000-9100 micromol of TE of AOC and 45-517 mg of PCs, amounts that exceed the amount in a serving of the majority of foods consumed in America. The monomers through trimers, which are thought to be directly bioavailable, contributed 30% of the total PCs in chocolates. Hydrophilic antioxidant capacity contributed >90% of AOC in all products. The correlation coefficient between AOC and PCs in chocolates was 0.92, suggesting that PCs are the dominant antioxidants in cocoa and chocolates. These results indicate that NFCS is correlated with AOC and PC in cocoa and chocolate products. Alkalizing dramatically decreased both the procyanidin content and antioxidant capacity, although not to the same extent.

Phytochemical and nutrient composition of the freeze-dried amazonian palm berry, Euterpe oleraceae mart. (acai).

Euterpe oleraceae is a large palm tree indigenous to the Amazon River and its tributaries and estuaries in South America. Its fruit, known as acai, is of great economic value to native people. In this study, a standardized freeze-dried acai fruit pulp/skin powder was used for all analyses and tests. Among many findings, anthocyanins (ACNs), proanthocyanidins (PACs), and other flavonoids were found to be the major phytochemicals. Two ACNs, cyandin 3-glucoside and cyanidin 3-rutinoside were found to be predominant ACNs; three others were also found as minor ACNs. The total content of ACNs was measured as 3.1919 mg/g dry weight (DW). Polymers were found to be the major PACs. The concentration of total PACs was calculated as 12.89 mg/g DW. Other flavonoids, namely, homoorientin, orientin, isovitexin, scoparin, and taxifolin deoxyhexose, along with several unknown flavonoids, were also detected. Resveratrol was found but at a very low concentration. In addition, components including fatty acids, amino acids, sterols, minerals, and other nutrients were analyzed and quantified. Total polyunsaturated fatty acid, total monounsaturated fatty acid, and total saturated fatty acids contributed to 11.1%, 60.2%, and 28.7% of total fatty acid. Oleic acid (53.9%) and palmitic acid (26.7%) were found to be the two dominant fatty acids. Nineteen amino acids were found; the total amino acid content was determined to be 7.59% of total weight. The total sterols accounted for 0.048% by weight of powder. The three sterols B-sitosterol, campesterol, and sigmasterol were identified. A complete nutrient analysis is also presented. Microbiological analysis was also performed.

High-throughput quantitation of peroxyl radical scavenging capacity in bulk oils.

Autoxidation of methyl linoleate (8:2 mixture with decane, 37 degrees C) was induced by 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN, 17.7 mM) and the kinetics of oxygen consumption monitored using a 96-well microplate coated with an oxygen-sensitive fluorescence probe, a ruthenium dye, embedded in a silicone matrix at the bottom of the microplate. The probe does not participate in the reaction; instead, its fluorescence intensity is inversely proportional to the solution oxygen concentration as it changes during oxidation. In the absence of antioxidants, the oxidation rate has a linear relationship with the square root of the initiator concentrations. This is in agreement with theoretical autoxidation kinetics equations. In the presence of tocopherol-type antioxidants, a sharp lag phase appears. The quantitation of the antioxidant capacity is achieved using the area under the curve (AUC) approach. The assay has a 2 h running time, a linearity range from 1.56 to 18.7 microM (Trolox), and a limit of quantitation at 2.7 microM Trolox equivalency. The peroxyl radical scavenging capacities of several cold-pressed and organically grown plant seed oils were quantified along with the tocopherol concentrations of the oils. Tocopherols contribute only a fraction of the peroxyl radical scavenging capacity of the oils, and there is poor correlation between total tocopherol concentrations and radical scavenging capacity, suggesting that the antioxidant capacity of oils is due not only to tocopherols but also to other lipid-soluble antioxidants.

Antioxidant capacity and other bioactivities of the freeze-dried Amazonian palm berry, Euterpe oleraceae mart. (acai).

The fruit of Euterpe oleraceae, commonly known as acai, has been demonstrated to exhibit significantly high antioxidant capacity in vitro, especially for superoxide and peroxyl scavenging, and, therefore, may have possible health benefits. In this study, the antioxidant capacities of freeze-dried acai fruit pulp/skin powder (OptiAcai) were evaluated by different assays with various free radical sources. It was found to have exceptional activity against superoxide in the superoxide scavenging (SOD) assay, the highest of any food reported to date against the peroxyl radical as measured by the oxygen radical absorbance capacity assay with fluorescein as the fluorescent probe (ORACFL), and mild activity against both the peroxynitrite and hydroxyl radical by the peroxynitrite averting capacity (NORAC) and hydroxyl radical averting capacity (HORAC) assays, respectively. The SOD of acai was 1614 units/g, an extremely high scavenging capacity for O2*-, by far the highest of any fruit or vegetable tested to date. Total phenolics were also tested as comparison. In the total antioxidant (TAO) assay, antioxidants in acai were differentiated into "slow-acting" and "fast-acting" components. An assay measuring inhibition of reactive oxygen species (ROS) formation in freshly purified human neutrophils showed that antioxidants in acai are able to enter human cells in a fully functional form and to perform an oxygen quenching function at very low doses. Furthermore, other bioactivities related to anti-inflammation and immune functions were also investigated. Acai was found to be a potential cyclooxygenase (COX)-1 and COX-2 inhibitor. It also showed a weak effect on lipopolysaccharide (LPS)-induced nitric oxide but no effect on either lymphocyte proliferation and phagocytic capacity.

Antioxidant activity and polyphenol and procyanidin contents of selected commercially available cocoa-containing and chocolate products in the United States.

In the United States, commercially available foods, including cocoa and chocolate, are being marketed with statements referring to the level of antioxidant activity and polyphenols. For cocoa-containing foods, there has been no comprehensive survey of the content of these and other chemistries. A survey of cocoa and chocolate-containing products marketed in the United States was conducted to determine antioxidant activity and polyphenol and procyanidin contents. Commercially available samples consisted of the top market share products in each of the following six categories: natural cocoa, unsweetened baking chocolate, dark chocolate, semisweet baking chips, milk chocolate, and chocolate syrup. Composite samples were characterized using four different methods: oxygen radical absorbance capacity (ORAC), vitamin C equivalence antioxidant capacity (VCEAC), total polyphenols, and procyanidins. All composite lots were further characterized for percent nonfat cocoa solids (NFCS) and percent fat. Natural cocoas had the highest levels of antioxidant activities, total polyphenols, and procyanidins followed by baking chocolates, dark chocolates and baking chips, and finally milk chocolate and syrups. The results showed a strong linear correlation between NFCS and ORAC (R (2) = 0.9849), total polyphenols (R (2) = 0.9793), and procyanidins (R (2) = 0.946), respectively. On the basis of principal component analysis, 81.4% of the sample set was associated with NFCS, antioxidant activity, total polyphenols, and procyanidins. The results indicated that, regardless of the product category, NFCS were the primary factor contributing to the level of cocoa antioxidants in the products tested. Results further suggested that differences in cocoa bean blends and processing, with the possible exception of Dutching, are minor factors in determining the level of antioxidants in commercially available cocoa-containing products in the United States.

When east meets west: the relationship between yin-yang and antioxidation-oxidation.

Ancient traditional Chinese medicine (TCM) has effectively relied on the theory of yin-yang balance in diagnoses and treatments of diseases and disorders for more than 2000 years. However, in eastern society, yin-yang is regarded as an incomprehensible ideology without definite physical meaning. Consequently, the yin-yang balance in medicine has not been studied by modern scientific means. In the western world, yin-yang balance is often misunderstood as a religious belief or a principle of lifestyle. Herein, we attempted to define the physical meaning of yin-yang in TCM by correlating it with biochemical processes. We propose that yin-yang balance is antioxidation-oxidation balance with yin representing antioxidation and yang as oxidation. Our proposal is partially supported by the fact that the yin-tonic traditional Chinese herbs have, on average, about six times more antioxidant activity and polyphenolic contents than the yang-tonic herbs. Our hypothesis opens an avenue to systematically study the yin-yang balance and its health implications with the use of modern biochemical tools.

The chemistry behind antioxidant capacity assays.

This review summarizes the multifaceted aspects of antioxidants and the basic kinetic models of inhibited autoxidation and analyzes the chemical principles of antioxidant capacity assays. Depending upon the reactions involved, these assays can roughly be classified into two types: assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). The majority of HAT-based assays apply a competitive reaction scheme, in which antioxidant and substrate compete for thermally generated peroxyl radicals through the decomposition of azo compounds. These assays include inhibition of induced low-density lipoprotein autoxidation, oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change is correlated with the sample's antioxidant concentrations. ET-based assays include the total phenols assay by Folin-Ciocalteu reagent (FCR), Trolox equivalence antioxidant capacity (TEAC), ferric ion reducing antioxidant power (FRAP), "total antioxidant potential" assay using a Cu(II) complex as an oxidant, and DPPH. In addition, other assays intended to measure a sample's scavenging capacity of biologically relevant oxidants such as singlet oxygen, superoxide anion, peroxynitrite, and hydroxyl radical are also summarized. On the basis of this analysis, it is suggested that the total phenols assay by FCR be used to quantify an antioxidant's reducing capacity and the ORAC assay to quantify peroxyl radical scavenging capacity. To comprehensively study different aspects of antioxidants, validated and specific assays are needed in addition to these two commonly accepted assays.

Functional beverage of Garcinia mangostana (mangosteen) enhances plasma antioxidant capacity in healthy adults.

This study was to investigate the absorption and antioxidant effect of a mangosteen-based functional beverage in humans. The beverage contained mangosteen, aloe vera, green tea, and multivitamins. A randomized, double-blind, placebo-controlled clinical trial was conducted with generally healthy male and female subjects between 18 and 60 years of age. Ten men and 10 women participated in this study. Participants were randomly divided into two groups, treatment and placebo group. Participants received either a daily single dose (245 mL) of the beverage or a placebo. Blood samples were collected from each participant at time points 0, 1, 2, 4, and 6 h. The plasma samples were analyzed by LC/MS for α-mangostin and vitamins B2 and B5. Results indicated that the three analytes were bioavailable, with observed C max at around 1 h. The antioxidant capacity measured with the oxygen radical absorbance capacity (ORAC) assay was increased with a maximum effect of 60% after 1 h, and the elevated antioxidant level lasted at least 6 h. This study demonstrated the bioavailability of α-mangostin and B vitamins from a xanthone-rich beverage and the mechanisms of the increase in plasma antioxidant may be direct effects from antioxidants, enhancement of endogenous antioxidant activity through activation of Nrf2 pathway, and synergism of the antioxidants.

Daily consumption of a mangosteen-based drink improves in vivo antioxidant and anti-inflammatory biomarkers in healthy adults: a randomized, double-blind, placebo-controlled clinical trial.

Mangosteen (Garcinia mangostana) is a tropical fruit cultivated mainly in Southeast Asia. Recent studies have shown mangosteen has many health benefits. In this study, we aimed to determine the effects of a mangosteen-based beverage on antioxidant and anti-inflammatory and immunity biomarkers in plasma of healthy adults. A randomized, double-blind, placebo-controlled clinical trial was conducted using 60 participants, 30 men, and 30 women, ages 18-60. Participants were randomly divided into two groups, placebo and mangosteen groups, with the same number of male and female participants in each group. The trial duration was 30 days. ORAC as an antioxidant biomarker was measured in both groups. It was found that after the 30-day trial, the group given the mangosteen-based drink formula showed 15% more antioxidant capacity in the bloodstream than did the placebo group. As for the inflammatory biomarkers, in the mangosteen group, between the preintervention and postintervention, the C-reactive protein level significantly decreased by 46%, while no significant decreases for the same biomarker was observed in the placebo group. Immunity biomarkers IgA, IgG, IgM, C3 and C4 were not affected in either group. In addition, the effects on hepatic function (Aspartate Aminotransferase and Alanine Aminotransferase) and kidney function (creatinine) were investigated. Our results indicated that after the 30-day consumption of the beverage, there were no side effects on human hepatic and kidney functions. The outcome of this study showed that the mangosteen-based formula significantly increases antioxidant capacity and possesses anti-inflammatory benefits with no side effects on immune, hepatic, and renal functions for long-term consumption.

Antioxidant activity and phenolic content of Oregon caneberries.

Five types of caneberries [evergreen blackberries (Rubus laciniatus), marionberries (Rubus ursinus), boysenberries (Rubus ursinus x idaeus), red raspberries (Rubus idaeus), and black raspberries (Rubus occidentalis)] were analyzed for antioxidant activity by measuring their oxygen radical absorbance capacity (ORAC). In addition, the berries were analyzed for total phenolics, anthocyanins, procyanidins, and ellagic acid content. All of the berries had high ORAC activity ranging from 24 to 77.2 micromol of Trolox equiv/g of fresh berries. Anthocyanin content ranged from 0.65 to 5.89 mg/g, and phenolics ranged from 4.95 to 9.8 mg/g. Black raspberries had the highest ORAC and anthocyanin and phenolic contents. Only red raspberries had detectable amounts of procyanidin oligomers (monomer, dimers, and trimers). All berries had high levels of ellagic acid (47-90 mg/g), but boysenberries had the highest level prior to hydrolysis. The results from this study indicate that these caneberries were high in antioxidant activity and were rich sources of anthocyanins and phenolics.

Anthocyanin and proanthocyanidin content in selected white and red wines. Oxygen radical absorbance capacity comparison with nontraditional wines obtained from highbush blueberry.

Antioxidant capacity, as measured by oxygen radical absorbance capacity (ORAC(PE)), total phenolic, total and individual anthocyanins, and proanthocyanidin fraction contents were evaluated in red and white wines from grapes. A comparison in terms of antioxidant capacity is made with nontraditional wines made from highbush blueberry. Blueberries are among fruits that are best recognized for their potential health benefits. In red wines, total oligomeric proanthocyanidin content, including catechins, was substantially higher (177.18 +/- 96.06 mg/L) than that in white wines (8.75 +/- 4.53 mg/L). A relative high correlation in red wines was found between ORAC(PE) values and malvidin compounds (r = 0.75, P < 0.10), and proanthocyanidins (r = 0.87, P < 0.05). In white wines, a significant correlation was found between the trimeric proanthocyanidin fraction and peroxyl radical scavenging values (r = 0.86, P < 0.10). A moderate drink (1 drink per day, about 140 mL) of red wine, or white wine, or wine made from highbush blueberry corresponds to an intake of 2.04 +/- 0.81 mmol of TE, 0.47 +/- 0.15 mmol of TE, and 2.42 +/- 0.88 mmol of TE of ORAC(PE)/day, respectively.

Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe.

A novel fluorometric method has been developed to evaluate hydroxyl radical prevention capacity using fluorescein (FL) as the probe. The hydroxyl radical is generated by a Co(II)-mediated Fenton-like reaction, and the hydroxyl radical formation under the experimental condition is indirectly confirmed by the hydroxylation of p-hydroxybenzoic acid. The fluorescence decay curve of FL is monitored in the absence or presence of antioxidant, the area under the fluorescence decay curve (AUC) is integrated, and the net AUC, which is an index of the hydroxyl radical prevention capacity, is calculated by subtracting the AUC of the blank from that of the antioxidant. Gallic acid is chosen as a reference standard, and the activity of sample is expressed as gallic acid equivalents. The method is rigorously validated through linearity, precision, accuracy, and ruggedness. A wide range of phenolic antioxidants is analyzed, and the hydroxyl radical prevention capacity is mainly due to the metal-chelating capability of the compounds.

Analysis of antioxidant activities of common vegetables employing oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays: a comparative study.

A total of 927 freeze-dried vegetable samples, including 111 white cabbages, 59 carrots, 51 snap beans, 57 cauliflower, 33 white onions, 48 purple onions, 130 broccoli, 169 tomatoes, 25 beets, 88 peas, 88 spinach, 18 red peppers, and 50 green peppers, were analyzed using the oxygen radical absorption capacity (ORAC) and ferric reducing antioxidant capacity (FRAP) methods. The data show that the ORAC and FRAP values of vegetable are not only dependent on species, but also highly dependent on geographical origin and harvest time. The two antioxidant assay methods, ORAC and FRAP, also give different antioxidant activity trends. The discrepancy is extensively discussed based on the chemistry principles upon which these methods are built, and it is concluded that the ORAC method is chemically more relevant to chain-breaking antioxidants activity, while the FRAP has some drawbacks such as interference, reaction kinetics, and quantitation methods. On the basis of the ORAC results, green pepper, spinach, purple onion, broccoli, beet, and cauliflower are the leading sources of antioxidant activities against the peroxyl radicals.

High-throughput assay of oxygen radical absorbance capacity (ORAC) using a multichannel liquid handling system coupled with a microplate fluorescence reader in 96-well format.

The oxygen radical absorbance capacity (ORAC) assay has been widely accepted as a standard tool to measure the antioxidant activity in the nutraceutical, pharmaceutical, and food industries. However, the ORAC assay has been criticized for a lack of accessibility due to the unavailability of the COBAS FARA II analyzer, an instrument discontinued by the manufacturer. In addition, the manual sample preparation is time-consuming and labor-intensive. The objective of this study was to develop a high-throughput instrument platform that can fully automate the ORAC assay procedure. The new instrument platform consists of a robotic eight-channel liquid handling system and a microplate fluorescence reader. By using the high-throughput platform, the efficiency of the assay is improved with at least a 10-fold increase in sample throughput over the current procedure. The mean of intra- and interday CVs was

Development and validation of oxygen radical absorbance capacity assay for lipophilic antioxidants using randomly methylated beta-cyclodextrin as the solubility enhancer.

We recently reported the improved oxygen radical absorbance capacity (ORAC) assay using fluorescein (FL) as the fluorescent probe. The current ORAC(FL) assay is limited in hydrophilic antioxidant due to the aqueous environment of the assay. Lipophilic antioxidants mainly include the vitamin E family and carotenoids, which play a critical role in biological defense systems. In this paper, we expanded the current ORAC(FL) assay to lipophilic antioxidants. Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water solubility enhancer for lipophilic antioxidants. Seven percent RMCD (w/v) in a 50% acetone-H(2)O mixture was found to sufficiently solubilize vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). This newly developed ORAC assay (abbbreviated ORAC(FL-LIPO)) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrate that the ORAC(FL-LIPO) assay is reliable and robust. For the first time, by using 6-hydroxy-2,5,7,8-tetramethyl-2-carboxylic acid as a standard (1.0), the ORAC values of alpha-tocopherol, (+)-gamma-tocopherol, (+)-delta-tocopherol, alpha-tocopherol acetate, tocotrienols, 2,6-di-tert-butyl-4-methylphenol, and gamma-oryzanol were determined to be 0.5 +/- 0.02, 0.74 +/- 0.03, 1.36 +/- 0.14, 0.00, 0.91 +/- 0.04, 0.16 +/- 0.01, and 3.00 +/- 0.26, respectively. The structural information of oxidized alpha-tocopherol obtained by liquid chromatography/mass spectrometry reveals that the mechanism for the reaction between the vitamin E and the peroxyl radical follows the hydrogen atom transfer mechanism, which is in agreement with the notion that vitamin E is the chain-breaking antioxidant.

Chemical composition of caneberry (Rubus spp.) seeds and oils and their antioxidant potential.

Caneberries (Rubus spp. L.) are grown primarily throughout the Pacific Northwestern United States and Canada. Processing of caneberry fruit typically removes the seed, and the development of a value-added use of seeds could expand the market for caneberries and the profit margins for growers. An initial step toward the use of the seeds is a characterization of seed and oil. Our investigation has described compositional characteristics for seeds of five commonly grown caneberry species: red raspberry, black raspberry, boysenberry, Marion blackberry, and evergreen blackberry. Seeds from all five species had 6-7% protein and 11-18% oil. The oils contained 53-63% linoleic acid, 15-31% linolenic acid, and 3-8% saturated fatty acids. The two smaller seeded raspberry species had higher percentages of oil, the lowest amounts of saturated fatty acid, and the highest amounts of linolenic acid. Antioxidant capacities were detected both for whole seeds and for cold-pressed oils but did not correlate to total phenolics or tocopherols. Ellagitannins and free ellagic acid were the main phenolics detected in all five caneberry species and were approximately 3-fold more abundant in the blackberries and the boysenberry than in the raspberries.