Steryl phenolic acid esters in cereals and their milling fractions

The steryl ferulate contents of rye and wheat grains and their milling fractions were analyzed using a reversed-phase high-performance liquid chromatographic (HPLC) method. HPLC-mass spectrometry was used for identification. In addition, steryl ferulates of some selected milling byproducts were determined. The total steryl ferulate contents of rye and wheat grains were 6.0 and 6.3 mg/100 g, respectively. Uneven distribution of steryl ferulates in the grains led to considerable differences in the milling products; their steryl ferulate contents ranged from trace amounts in flours with low ash content to 20 and 34 mg/100 g in rye and wheat brans, respectively. Campestanyl ferulate and sitostanyl ferulate were the main components, followed by campesteryl ferulate and sitosteryl ferulate, whereas sitosterol was the main component in total sterols. Among the other samples, a byproduct of rice milling (pearling dust) was the best source of steryl ferulates, its total steryl ferulate content being 119 mg/100 g, whereas no measurable amounts of steryl ferulates were measured in oat bran or pearling dust of barley. The results indicated that rye and wheat and especially their bran fractions are comparable to corn as steryl ferulate sources.     

Content of gamma-oryzanol and composition of steryl ferulates in brown rice (Oryza sativa L.) of European origin

The content of gamma-oryzanol and the composition of steryl ferulates were determined in brown rice of European origin using on-line coupled liquid chromatography-gas chromatography (LC-GC). Analysis of 30 brown rice samples of various cultivars, grown at different sites and in different seasons, revealed the gamma-oryzanol content to range from 26 to 63 mg/100 g. Cycloartenyl ferulate and 24-methylenecycloartanyl ferulate were the major components of gamma-oryzanol followed by campesteryl ferulate, campestanyl ferulate, and beta-sitosteryl ferulate. The proportions of individual steryl ferulates exhibited enormous variability. However, irrespectively of the great variations observed for single steryl ferulates, the proportions of the sum of 4,4'-dimethylsteryl ferulates (cycloartenyl ferulate, 24-methylenecycloartanyl ferulate) and the sum of 4-desmethylsteryl ferulates (campesteryl ferulate, campestanyl ferulate, and beta-sitosteryl ferulate) were rather constant. The significant natural variability observed for gamma-oryzanol content and composition of steryl ferulates were shown to be influenced by environmental conditions but not by the degree of maturity of rice grains.     

Antioxidant activity of steryl ferulate extracts from rye and wheat bran

Antioxidant activity of steryl ferulates from other sources than rice have not yet been studied much, despite the fact that rice steryl ferulates (gamma-oryzanol) have been shown to possess good antioxidant activity. In this study, steryl ferulate extracts from wheat or rye bran were studied for their capability to inhibit hydroperoxide formation in bulk methyl linoleate and methyl linoleate emulsion. Further, their activity to scavenge DPPH radicals was analyzed. The activities were compared to synthetic steryl ferulates, rice steryl ferulates, ferulic acid, and alpha-tocopherol. Nonrice cereal extracts of steryl ferulates exhibited good antioxidant activity, especially in the bulk lipid system. The radical scavenging activity was similar to that of nonesterified ferulic acid, indicating that the ferulic acid moiety is responsible for the antioxidant properties. This study illustrates a new aspect to the health-promoting properties of rye and wheat.     

Effects of processing on availability of total plant sterols, steryl ferulates and steryl glycosides from wheat and rye bran

Rye and wheat bran are excellent natural sources of plant sterols in the diet. Their content, however, may vary according to processing. Thermal (roasting and heating in a microwave oven), mechanical (milling and cryogenic grinding), and enzymatic treatments (hydrolysis with xylanase or beta-glucanase or a mixture of these two enzymes) were performed, and their effect on sterol content, extractability of sterols and the characteristic steryl conjugates of cereals (steryl ferulates, steryl glycosides, and acylated steryl glycosides) were studied. Mechanical and enzymatic treatments increased the apparent sterol content, whereas aqueous processing without enzymes hindered the availability of total sterols, especially from rye bran. Changes were also seen in the amounts of steryl conjugates caused by the enzymatic treatments. On the basis of the results of this study, it can be speculated that a combination of fine particle size and enzymatic processing results in optimal sterol availability in cereal processing.     

Characterization of triterpene alcohol and sterol ferulates in rice bran using LC-MS/MS

Ferulic acid esters of triterpene alcohols and sterols in rice bran oil have been extensively studied and reported to possess important pharmacological actions. Inconsistent results on the numbers and structures of ferulates have been reported, primarily because of the analytical procedures employed. Conventional methods for analysis of phytosterol content in oil are carried out by characterization of trimethylsilylated derivatives (TMS) using GC-EI-MS after saponification of oils or individual compound isolated from oils. This study developed an LC-MS/MS method for the direct analysis of triterpene alcohol and sterol esters in rice bran oil. In addition to verifying the results of previous research, nine new relatively polar triterpene alcohol and sterol esters were characterized by their retention behaviors in LC and ESI-MS data from both negative- and positive-ion mode. This is the first evidence for the presence of hydroxylated ferulate esters and caffeate esters as part of gamma-oryzanol in rice bran. The method enables rapid and direct on-line characterization of triterpene alcohol and sterol esters in oils. LC-MS/MS equipped with reverse-phase LC and ESI-MS should be well-suited for identification and quantification of the polar metabolites of phytosterols in biological fluids after consumption of rice bran oil or other oils.     

Plant Sterols in Cereals and Cereal Products

The total plant sterol contents (free sterols and covalently bound structures) of the main cereals cultivated in Finland were determined. Furthermore, sterol contents were determined for different flour and bran fractions in the milling process of wheat and rye, as well as plant sterol contents in various milling and retail bakery products. The sample preparation procedure included acid and alkaline hydrolysis to liberate sterols from their glycosides and esters, respectively. Free sterols were extracted and, after recovery using solid‐phase extraction, derivatized to trimethylsilyl ethers for gas chromatography (GC) analysis. We used GC with a mass spectrometer (MS) for identification. When two cultivars of rye, wheat, barley, and oats grown in the same year were compared, the highest plant sterol content was observed in rye (mean content 95.5 mg/100 g, wb), whereas the total sterol contents (mg/100 g, wb) of wheat, barley, and oats were 69.0, 76.1, and 44.7, respectively. In addition, the 10 rye cultivars and breeding lines compared had total sterol contents of 70.7–85.6 mg/100 g. In the milling process of rye and wheat, the plant sterols fractionated according to the ash content of the corresponding milling product. In all cereal grain and milling product samples, sitosterol was the main sterol. The level of stanols differed in the different milling process samples; it was lower in the most refined rye and wheat flours (≈15%) than in the bran fractions (≈30% in the bran with 4% ash content). Rye bread with whole meal rye flour as the main or only ingredient was a good source of sterols. Sterol content was higher than that of wheat bread, whereas plant sterol content of other bakery products was affected by the type and amount of fat used in baking.     

Cross-linking of maize walls by ferulate dimerization and incorporation into lignin

Cross-linking of xylans and lignin by ferulates was investigated with primary maize walls acylated with 2% ferulate and with ferulate ethyl esters. Peroxidase-mediated coupling of wall ferulate and ethyl ferulate yielded mostly 8-coupled products, including three new dehydrodimers. Significant quantities of 5-5-coupled diferulate formed only within walls, suggesting that matrix effects influence dimer formation. Over 60% of wall ferulate dimerized upon H2O2 addition, suggesting that xylan feruloylation is highly regulated during wall biosynthesis to permit extensive dimer formation at the onset of lignification. During lignification, ferulate and 5-5-coupled diferulate copolymerized more rapidly and formed fewer ether-linked structures with coniferyl alcohol than 8-5-, 8-O-4-, and 8-8-coupled diferulates. The potential incorporation of most ferulates and diferulates into lignin exceeded 90%. As a result, xylans become extensively cross-linked by ferulate dimerization and incorporation to lignin, but only a small and variable proportion of these cross-links is measurable by solvolysis of lignified walls.     

Sterol ferulates, sterols, and 5-alk(en)ylresorcinols from wheat, rye, and corn bran oils and their inhibitory effects on Epstein-Barr virus activation

Sterol ferulate, free sterol, and 5-alk(en)ylresorcinol constituents of wheat, rye, and corn bran oils were studied. Among the sterol ferulates, one novel compound, 24-methylenecholestanol ferulate (7), along with six known compounds, namely, 24-methylcholestanol ferulate (1), 24-methylcholesterol ferulate (2), 2-methyllathosterol ferulate (3), stigmastanol ferulate (4), sitosterol ferulate (5), and schottenol ferulate (6), were isolated and characterized. Five known free sterols, namely, 24-methylcholesterol (8), stigmastanol (9), sitosterol (10), schottenol (11), and stigmasterol (12), were isolated and identified. 5-Alk(en)ylresorcinols were found in wheat and rye bran oils but not in corn bran oil. Of these, one new compound, 5-n-(2'-oxo-14'-Z-heneicosenyl) resorcinol (19), and seven known compounds, namely, 5-n-heptadecyl- (13), 5-n-nonadecyl- (14), 5-n-heneicosyl- (15), 5-n-tricosyl- (16), 5-n-pentacosyl- (17), 5-n-(14'-Z-nonadecenyl)- (18), and 5-n-(2'-oxoheneicosyl)resorcinols (20), were isolated and characterized. These compounds were evaluated with respect to their inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells, which is known to be a primary screening test for antitumor promoters. Four compounds, 1, 2, 4, and 11, showed potent inhibitory effects on EBV-EA induction.     

Triterpene alcohol and sterol ferulates from rice bran and their anti-inflammatory effects

Six novel feruloyl esters of triterpene alcohols and sterols, viz., two trans-ferulates, cycloeucalenol and 24-methylenecholesterol trans-ferulates, and four cis-ferulates, cycloartenol, 24-methyelenecycloartanol, 24-methylcholesterol, and sitosterol cis-ferulates, besides five known trans-ferulates, cycloartenol (CAR), 24-methylenecycloartanol (24-MCA), 24-methylcholesterol, sitosterol, and stigmastanol trans-ferulates, and one known cis-ferulate, stigmastanol cis-ferulate, were isolated from the methanol extract of edible rice bran. These and eight other synthetic trans- and cis-ferulates of triterpene alcohols and sterols, along with the corresponding free alcohols, were evaluated with respect to their anti-inflammatory activity against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1 microg per ear) in mice. All of the ferulates showed marked inhibitory activity, and their 50% inhibitory dose (ID(50)) was 0. 1-0.8 mg per ear. On the other hand, whereas two free triterpene alcohols, CAR and 24-MCA, showed strong inhibition (ID(50) 0.2-0.3 mg/ear), eight free sterols examined showed weaker activity (ID(50) 0.7-2.7 mg/ear) than their corresponding ferulates.     

Moderate ferulate and diferulate levels do not impede maize cell wall degradation by human intestinal microbiota

The degradation of plant fiber by human gut microbiota could be restricted by xylan substitution and cross-linking by ferulate and diferulates, for example, by hindering the association of enzymes such as xylanases with their substrates. To test the influence of feruloylation on cell wall degradability by human intestinal microbiota, nonlignified primary cell walls from maize cell suspensions, containing various degrees of ferulate substitution and diferulate cross-linking, were incubated in nylon bags in vitro with human fecal microbiota. Degradation rates were determined gravimetrically, and the cell walls were analyzed for carbohydrates, ferulate monomers, dehydrodiferulates, dehydrotriferulates, and other minor phenolic constituents. Shifting cell wall concentrations of total ferulates from 1.5 to 15.8 mg/g and those of diferulates from 0.8 to 2.6 mg/g did not alter the release of carbohydrates or the overall degradation of cell walls. After 24 h of fermentation, the degradation of xylans and pectins exceeded 90%, whereas cellulose remained undegraded. The results indicate that low to moderate levels of ferulates and diferulates do not interfere with hydrolysis of nonlignified cell walls by human gut microbiota.     

Carotenoid bioaccessibility from whole grain and degermed maize meal products

Although yellow maize (Zea mays) fractions and products are a source of dietary carotenoids, only limited information is available on the bioavailability of these pigments from maize-based foods. To better understand the distribution and bioavailability of carotenoid pigments from yellow maize (Z. mays) products, commercial milled maize fractions were screened for carotenoid content as were model foods including extruded puff, bread, and wet cooked porridge. Carotenoid content of maize fractions ranged from a low of 1.77-6.50 mg/kg in yellow maize bran (YCB) to 12.04-17.94 mg/kg in yellow corn meal (YCM). Lutein and zeaxanthin were major carotenoid species in maize milled fractions, accounting for approximately 70% of total carotenoid content. Following screening, carotenoid bioaccessibility was assessed from model foods using a simulated three-stage in vitro digestion process designed to measure transfer of carotenoids from the food matrix to bile salt lipid micelles (micellarization). Micellarization efficiency of xanthophylls was similar from YCM extruded puff and bread (63 and 69%), but lower from YCM porridge (48%). Xanthophyll micellarization from whole yellow corn meal (WYCM) products was highest in bread (85%) and similar in extruded puff and porridge (46 and 47%). For extruded puffs and breads, beta-carotene micellarization was 10-23%, but higher in porridge (40-63%), indicating that wet cooking may positively influence bioaccessibility of apolar carotenes. The results suggest that maize-based food products are good dietary sources of bioaccessible carotenoids and that specific food preparation methods may influence the relative bioaccessibility of individual carotenoid species.     

Bioavailability of inorganic arsenic in cooked rice: practical aspects for human health risk assessments

Arsenic is present in rice grain mainly as inorganic arsenic. Little is known about the effect of cooking on inorganic arsenic content in rice and its bioavailability. This study evaluated total arsenic and inorganic arsenic in rice cooked with arsenic-contaminated water, the bioaccessibility of As(III) and As(V) after simulated gastrointestinal digestion, and the extent of arsenic retention and transport by Caco-2 cells used as a model of intestinal epithelia. After cooking, inorganic arsenic contents increase significantly. After simulated gastrointestinal digestion, the bioaccessibility of inorganic arsenic reached 63-99%; As(V) was the main species found. In Caco-2 cells, arsenic retention, transport, and total uptake (retention + transport) varied between 0.6 and 6.4, 3.3 and 11.4, and 3.9 and 17.8%, respectively. These results show that in arsenic endemic areas with subsistence rice diets, the contribution of inorganic arsenic from cooked rice should be considered in assessments of arsenic health risk.     

Antioxidant activity of tocopherols, tocotrienols, and gamma-oryzanol components from rice bran against cholesterol oxidation accelerated by 2,2'-azobis(2-methylpropionamidine) dihydrochloride

The antioxidant activities of vitamin E (alpha-tocopherol, alpha-tocotrienol, gamma-tocopherol, and gamma-tocotrienol) and gamma-oryzanol components (cycloartenyl ferulate, 24-methylenecycloartanyl ferulate, and campesteryl ferulate) purified from rice bran were investigated in a cholesterol oxidation system accelerated by 2,2'-azobis(2-methylpropionamidine) dihydrochloride. All components exhibited significant antioxidant activity in the inhibition of cholesterol oxidation. The highest antioxidant activity was found for 24-methylenecycloartanyl ferulate, and all three gamma-oryzanol components had activities higher than that of any of the four vitamin E components. Because the quantity of gamma-oryzanol is up to 10 times higher than that of vitamin E in rice bran, gamma-oryzanol may be a more important antioxidant of rice bran in the reduction of cholesterol oxidation than vitamin E, which has been considered to be the major antioxidant in rice bran. The antioxidant function of these components against cholesterol oxidation may contribute to the potential hypocholesterolemic property of rice bran.     

Lignins and ferulate-coniferyl alcohol cross-coupling products in cereal grains

Plant cell walls containing suberin or lignin in the human diet are conjectured to protect against colon cancer. To confirm the existence of authentic lignin in cereal grain dietary fibers, the DFRC (derivatization followed by reductive cleavage) method was applied to different cereal grain dietary fibers. By cleavage of diagnostic arylglycerol-beta-aryl (beta-O-4) ether linkages and identification of the liberated monolignols, it was ascertained that lignins are truly present in cereal grains. From the ratios of the liberated monolignols coniferyl alcohol and sinapyl alcohol, it is suggested that lignin compositions vary among cereals. Furthermore, dimeric cross-coupling products, comprising ferulate and coniferyl alcohol, were identified in most cereal fibers investigated. These ferulate 4-O-beta- and 8-beta-coniferyl alcohol cross-coupled structures indicate radical cross-coupling of polysaccharides to lignin precursors via ferulate.     

Lipophilic extracts from banana fruit residues: a source of valuable phytosterols

The chemical composition of the lipophilic extracts of unripe pulp and peel of banana fruit 'Dwarf Cavendish' was studied by gas chromatography-mass spectrometry. Fatty acids, sterols, and steryl esters are the major families of lipophilic components present in banana tissues, followed by diacylglycerols, steryl glucosides, long chain fatty alcohols, and aromatic compounds. Fatty acids are more abundant in the banana pulp (29-90% of the total amount of lipophilic extract), with linoleic, linolenic, and oleic acids as the major compounds of this family. In banana peel, sterols represent about 49-71% of the lipophilic extract with two triterpenic ketones (31-norcyclolaudenone and cycloeucalenone) as the major components. The detection of high amounts of steryl esters (469-24405 mg/kg) and diacylglycerols (119-878 mg/kg), mainly present in the banana peel extract, explains the increase in the abundance of fatty acids and sterols after alkaline hydrolysis. Several steryl glucosides were also found in significative amounts (273-888 mg/kg), particularly in banana pulp (888 mg/kg). The high content of sterols (and their derivatives) in the 'Dwarf Cavendish' fruit can open new strategies for the valorization of the banana residues as a potential source of high-value phytochemicals with nutraceutical and functional food additive applications.     

Protease Treatment for the Stabilization of Rice Bran: Effects on Lipase Activity,Antioxidants, and Lipid Stability

Rice bran lipid is rapidly made rancid by endogenous lipase enzymes. To inactivate rice bran lipase, an enzymatic hydrolytic method was developed and then compared with the thermal method. The efficiency of five proteolytic enzymes including trypsin, chymotrypsin, papain, bromelain, and Flavourzyme enzyme to stabilize rice bran was investigated. Moreover, the antioxidant content and storage stability of enzymatically stabilized rice bran (ESRB), thermally stabilized rice bran (TSRB), and raw rice bran (RRB) were studied. Trypsin, chymotrypsin, and papain showed a higher rate of hydrolysis than bromelain and Flavourzyme enzyme. After 120 min of hydrolysis, rice bran hydrolyzed by trypsin, chymotrypsin, and papain inactivated 80, 86, and 79% of lipase activity, respectively, whereas lipase activity of rice bran hydrolyzed by bromelain and Flavourzyme enzyme was higher than the initial rice bran. With a similar lipase inactivation level, the cheaper papain was used to produce ESRB. Total phenolics content of ESRB was 52.89 and 94.10% higher than in RRB and TSRB, respectively. In addition, γ‐oryzanol content in ESRB was 2.23‐ and 2.05‐fold of that in RRB and TSRB, respectively. Lipase activity of RRB increased throughout the two months of storage, whereas no change in lipase activity was observed in ESRB and TSRB. At the end of storage, free fatty acid contents of RRB, TSRB, and ESRB were 15.30, 4.67, and 3.92%, respectively. We propose enzymatic hydrolysis by papain for stabilization of rice bran with high antioxidant content and storage stability.     

Purification and identification of components of gamma-oryzanol in rice bran Oil.

High-purity gamma-oryzanol was obtained from crude rice bran oil using a normal-phase preparative scale HPLC. A reverse-phase HPLC method was used for separating the individual components of gamma-oryzanol present in rice bran oil. Ten fractions were isolated and collected using the reverse-phase HPLC method, and their structures were identified. Identification was accomplished using GC/MS with an electron impact mass spectrum after components were transformed into trimethylsilyl ether derivatives. The 10 components of gamma-oryzanol were identified as Delta(7)-stigmastenyl ferulate, stigmasteryl ferulate, cycloartenyl ferulate, 24-methylenecycloartanyl ferulate, Delta(7)-campestenyl ferulate, campesteryl ferulate, Delta(7)-sitostenyl ferulate, sitosteryl ferulate, compestanyl ferulate, and sitostanyl ferulate. Three of these, cycloartenyl ferulate, 24-methylenecycloartanyl ferulate, and campesteryl ferulate, were major components of gamma-oryzanol.     

水网地区水稻土的含硒量及根外施硒对糙米硒含量的影响

本文通过对嘉善县平原水稻土硒含量特征和该区水稻硒含量之间关系的调查研究表明：（１）嘉善县平原水稻土,水溶态硒含量低于一般水稻土水溶态硒的临界值（０．０１０μｇ／ｇ）,这与该区的土壤母质、地形等因素相关;（２）低硒水稻土了水稻对硒的吸收积累,因此该县出产的稻米硒含量亦普遍较低,平均含硒量低于粮食硒的正常含量范围（０．０４０～０．０７０μｇ／ｇ）;（３）水稻地上各部分含硒量存在差异,糠、秸杆中的含硒量     

Effects of Commercial Processing on Antioxidants in Rice Bran

Rice bran contains high amounts of beneficial antioxidants including tocopherols, tocotrienols, and oryzanols. Current rice milling technology produces rice bran from different layers of the kernel caryopsis. Under current practices, these layers are combined and then steam‐extruded to form a stabilized rice bran pellet that is storage‐safe prior to oil extraction. Each of these rice bran intermediates can vary in antioxidant content. The objective of this study was to investigate the changes in selected antioxidants in rice bran from both long‐ and medium‐grain rice during commercial milling and bran processing. Rice bran collected from various milling breaks of a commercial system had varying antioxidant levels. Bran collected after milling break 2 had the highest levels of tocopherol and tocotrienol. Oryzanol concentration was significantly higher in outer bran layers. Results also indicate that the long‐grain rice bran averaged ≈15% more antioxidants than the medium‐grain rice bran.     

Bioaccessibility of carotenoids and vitamin e from pasta: evaluation of an in vitro digestion model

The present investigation aimed to expand the knowledge of bioaccessibility of carotenoids, tocopherols, and tocotrienols from cereal products such as pasta. Because most of the published approaches assessing the bioaccessibility of lipophilic micronutrients dealt with fruits and vegetables, a prevalent in vitro digestion procedure was modified. Additionally, several digestion parameters were evaluated with regard to their impact on the bioaccessibility of carotenoids and vitamin E from pasta. Overall, the estimated values were highly dependent on the amount of bile extract present in the digestive medium and to a lesser extent on the simulated gastric pH and the incubation time with digestive enzymes. The bioaccessibility of carotenoids and vitamin E from durum wheat pasta was quite high (71 ± 5 and 70 ± 4%, respectively), whereas these micronutrients were considerably less accessible from pasta containing 10% eggs (57 ± 1 and 49 ± 5%, respectively).