Rapid peroxyl radical scavenging capacity (PSC) assay for assessing both hydrophilic and lipophilic antioxidants

This paper reports a simple, rapid, and sensitive assay for assessing peroxyl radical scavenging capacity (PSC) of both hydrophilic and lipophilic antioxidant compounds and food extracts. The assay is based on the degree of inhibition of dichlorofluorescin oxidation by antioxidants that scavenge peroxyl radicals, generated from thermal degradation of 2,2'-azobis(amidinopropane). For hydrophilic antioxidant activity, the dose required to cause a 50% inhibition of the reaction (EC(50)) ranged from 2.41 +/- 0.02 (EGCG) to 21.26 +/- 0.38 microM (ferulic acid). EC(50) values for the hydrophilic antioxidant activity of food extracts ranged from 309.2 +/- 3.63 (apple) to 3345.1 +/- 151.5 micromol of vitamin C equiv/100 g for wheat bran. The EC(50) values for lipophilic antioxidant activity were 1.58 +/- 0.11 (Trolox), 4.35 +/- 0.43 (alpha-tocopherol), 18.94 +/- 0.38 (BHA), and 182.69 +/- 13.7 microM (BHT). Whole grain lipophilic antioxidant activity ranged from 3.49 +/- 0.57 (wheat) to 8.79 +/- 1.98 micromol of alpha-tocopherol equiv/100 g of rice. Hydrophilic antioxidant activity contributed >98% of the total antioxidant activity (hydrophilic plus lipophilic) of whole grains tested. The PSC assay was accurate (86-108% recovery), precise (0.12-11% CV), and reproducible (12% RSD) and produced results comparable to those of similar published assays. The PSC assay can be routinely used to analyze or screen both hydrophilic and lipophilic antioxidants or food extracts and will be a valuable alternative biomarker for future epidemiological studies of chronic diseases.     

Phytochemical profiles and antioxidant activity of wheat varieties

Whole grain consumption has been associated with reduced risk of chronic diseases, such as cardiovascular diseases and cancer. These beneficial effects have been attributed to the unique phytochemicals of grains that complement those found in fruits and vegetables. Wheat is one of the major grains in the human diet; however, little is known about the inherent varietal differences in phytochemical profiles, total phenolic and carotenoid contents, or total antioxidant activities of different wheat varieties, which ultimately influence the associated nutritional and health benefits of wheat and wheat products. The objectives of this study were to determine the phytochemical profiles and total antioxidant activity for 11 diverse wheat varieties and experimental lines. The profiles included free, soluble-conjugated, and insoluble-bound forms of total phenolics, flavonoids, and ferulic acids and carotenoid content including lutein, zeaxanthin, and beta-cryptoxanthin. The results showed that total phenolic content (709.8-860.0 micromol of gallic acid equiv/100 g of wheat), total antioxidant activity (37.6-46.4 micromol of vitamin C/g), and total flavonoid content (105.8-141.8 micromol of catechin equiv/100 g of wheat) did not vary greatly among the 11 wheat lines. However, significant differences in total ferulic acid content (p < 0.05) and carotenoid content (p < 0.05) among the varieties were observed, with carotenoid content exhibiting the greatest range of values. Carotenoid content among the 11 wheat varieties exhibited 5-fold, 3-fold, and 12-fold differences in lutein, zeaxanthin, and beta-cryptoxanthin, respectively. A synthetic wheat experimental line, W7985, gave the lowest carotenoid concentrations of any of the genotypes in this study. Such large genotypic differences in carotenoid content may open up new opportunities for breeding wheat varieties with higher nutritional value.     

Tocopherol and Tocotrienol Content in Commercial Wheat Mill Streams

The content of tocopherols and tocotrienols, collectively known as vitamin E (tocols), was determined in fractions of roller‐milled wheat grains. The results showed that vitamin E components are present in all major flour fractions of wheat, but that the vitamin E content and composition differed significantly between fractions. The total content of vitamin E, calculated as alpha‐tocopherol equivalents, changed from 16.1 mg α‐TE/g in wheat grain to 12.2 mg α‐TE/g in roller‐milled wheat flour. The germ fraction had the highest content of tocopherols, and the content of α‐tocopherol (195.2 μg/g) was 16 times higher (on average) than in any other fraction. The content of tocotrienols was distributed more uniform in the wheat grain with the highest content in the bran fractions, and the content of β‐tocotrienol was higher than the content of α‐tocopherol in all milling fractions except the wheat germ. The content of β‐tocotrienol was 24.1 μg/g in wheat grain, 25.3–31.0 μg/g in the bran fractions, and 14.3–21.9 μg/g in the fractions of endosperm. Overall, germ and fine bran fractions represent good sources of vitamin E and might be used in breadmaking.     

Stability of Vitamin E in Wheat Flour and Whole Wheat Flour During Storage

The stability of vitamin E during 297 days of storage of wheat flour and whole wheat flour ground on a stone mill or a roller mill, respectively, were studied. One day after milling, the total content of vitamin E, expressed in vitamin E equivalents (α‐TE), was 18.7 α‐TE and 10.8 α‐TE for stone‐milled and roller‐milled wheat flour, respectively. The difference in total vitamin E content was primarily due to the absence of the germ and bran fractions in the roller‐milled flour. The total loss of vitamin E during storage was 24% for stone‐milled wheat flour but 50% for roller‐milled wheat flour. These results indicate that vitamin E, which is present in high amounts in wheat germ, functions as an antioxidant in the stone‐milled wheat flour. Hexanal formation showed that lipid oxidation in roller‐milled flour occurred just after milling, whereas the formation of hexanal in the germ fraction displayed a lack period of 22 days, confirming that vitamin E functions as an effective antioxidant in the wheat germ. Results showed no significant difference in total loss of vitamin E for stone‐milled and roller‐milled whole wheat flour. Total loss after 297 days of storage for both milling methods was ≈32%.     

Antioxidant activity of grains

Epidemiological studies have shown that consumption of whole grains and grain-based products is associated with reduced risk of chronic diseases. The health benefits of whole grains are attributed in part to their unique phytochemical composition. However, the phytochemical contents in grains have been commonly underestimated in the literature, because bound phytochemicals were not included. This study was designed to investigate the complete phytochemical profiles in free, soluble conjugated, and insoluble bound forms, as well as their antioxidant activities in uncooked whole grains. Corn had the highest total phenolic content (15.55 +/- 0.60 micromol of gallic acid equiv/g of grain) of the grains tested, followed by wheat (7.99 +/- 0.39 micromol of gallic acid equiv/g of grain), oats (6.53 +/- 0.19 micromol of gallic acid equiv/g of grain), and rice (5.56 +/- 0.17 micromol of gallic acid equiv/g of grain). The major portion of phenolics in grains existed in the bound form (85% in corn, 75% in oats and wheat, and 62% in rice), although free phenolics were frequently reported in the literature. Ferulic acid was the major phenolic compound in grains tested, with free, soluble-conjugated, and bound ferulic acids present in the ratio 0.1:1:100. Corn had the highest total antioxidant activity (181.42 +/- 0.86 micromol of vitamin C equiv/g of grain), followed by wheat (76.70 +/- 1.38 micromol of vitamin C equiv/g of grain), oats (74.67 +/- 1.49 micromol of vitamin C equiv/g of grain), and rice (55.77 +/- 1.62 micromol of vitamin C equiv/g of grain). Bound phytochemicals were the major contributors to the total antioxidant activity: 90% in wheat, 87% in corn, 71% in rice, and 58% in oats. Bound phytochemicals could survive stomach and intestinal digestion to reach the colon. This may partly explain the mechanism of grain consumption in the prevention of colon cancer, other digestive cancers, breast cancer, and prostate cancer, which is supported by epidemiological studies.     

Phytochemical quantification and total antioxidant capacities of emmer (Triticum dicoccon Schrank) and einkorn (Triticum monococcum L.) wheat landraces

In this study, samples of 18 ancient wheat (12 emmer, 6 einkorn) and 2 bread wheat varieties grown in different regions of Turkey were examined for their total phenolics and flavonoids, phenolic acids, lutein, total yellow pigment, and total radical scavenging capacities against ABTS cation. Results showed that health beneficial phytochemicals and total antioxidant capacities were generally significantly different in emmer and einkorn wheat groups. Remarkably higher total antioxidant activity (18.31 +/- 1.31 micromol Trolox equiv/g), total phenolics (6.33 +/- 0.98 micromol gallic acid equiv/g), ferulic acid (662.95 +/- 61.07 microg/g), and flavonoids (1.61 +/- 0.34 micromol catechin equiv/g) content were detected in emmer wheat samples (n = 12), suggesting that they may have high potential for utilization as a novel grain, rich in natural antioxidants. In addition, quite high levels of lutein (7.33 +/- 2.43 microg/g) of einkorn samples (n = 6) hold the potential of developing high-lutein bakery products to considerably raise the dietary intake of carotenoids. These findings for ancient wheat varieties are considered to be very useful in breeding programs for selecting and breeding wheat varieties for higher concentration and better composition of health-beneficial phytochemicals.     

Phenolic acid, tocopherol and carotenoid compositions, and antioxidant functions of hard red winter wheat bran

An electron spin resonance (ESR) spectrometry study was conducted to examine the free radical scavenging properties of bran extracts of Alliance and Wichita wheat using hydroxyl radical (HO*), 2,2-diphenyl-1-picryhydrazyl radical (DPPH*), and superoxide radical anion (O2*-) and their chelating capacities against Cu2+. Also reported is the radical cation 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS*+) scavenging activity, oxygen radical absorbing capacity (ORAC), and chelating property against Fe2+ of the bran extracts measured by the spectrophotometric methods. Significant radical scavenging and chelating capacities were detected in the bran extracts, along with significant levels of phenolic acids, tocopherols, and carotenoids. Ferulic acid, with a concentration range of 130.60-146.38 microg/g, was the predominant phenolic acid in all of the tested bran samples and accounted for approximately 53-67% of total phenolic acids on a weight basis. Total tocopherol concentration ranged from 1.87 to 2.95 micromol/100 g of bran, whereas total carotenoid level was 0.20-0.33 micromol/100 g of bran. In addition, both wheat variety and growing conditions might significantly alter antioxidant properties and concentrations of beneficial components in wheat bran.     

Dry‐Milling and Fractionation of Transgenic Maize Seed Tissues with Green Fluorescent Protein as a Tissue Marker

The efficiency of fractionating cereal grains (e.g., dry corn milling) can be evaluated and monitored by quantifying the proportions of seed tissues in each of the recovered fractions. The quantities of individual tissues are typically estimated using indirect methods such as quantifying fiber or ash to indicate pericarp and tip cap contents, and oil to indicate germ content. More direct and reliable methods are possible with tissue‐specific markers. We used two transgenic maize lines, one containing the fluorescent protein green fluorescent protein (GFP) variant S65T expressed in endosperm, and the other containing GFP expressed in germ to determine the fate of each tissue in the dry‐milling fractionation process. The two lines were dry‐milled to produce three fractions (bran‐, endosperm‐, and germ‐rich fractions) and GFP fluorescence was quantified in each fraction to estimate the tissue composition. Using a simplified laboratory dry‐milling procedure and our GFP‐containing grain, we determined that the endosperm‐rich fraction contained 4% germ tissue, the germ‐rich fraction contained 28% germ, 20% endosperm, and 52% nonendosperm and nonembryo tissues, and the bran‐rich fraction contained 44% endosperm, 13% germ, and 43% nonendosperm and nonembryo tissues. GFP‐containing grain can be used to optimize existing fractionation methods and to develop improved processing strategies.     

Phytochemical profile of main antioxidants in different fractions of purple and blue wheat, and black barley

Two pigmented wheat genotypes (blue and purple) and two black barley genotypes were fractionated in bran and flour fractions, examined, and compared for their free radical scavenging properties against 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (Trolox equivalent antioxidant capacity, TEAC), ferric reducing antioxidant power (FRAP), total phenolic content (TPC), phenolic acid composition, carotenoid composition, and total anthocyanin content. The results showed that fractionation has a significant influence on the antioxidant properties, TPC, anthocyanin and carotenoid contents, and phenolic acid composition. Bran fractions had the greatest antioxidant activities (1.9-2.3 mmol TEAC/100 g) in all four grain genotypes and were 3-5-fold higher than the respective flour fractions (0.4-0.7 mmol TEAC/100 g). Ferulic acid was the predominant phenolic acid in wheat genotypes (bran fractions) while p-coumaric acid was the predominant phenolic acid in the bran fractions of barley genotypes. High-performance liquid chromatography analysis detected the presence of lutein and zeaxanthin in all fractions with different distribution patterns within the genotypes. The highest contents of anthocyanins were found in the middlings of black barley genotypes or in the shorts of blue and purple wheat. These data suggest the possibility to improve the antioxidant release from cereal-based food through selection of postharvest treatments.     

Phenolic Content and Antioxidant Activity of Pearled Wheat and Roller‐Milled Fractions

Wheat contains phenolic compounds concentrated mainly in bran tissues. This study examined the distribution of phenolics and antioxidant activities in wheat fractions derived from pearling and roller milling. Debranning (pearling) of wheat before milling is becoming increasingly accepted by the milling industry as a means of improving wheat rollermilling performance, making it of interest to determine the concentration of ferulic acid at various degrees of pearling. Eight cultivar samples were used, including five genotypes representing four commercial Canadian wheat classes with different intrinsic qualities. Wheat was pearled incrementally to obtain five fractions, each representing an amount of product equivalent to 5% of initial sample weight. Wheat was also roller milled without debranning. Total phenolic content of fractions was determined using the modified Folin‐Ciocalteau method for all pearling fractions, and for bran, shorts, bran flour, and first middlings flour from roller milling. Antioxidant activity was determined on phenolic extracts by a method involving the use of the free radical 2,2‐diphenyl‐l‐picrylhydrazyl (DPPH). Total phenolics were concentrated in fractions from the first and second pearlings (>4,000 mg/kg). Wheat fractions from the third and fourth pearlings still contained high phenolic content (>3,000 mg/kg). A similar trend was observed in antioxidant activity of the milled fractions with ≈4,000 mg/kg in bran and shorts, ≈3,000 mg/kg in bran flour, and <1,000 mg/kg in first middlings flour. Total phenolic content and antioxidant activity were highly correlated (R² = 0.94). There were no significant differences between red and white wheat samples. A strong influence of environment (growing location) was indicated. Pearling represents an effective technique to obtain wheat bran fractions enriched in phenolics and antioxidants, thereby maximizing health benefits associated with wheat‐based products.     

Antioxidant capacity of oat (Avena sativa L.) extracts. 1. Inhibition of low-density lipoprotein oxidation and oxygen radical absorbance capacity

Milled oat groat pearlings, trichomes, flour, and bran were extracted with methanol and the fractions tested in vitro for antioxidant capacity against low-density lipoprotein (LDL) oxidation and R-phycoerythrin protein oxidation in the oxygen radical absorbance capacity (ORAC) assay. The oxidative reactions were generated by 2,2'-azobis(2-amidinopropane) HCl (AAPH) or Cu(2+) in the LDL assay and by AAPH or Cu(2+) + H(2)O(2) in the ORAC assay and calibrated against a Trolox standard to calculate Trolox equivalents (1 Trolox equivalent = 1 TE = activity of 1 micromol of Trolox). The antioxidant capacity of the oat fractions was generally consistent with a potency rank of pearlings (2.89-8.58 TE/g) > flour (1.00-3.54 TE/g) > trichome (1.74 TE/g) = bran (1.02-1.62 TE/g) in both LDL and ORAC assays regardless of the free radical generator employed. A portion of the oat antioxidant constituents may be heat labile as the greatest activity was found among non-steam-treated pearlings. The contribution of oat tocols from the fractions accounted for <5% of the measured antioxidant capacity. AAPH-initiated oxidation of LDL was inhibited by the oat fractions in a dose-dependent manner, although complete suppression was not achieved with the highest doses tested. In contrast, Cu(2+)-initiated oxidation of LDL stimulated peroxide formation with low oat concentrations but completely inhibited oxidation with higher doses. Thus, oats possess antioxidant capacity most of which is likely derived from polar phenolic compounds in the aleurone.     

Determination of free flavan-3-ol content in barley (Hordeum vulgare L.) air-classified flours: comparative study of HPLC-DAD/MS and spectrophotometric determinations

The determination of free flavan-3-ol compounds in barley flours (cv. Gotic) and two resulting milling fractions (fine fraction 57% and coarse fraction 43%, w/w) obtained by air classification of dehulled grain is herein described. The determinations were carried out using reversed phase high-performance liquid chromatography coupled with both diode array detection and ESI-MS compared with conventional spectrophotometric determinations (total phenolic compounds by the Folin-Ciocalteu method and free radical scavenging activity with the DPPH assay). Significant correlations among the HPLC quantification, the spectrophotometric data, and the antioxidant capacity of extracts were revealed by Pearson's analysis. Nine flavan-3-ols were identified by HPLC-MS. Catechins and their derivatives were found to make a substantial contribution to the antioxidant power of extracts. The coarse fraction showed larger concentrations of flavan-3-ols (221%) with respect to the fine fraction. This was confirmed by the antioxidant activity of the analyzed flours. The coarse fraction showed the greatest antioxidant activity (1200.1 +/- 66.2 micromol of Trolox equiv/100 g of flour) with respect to whole meal and fine fraction (1025.9 +/- 18.3 and 761.7 +/- 55.3 micromol of Trolox equiv/100 g of flour, respectively).     

Antioxidant capacity of different broccoli (Brassica oleracea) genotypes using the oxygen radical absorbance capacity (ORAC) assay

Antioxidant capacity of hydrophilic and lipophilic extracts from eight broccoli genotypes was compared using the oxygen radical absorbance capacity (ORAC) assay. Each genotype was analyzed for carotenoid, tocopherol, ascorbic acid, and flavonoid content. Results indicate that the antioxidant capacity of hydrophilic extracts ranged from 65.8 to 121.6 micromol trolox equivalents (TE)/g of tissue, and the capacity of lipophilic extracts ranged from 3.9 to 17.5 micromol TE/g. Ascorbic acid and flavonoid content of the hydrophilic extracts did not explain the total variation in antioxidant capacity of those extracts, suggesting either the presence of other antioxidant components that have yet to be identified or that the known antioxidants are producing synergistic effects. The carotenoids did correlate with antioxidant capacity of the lipophilic extracts and accounted for the majority of the variability in that fraction. The variability in hydrophilic and lipophilic antioxidant capacity found among these genotypes suggests that potential efficacy from antioxidants will vary considerably from genotype to genotype.     

Composition and Distribution of Pentosans in Millstreams of Different Hard Spring Wheats

Six commercially grown samples of hard spring wheat were milled using a tandem Buhler laboratory mill. Individual flour streams and branny by‐products, as well as whole‐grain wheat and straight‐grade flour, were characterized in terms of total (TP), water‐extractable (WEP), and water‐unextractable (WUP) pentosans. One representative cultivar sample was analyzed for its ratio of arabinose to xylose (A/X). TP and WEP of whole grain wheat of the six samples had ranges of 5.45–7.32% and 0.62–0.90% (dm), respectively. Neither TP nor WEP of whole grain was related to ash content variation. There was significant variation in the distribution and composition of pentosans in 16 millstreams of all the wheat samples, including bran and shorts fractions; TP and WEP contents had ranges of 1.69–32.4% and 0.42–1.76% (dm), respectively. When ash contents exceeded ≈0.6% (dm), strong positive correlations were obtained between ash and TP contents, and between ash and WUP contents for all the millstreams. Among bran and shorts fractions, TP and WUP content increased in the order of coarse bran > fine bran > shorts; while WEP, WEP/WUP and A/X showed the opposite pattern of variation of shorts > fine bran > coarse bran. Bran and shorts fractions had pentosan contents several times higher than would be predicted from the relationship between pentosan and ash contents of the flour streams. Pentosans therefore represented a much more sensitive marker of flour refinement compared with ash content. Pentosans of endosperm were substantially different in their extractability and composition from those of bran. On this basis, different functionalities of pentosans of bran and endosperm would be expected. Results demonstrated the importance of milling extraction and millstream blending in the functionality and quality of wheat flour for breadmaking.     

Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity

Processed fruits and vegetables have been long considered to have lower nutritional value than their fresh commodities due to the loss of vitamin C during processing. This research group found vitamin C in apples contributed < 0.4% of total antioxidant activity, indicating most of the activity comes from the natural combination of phytochemicals. This suggests that processed fruits and vegetables may retain their antioxidant activity despite the loss of vitamin C. Here it is shown that thermal processing elevated total antioxidant activity and bioaccessible lycopene content in tomatoes and produced no significant changes in the total phenolics and total flavonoids content, although loss of vitamin C was observed. The raw tomato had 0.76 +/- 0.03 micromol of vitamin C/g of tomato. After 2, 15, and 30 min of heating at 88 degrees C, the vitamin C content significantly dropped to 0.68 +/- 0.02, 0.64 +/- 0.01, and 0.54 +/- 0.02 micromol of vitamin C/g of tomato, respectively (p < 0.01). The raw tomato had 2.01 +/- 0.04 mg of trans-lycopene/g of tomato. After 2, 15, and 30 min of heating at 88 degrees C, the trans-lycopene content had increased to 3.11+/- 0.04, 5.45 +/- 0.02, and 5.32 +/- 0.05 mg of trans-lycopene/g of tomato (p < 0.01). The antioxidant activity of raw tomatoes was 4.13 +/- 0.36 micromol of vitamin C equiv/g of tomato. With heat treatment at 88 degrees C for 2, 15, and 30 min, the total antioxidant activity significantly increased to 5.29 +/- 0.26, 5.53 +/- 0.24, and 6.70 +/- 0.25 micromol of vitamin C equiv/g of tomato, respectively (p < 0.01). There were no significant changes in either total phenolics or total flavonoids. These findings indicate thermal processing enhanced the nutritional value of tomatoes by increasing the bioaccessible lycopene content and total antioxidant activity and are against the notion that processed fruits and vegetables have lower nutritional value than fresh produce. This information may have a significant impact on consumers' food selection by increasing their consumption of fruits and vegetables to reduce the risks of chronic diseases.     

Insight into the distribution of arabinoxylans, endoxylanases, and endoxylanase inhibitors in industrial wheat roller mill streams

To gain insight into the distribution of arabinoxylans (AX), endoxylanases, and endoxylanase inhibitors in industrial wheat roller milling, all streams, that is, 54 flour fractions, 4 bran fractions, and the germ, were analyzed for ash, starch, and protein contents, alpha-amylase activity levels, total (TOT-AX) and water-extractable arabinoxylan (WE-AX) contents, endoxylanase activity levels, and endoxylanase inhibitor (TAXI and XIP) contents. In general, bran fractions were significantly richer in TOT-AX and WE-AX contents, endoxylanase activity levels, and endoxylanase inhibitor contents than germ and, even more so, than flour fractions. In the 54 different flour fractions, minimal and maximal values for TOT-AX and WE-AX contents differed by ca. 2-fold, whereas they differed by ca. 15-fold for endoxylanase activity levels. The latter were positively correlated with ash and negatively correlated with starch content, suggesting that the endoxylanase activity in flour is strongly influenced by the level of bran contamination. TAXI contents in the flour fractions varied ca. 4-fold and were strongly correlated with bran-related parameters such as ash content and enzyme activity levels, whereas XIP contents varied ca. 3-fold and were not correlated with any of the parameters measured in this study. The results can be valuable in blending and optimizing wheat flour fractions to obtain flours with specific technological and nutritional benefits.     

Decorticating sorghum to concentrate healthy phytochemicals

The growing prominence of nutrition-related health problems demands strategies that explore nontraditional natural ingredients to expand healthy food alternatives. Specialty sorghums were decorticated using a tangential abrasive dehulling device (TADD) to remove successive bran layers, which were collected at 1 min intervals and analyzed for phenols, tannins, 3-deoxyanthocyanins, dietary fiber, and antioxidant activity. The first two bran fractions had the highest levels of phenols and antioxidant activity (3-6 times as compared to whole grain). Brown (tannin-containing) and black sorghums had at least 10 times higher antioxidant activity than white sorghum or red wheat brans. Black sorghums had the highest 3-deoxyanthocyanin content (up to 19 mg/g bran). Dietary fiber in sorghum brans ranged between 36 and 45%, as compared to 48% for wheat bran. Specialty sorghum brans are rich in valuable dietary components and present promising opportunities for improving health attributes of food.     

Importance of insoluble-bound phenolics to antioxidant properties of wheat

Two commercial samples of soft (70% Canadian Eastern soft red spring and 30% Canadian Eastern soft white winter) and hard (90% Canadian western hard red spring and 10% Canadian Eastern hard red winter) wheats were used to obtain different milling fractions. Phenolics extracted belonged to free, soluble esters and insoluble-bound fractions. Soluble esters of phenolics and insoluble-bound phenolics were extracted into diethyl ether after alkaline hydrolysis of samples. The content of phenolics was determined using Folin-Ciocalteu's reagent and expressed as ferulic acid equivalents (FAE). The antioxidant activity of phenolic fractions was evaluated using Trolox equivalent antioxidant capacity, 2,2-diphenyl-1-picrylhydrazyl radical scavenging, reducing power, oxygen radical absorbance capacity, inhibition of oxidation of human low-density lipoprotein cholesterol and DNA, Rancimat, inhibition of photochemilumenescence, and iron(II) chelation activity. The bound phenolic content in the bran fraction was 11.3 +/- 0.13 and 12.2 +/- 0.15 mg FAE/g defatted material for hard and soft wheats, respectively. The corresponding values for flour were 0.33 +/- 0.01 and 0.46 +/- 0.02 mg FAE/g defatted sample. The bound phenolic content of hard and soft whole wheats was 2.1 (+/-0.004 or +/-0.005) mg FAE/g defatted material. The free phenolic content ranged from 0.14 +/- 0.004 to 0.98 +/- 0.05 mg FAE/g defatted milling fractions of hard and soft wheats examined. The contribution of bound phenolics to the total phenolic content was significantly higher than that of free and esterified fractions. In wheat, phenolic compounds were concentrated mainly in the bran tissues. In the numerous in vitro antioxidant assays carried out, the bound phenolic fraction demonstrated a significantly higher antioxidant capacity than free and esterified phenolics. Thus, inclusion of bound phenolics in studies related to quantification and antioxidant activity evaluation of grains and cereals is essential.     

Antioxidant activity of alkylresorcinols from rye bran and their protective effects on cell viability of PC-12 AC cells

Alkylresorcinols (ARs) are phenolic lipids that are present in high amounts in the bran layer of different cereals. Rye samples, cultivar Hazlet, and a white rye genotype, RT202, were analyzed for their antioxidant properties and AR content and composition, based on six fractions of the bran, where 1 was the outermost fraction and 6 was the bran fraction closest to the endosperm. Gas chromatography-mass spectrometry (GC-MS) analysis demonstrated that the most commonly found AR homologue in Hazlet rye is C19:0 and that the total amount of ARs decreases from the outermost to innermost fractions. The antioxidant activity using oxygen radical absorbance capacity (ORAC) for both white rye genotype RT202 and Hazlet brans was determined to decrease from the outermost fraction (136.05 μmol TE/g for Hazlet fraction 1 and 186.57 μmol TE/g for white rye genotype RT202 fraction 1) to the innermost fraction (9.84 μmol TE/g for Hazlet fraction 6 and 78.75 μmol TE/g for white rye genotype RT202 fraction 2). A positive relationship was seen with GC-MS results. Treatment of PC-12 AC cells with Hazlet fraction 1 increased mitochondrial biogenesis as determined using mitochondrial fluorescent dyes. In the presence of a prooxidant (AAPH), PC-12 AC cells were better protected from free radical attack when treated with Hazlet fraction 1 than with all other bran fractions. The results suggest that higher AR content in bran fractions confers antioxidant protection against free radical damage.     

Carotenoid, tocopherol, phenolic acid, and antioxidant properties of Maryland-grown soft wheat

Consumers' desires to either reduce the risk of or manage a specific health condition through improved diet have stimulated the research of agricultural products for their potential health beneficial components such as tocopherols and natural antioxidants. Soft wheat is one of the major crops in Maryland, with little information available about its potentially beneficial components. This study examined eight selected Maryland-grown soft wheat varieties or experimental lines for their potential beneficial components including tocopherols, carotenoids, total phenolics and phenolic acids and their antioxidant properties, including Fe(2+) chelating capacity and free radical scavenging activities against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH(*) ), radical cation ABTS(*)(+), and oxygen radical (ORAC). The results showed that all tested soft wheat grain samples contained alpha-tocopherol, with a range of 3.4-10.1 microg/g. Lutein was the primary carotenoid present in the grain samples at a level of 0.82-1.14 microg/g, along with significant amounts of zeaxanthin and beta-carotene. Vanillic, syringic, p-coumaric, and ferulic acids were found in soluble free, soluble conjugated, and insoluble bound forms in the grain extracts, with ferulic acid as the predominant phenolic acid. The eight soft wheat varieties differed in their antioxidant properties. The tested wheat grain samples exhibited ED(50) values against DPPH(*) of 23-27 mg of grain equiv/mL, ORAC of 32.9-48 micromol of Trolox equiv (TE)/g, and ABTS(*)(+) scavenging capacity of 14.3-17.6 micromol of TE/g. These data suggest the possibility of producing soft wheat varieties rich in selected health beneficial factors for optimum human nutrition though breeding programs.