The effect of in vitro digestion on steryl ferulates from rice (Oryza sativa L.) and other grains

Polished and cargo rice, wild rice, rice bran, corn bran, and wheat bran were subjected to a static in vitro digestion model, to monitor changes in their steryl ferulate content and composition. Free sterols, possible hydrolysis products of steryl ferulates, were also measured. Additionally, steryl ferulate bioaccessibility was calculated as the percentage of steryl ferulates liberated from the grain matrix into the digestive juice. Steryl ferulate content ranged between 6.1 and 3900 μg/g and decreased by 1-63% due to digestion. A parallel increase in free sterols of more than 70% was observed in all samples. Additionally, bioaccessibility of steryl ferulates was found to be almost negligible. These findings suggest that intestinal enzymes immediately hydrolyze steryl ferulates, which are liberated from the grain matrix, and thus they are practically unavailable for absorption in the small intestine. This further indicates that the hydrolysis products of steryl ferulates could be bioactive in the gut.     

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.     

The effect of calcium nutrition on the sterol content of tobacco roots and their susceptibility to Phytophthora parasitica var. nicotianae

The influence of nutrient solutions containing low (25 mg/1) and high (250 mg/1) calcium (Ca) concentrations on the total lipid, free sterol, sterol glycoside and steryl ester content and on the subsequent susceptibility of three flue‐cured tobacco cultivars to black shank, incited by Phytophthora parasitica var. nicotianae was investigated. The total lipid, total sterol and free sterol content differed appreciably among root systems of the three cultivars grown at high Ca. Roots of plants grown at low Ca contained relatively higher proportions of steryl glycosides and lower proportions of free sterols than roots of plants grown at high Ca, although the absolute sterol concentrations also differed significantly among the roots of cultivars grown at low Ca. Black shank disease development was significantly lower in roots of susceptible plants grown at low Ca than in roots of the same cultivar grown at high Ca. The innate susceptibility of the cultivars were not related to the lipid or sterol content. The reduced disease development of plants grown at low Ca appeared to be associated with changes in membrane permeability, which may be correlated with changes in the various sterol fractions.     

Cholesterol-lowering effects of plant steryl and stanyl laurate by oral administration in mice

The present study was conducted to investigate the efficacy of synthesized plant steryl and stanyl laurate in lowering the cholesterol level and to further examine the cholesterol-lowering potential of the free plant sterols and stanols dissolved in liquid emulsion on serum and liver lipids in mice by oral administration. Experimental results showed that both plant steryl and stanyl laurate could significantly decrease the serum levels of TC, LDL-C, LDL-C/HDL-C, and liver cholesterol contents and markedly increase fecal cholesterol concentrations but have no effect on serum TAG level, indicating that the produced plant steryl and stanyl laurate retained the cholesterol-lowering potential of natural plant sterols and stanols. However, no statistical difference in cholesterol-lowering efficacy was observed between plant steryl laurate and plant stanyl laurate, and free plant sterols and stanols dissolved in liquid emulsion could also significantly decrease serum cholesterol levels and markedly increase fecal cholesterol excretion. These results suggested that the esterified plant sterols/stanols had comparable effects to the free plant sterols/stanols in lowering serum TC levels but that they did gain a solubility advantage from the free plant sterols/stanols. Therefore, plant steryl/stanyl laurate could be considered as a potential nutraceutical or functional ingredient to reduce or prevent atherosclerosis and its related complications.     

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.     

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.     

Phytosterol composition of nuts and seeds commonly consumed in the United States

Phytosterols were quantified in nuts and seeds commonly consumed in the United States. Total lipid extracts were subjected to acid hydrolysis and then alkaline saponfication, and free sterols were analyzed as trimethylsilyl derivatives by capillary GC-FID and GC-MS. Delta5-Avenasterol was quantified after alkaline saponification plus direct analysis of the glucoside. Sesame seed and wheat germ had the highest total phytosterol content (400-413 mg/100 g) and Brazil nuts the lowest (95 mg/100 g). Of the products typically consumed as snack foods, pistachio and sunflower kernel were richest in phytosterols (270-289 mg/100 g). beta-Sitosterol, Delta5-avenasterol, and campesterol were predominant. Campestanol ranged from 1.0 to 12.7 mg/100 g. Only 13 mg/100 g beta-sitosterol was found in pumpkin seed kernel, although total sterol content was high (265 mg/100 g). Phytosterol concentrations were greater than reported in existing food composition databases, probably due to the inclusion of steryl glycosides, which represent a significant portion of total sterols in nuts and seeds.     

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.     

Characterization by gas chromatography/mass spectrometry of sterols in saccharomyces cerevisiae during autolysis.

Yeast autolysis affects membrane stability and induces a release of vacuolar enzymes into the cell cytoplasm. Consecutively, it was important to study the evolution of sterol content in Saccharomycescerevisiae for a fourteen day period of accelerated autolysis. Unesterified and esterified sterols were analyzed both in the biomass and in the autolysis medium. Ten sterols were identified by gas chromatography/mass spectrometry. A second group of six sterols was separated and partially characterized. Among the first group of 10 sterols, a dehydroergosterol was identified as ergosta-5, 7,9(11),22-tetraen-3beta-ol, not yet charaterized in S. cerevisiae. Yeast autolysis induced a decrease of esterified sterol content, especially first intermediates in the sequence of the ergosterol biosynthesis, as zymosterol. In contrast, the yeast autolysis resulted in the release of a low quantity of sterols into the medium. At the end of the fourteenth day of autolysis, 0.015% of the total sterol content of the initial biomass was found in the medium.     

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.     

Physical and Milling Characteristics of Wheat Kernels After Enzyme and Acid Treatments

The current wheat milling process separates bran from endosperm by passing tempered wheat kernels through successive break rolls and sifters. Using hydrolytic enzymes during tempering degrades bran and aleurone layers and can improve milling efficiency and yield. This study was conducted to evaluate the effects of chemical and enzymatic treatments of wheat kernels before milling on physical and milling characteristics of the resulting wheat and flour quality. Hard wheat kernels were soaked in dilute acid or water and dried back to original moisture before being tempered with enzymes in water. Kernel physical and milling characteristics (600 g) were evaluated. Dilute acid soaking did not affect the 1,000‐kernel weight and diameter but softened treated kernels. When treated kernels were pearled, bran removal was mostly from ends; and the reducing sugar content in enzyme‐treated bran was significantly higher than the control. Compared with the control, acid‐soaked enzyme‐tempered kernels showed small but significant improvement in straight flour yield, with virtually no difference in protein content, and flour color. Chemical and enzyme treatment resulted in higher ash in flour. These differences were not seen in milling of larger batches (1,500 g) of kernels.     

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.     

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.     

Protein extraction from heat-stabilized defatted rice bran. 1. Physical processing and enzyme treatments

Physical processing with or without enzyme treatments on protein extraction from heat-stabilized defatted rice bran (HDRB) was evaluated. Freeze-thaw, sonication, high-speed blending, and high-pressure methods extracted 12%, 15%, 16%, and 11% protein, respectively. Sonication (0-100%, 750 W), followed by amylase and combined amylase and protease treatments, extracted 25.6-33.9% and 54.0-57.8% protein, respectively. Blending followed by amylase and protease treatment extracted 5.0% more protein than the nonblended enzymatic treatments. High-pressure treatments, 0-800 MPa, with water or amylase-protease combinations, extracted 10.5-11.1% or 61.8-66.6% protein, respectively. These results suggest that physical processing in combination with enzyme treatments can be effective in extracting protein from HDRB.     

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.     

Chemical composition of abaca (Musa textilis) leaf fibers used for manufacturing of high quality paper pulps

The chemical composition of leaf fibers of abaca (Musa textilis), which are commonly used for high-quality paper pulp production, was thoroughly studied. The results revealed that the lignin content was 13.2% of the total fiber. The analysis of abaca fibers by pyrolysis coupled to gas chromatography-mass spectrometry (Py-GC/MS) released predominantly compounds arising from lignin and p-hydroxycinnamic acids, with high amounts of 4-vinylphenol. The latter compound was demonstrated to arise from p-coumaric acid by pyrolysis of abaca fibers in the presence of tetramethylammonium hydroxide, which released high amounts of p-coumaric acid (as the methyl derivative). Products from p-hydroxyphenyl (H), guaiacyl (G), and syringyl (S) propanoid units, with a predominance of the latter (H:G:S molar ratio of 1.5:1:4.9), were also released after Py-GC/MS of abaca fibers. Sinapyl and coniferyl acetates, which are thought to be lignin monomer precursors, were also found in abaca. The extractives content of the abaca fiber (0.4%) was low, and the most predominant compounds were free sterols (24% of total extract) and fatty acids (24% of total extract). Additionally, significant amounts of steroid ketones (10%), triglycerides (6%), omega-hydroxyfatty acids (6%), monoglycerides (4%), fatty alcohols (4%), and a series of p-hydroxycinnamyl (p-coumaric and ferulic acids) esterified with long chain alcohols and omega-hydroxyfatty acids were also found, together with minor amounts of steroid hydrocarbons, diglycerides, alpha-hydroxyfatty acids, sterol esters, and sterol glycosides.     

Characterization of the chemical composition of lotus plumule oil

Characterizations of lotus plumule and plumule oil, focusing on approximate composition analysis of lotus plumule powder and fatty acid composition, lipid classes, triglyceride (TG) profiles, and sterol analysis of the plumule oil, were conducted in this work. The results revealed that the lotus plumule constitutes 7.8% moisture, 4.2% ash, and 12.5% crude oil and 26.3% protein on the dry base. Lotus plumule oil is rich in linoleic acid (50.4%) and oleic acid (13.5%), and the dominating saturated fatty acids are palmitic acid (18.0%) and behenic acid (6.8%). The principal components of TG in lotus plumule oil are LLL (12.80%), beta-PLL (11.27%), beta-POL (8.28%), beta-PLO (8.58%), and beta-BeLL (8.32%). Lipid class assay of the crude oil gave the saponification value of 153.4 KOH mg/g and tocopherol content 390 mg/100 g. A distinct characteristic of lotus plumule oil is that its unsaponifiable matter is incredibly high, up to 14-19%, which consists mainly of beta-sitosterol (32%), Delta(5)-avenasterol (20%), and campesterol (6.3%). The major occurring form of sterols was found to be steryl ester. This work might be useful to develop innovative applications of lotus plumule oil.     

Extrusion Conditions Modify Hypocholesterolemic Properties of Wheat Bran Fed to Hamsters

Wheat bran was extruded in a twin‐screw extruder at five specific mechanical energy (SME) levels (0.120, 0.177, 0.234, 0.291, and 0.358 kWh/kg, dwb) and the cholesterol‐lowering effects were compared with those of unprocessed wheat bran when fed to four‐week‐old male golden Syrian hamsters (n = 10/treatment) for three weeks. Diets contained 10% total dietary fiber, 10.3% fat, 3% nitrogen, and 0.4% cholesterol. Plasma total cholesterol and very‐low‐density lipoprotein cholesterol were significantly lower with 0.120 kWh/kg extruded wheat bran diet compared with the unextruded wheat bran control. Total triglycerides were significantly lower with 0.120 and 0.177 kWh/kg wheat bran diets compared with those fed 0.291 and 0.358 kWh/kg extruded wheat bran diets. Cholesterol digestibility, total liver cholesterol, and total liver lipids were significantly lower with all the extruded wheat bran diets compared with the unextruded wheat bran control. Cholesterol digestibility for the 0.291 kWh/kg wheat bran diet was also significantly lower than all other extruded diets. Significantly more sterols were excreted with diets containing 0.291 and 0.358 kWh/kg extruded wheat bran compared with the unextruded wheat bran control. Wheat bran extruded with 0.291 kWh/kg diet resulted in a 13% reduction in plasma cholesterol and a 29% reduction in low‐density lipoprotein cholesterol. Considering lowest cholesterol digestibility, significantly higher sterol excretion, desirable plasma lipo‐protein cholesterol profile, significantly lower liver weight, total liver lipids, and liver cholesterol, the wheat bran extruded at 0.291 kWh/kg appeared to have the most desirable healthful potential. Data suggest that cholesterol‐lowering potential of wheat bran could be enhanced by optimizing the energy input used in the extrusion process.     

Effect of Extrusion on Hypocholesterolemic Properties of Rice,Oat, Corn,and Wheat Bran Diets in Hamsters

Brans from rice, oats, corn, and wheat were cooked in a twin-screw extruder at either high or low energy input, and their cholesterol-lowering effects were compared with those of unprocessed brans when fed to four-week-old male golden Syrian hamsters (n = 10 per treatment) for three weeks. Peanut oil was added to oat, corn, and wheat bran during the extrusion process to match the oil content of rice bran. Diets contained 10% total dietary fiber, 10.3% fat, 3% nitrogen, and 0.3% cholesterol. Plasma and liver cholesterol and total liver lipids were significantly lower with low-energy extruded wheat bran compared with unprocessed wheat bran. Extrusion did not alter the hypocholesterolemic effects of rice, oat, or corn brans. Plasma and liver cholesterol levels with corn bran were similar to those with oat bran. Relative cholesterol-lowering effects of the brans, determined with pooled (extruded and unextruded) bran data, were rice bran > oat bran > corn bran > wheat bran. Rice bran diets resulted in significantly lower levels of total plasma cholesterol and very low density lipoprotein cholesterol compared with all other brans. Total liver cholesterol and liver cholesterol concentrations (mg/g) were significantly lower with high-energy extruded rice bran compared with the cellulose control group. Plasma cholesterol and total liver cholesterol values with low-energy extruded wheat bran were similar to those with rice bran (unextruded or extruded) diets. Lowered cholesterol with rice bran diets may result in part from greater lipid and sterol excretion with these diets. Results with low-energy extruded wheat bran suggest that this type of processing may improve the potential for lowering cholesterol with wheat bran products.