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.     

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.     

挤压稳定化处理对米糠各组分蛋白结构及功能性质的影响

为了研究挤压稳定化处理对米糠各组分蛋白结构和功能性质的影响,选取龙粳31号大米米糠做为原料,采用双螺杆挤压技术对该原料进行稳定化处理。结果表明：米糠各组分蛋白在挤压处理后溶解性、起泡性和持油性显著降低（P?0.05）,持水性、起泡稳定性和乳化稳定性升高,谷蛋白持水性提高的幅度最大,较挤压前提高了39%。米糠谷蛋白的乳化活性与其他两种组分蛋白差异显著,清蛋白和球蛋白较挤压前分别降低5%和10%,谷蛋白乳化活性增加,较挤压前增加8%。结构特性分析结果表明产生这种差异的主要原因不是分子间作用力,而是挤压后各组分蛋白发生重组,形成大的聚集体过程中二级结构的变化截然相反,米糠清蛋白α-螺旋、β-转角和无规则卷曲含量都有所降低,β-折叠含量增势明显提高。挤压后的米糠谷蛋白结构与白蛋白显示出不同的趋势,谷蛋白的二级结构在酰胺I带变化显著,α-螺旋、β-转角与无规则卷曲的含量有所提高,β-折叠的含量下降。结果可为米糠各组分蛋白的工业化制备及在各种食品配方中的应用提供理论支撑。     

Aroma components of acid-hydrolyzed vegetable protein made by partial hydrolysis of rice bran protein

Hydrolyzed vegetable protein (HVP) was prepared from rice bran protein concentrate (RBPc) by partial hydrolysis with aqueous 0.5 N HCl at 95 degrees C for 12 or 36 h (H-RBPc-12 and H-RBPc-36, respectively). Aroma components of the RBPc and the HVPs were characterized by gas chromatography-olfactometry, gas chromatography-mass spectrometry, aroma extract dilution analysis, and calculation of odor activity values (OAVs). The predominant odorants in RBPc were 3-methylbutanal, hexanal, 2-aminoacetophenone, (E)-2-nonenal, phenylacetaldehyde, and beta-damascenone. Among these, the odor of 2-aminoacetophenone, present at 59 ng/g in RBPc, was reminiscent of the typical odor of RBPc. Most of the predominant odorants had higher log3FD factors in the H-RBPc-36 as compared to H-RBPc-12. Aroma impact compounds of H-RBPc-12 and H-RBPc-36 were 2-methoxyphenol (guaiacol), 4-hydroxy-2,5-dimethyl-3(2H)furanone, 3-hydroxy-4,5-dimethyl-2(5H)furanone (sotolon), vanillin, 3-methylbutanal, (E)-2-nonenal, 4-vinyl-2-methoxyphenol (p-vinylguaiacol), and beta-damascenone. Guaiacol had the highest OAV values of 2770 and 17650 in H-RBPc-12 and H-RBPc-36, respectively.     

Extraction of natural vitamin E from wheat germ by supercritical carbon dioxide

An efficient supercritical fluid extraction (SFE) process with carbon dioxide (SFE-CO(2)) was developed for the extraction of natural vitamin E (V(E)) from wheat germ. Both the pretreatment of extracted wheat germ and extraction conditions were optimized to ensure maximal V(E) yield. The extraction was undertaken at the extracting pressure of 4000-5000 psi, the extracting temperature of 40-45 degrees C, and the carbon dioxide flow rate of 2.0 mL/min for 90 min. An optimized pretreatment of wheat germ was usually necessary with a particle size of 30 mesh and a moisture content of 5.1%. A yield comparison of V(E) and its isomers extracted by supercritical CO(2) with those by conventional solvent extraction suggested that this SFE process was a practical process prospectively superior to conventional solvent extraction to prepare V(E) from wheat germ.     

亚临界丙烷萃取米糠油及其微量活性组分动力学模型

为了揭示亚临界流体萃取米糠油及其微量活性组分的动力学规律,该文以亚临界丙烷为萃取介质,研究了米糠油、γ-谷维素及α-生育酚等活性组分在不同萃取温度和时间的萃取率,并基于 Baümler 模型对目标物的萃取率进行模型拟合,建立相应的动力学模型并验证。结果表明,亚临界萃取过程中目标物的相对萃取率随时间增长递增,且在初始阶段（洗涤过程）的增长速率快于第二阶段（扩散过程）的增长速率;同时,升高萃取温度可同时提高目标产物的扩散系数和相对萃取率;基于Baümler模型可较好拟合亚临界丙烷萃取米糠油、γ-谷维素和α-生育酚的动力学过程（R2>0.95）,所得动力学模型具有较好的预测准确性。由Arrhenius方程可计算出米糠油、α-生育酚和γ-谷维素的亚临界萃取反应活化能,其中米糠油的活化能最低（5.23 kJ/mol）,α-生育酚次之（7.05 kJ/mol）,γ-谷维素最高（9.11 kJ/mol）,表明γ-谷维素最难萃取且对温度依赖程度最高;根据拟合所得动力学模型,对米糠油提取率进行预测,结果表明,该模型对米糠油萃取率的预测准确率达95.8%,而对γ-谷维素和α-生育酚萃取率的预测准确度稍低,分别达94.7%和94.4%,对三者均具有很好的预测准确性。总之,基于 Baümler 模型可以较好地建立米糠油及其有益伴随物的亚临界萃取动力学模型,从而较为准确的预测米糠油及其微量活性组分的萃取率,对米糠油的亚临界萃取工艺具有一定的理论指导意义。     

Influence of growth temperature on the amounts of tocopherols, tocotrienols, and gamma-oryzanol in brown rice

Brown rice is a valuable source of lipid-soluble antioxidants including ferulated phytosterols (i.e., gamma-oryzanol), tocopherols, and tocotrienols. To evaluate the impact of temperature on the accumulation of these compounds, seeds from six different rice lines grown to maturity in replicate greenhouses in Gainesville, FL, were analyzed. The lines represented Oryza sativa indica, O. sativa japonica, and Oryza glaberrima of different origins. Temperatures were maintained near ambient at one end of each greenhouse and at approximately 4.5 degrees C above ambient at the other end. gamma-Oryzanols, tocopherols, and tocotrienols were extracted from whole seed (i.e., brown rice) and analyzed by HPLC. Tocotrienols and tocopherols varied widely between lines but changed only slightly with respect to temperature. In general, the proportions of alpha-tocotrienol and/or alpha-tocopherol increased at elevated temperature, whereas gamma-tocopherol and gamma-tocotrienol decreased. Six gamma-oryzanol peaks, identified on the basis of absorbance maxima at 330 nm and HPLC-mass spectrometry, were quantified. The most abundant component was 24-methylenecycloartanyl ferulate, present at 40-62% of total. Its levels increased 35-57% at elevated temperature in five of six lines, accounting for most of the change in total gamma-oryzanol. The results suggest that the physiological action of individual ferulated phytosterols should be investigated because their relative proportions in gamma-oryzanol can change.     

Preparation and functional properties of rice bran protein isolate

Rice bran protein isolate (RBPI) containing approximately 92.0% protein was prepared from unstabilized and defatted rice bran using phytase and xylanase. The yield of RBPI increased from 34% to 74.6% through the use of the enzymatic treatment. Nitrogen solubilities of RBPI were 53, 8, 62, 78, 82, and 80% at pHs 2.0, 4.0, 6.0, 8.0, 10.0, and 12.0, respectively. Differential scanning calorimetry showed that RBPI had denaturation temperature of 83.4 degrees C with low endotherm (0.96 J/g of protein). RBPI had similar foaming properties in comparison to egg white. But emulsifying properties of RBPI were significantly lower than those of bovine serum albumin. The result of amino acid analysis showed that RBPI had a similar profile of essential amino acid requirements for 2-5-year-old children in comparison to that of casein and soy protein isolate.     

Prevention of hydrolytic rancidity in rice bran during storage.

The effect of microwave heating, packaging, and storage temperature on the production of free fatty acids (FFA) in rice bran was examined. Freshly milled raw rice bran was adjusted to 21% moisture content and heated in a microwave oven at 850 W for 3 min. Raw and microwave-heated rice bran were packed in zipper-top bags or vacuum-sealed bags and stored at 4-5 or 25 degrees C for 16 weeks. FFA content of bran was measured at 4-week intervals. Total FFA increased rapidly over the 16-week period from the initial value of 2.5% in raw bran stored at 25 degrees C to 54.9% in vacuum bags and 48.1% in zipper-top bags. However, total FFA of raw bran stored at 4-5 degrees C increased at a slower rate from an initial value of 2. 5 to 25.4% in vacuum bags and 19.5% in zipper-top bags. After 16 weeks of storage, total FFA of microwave-heated bran stored at 25 degrees C increased from 2.8 to 6.9 and 5.2%, respectively, for samples stored in vacuum bags and zipper-top bags. Total FFA of microwave-heated samples stored at 4-5 degrees C did not change significantly with storage time. Results showed that hydrolytic rancidity of rice bran can be prevented by microwave heating and that the recommended storage condition for microwaved rice bran is 4-5 degrees C in zipper-top bags.     

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.     

Prevention of oxidative rancidity in rice bran during storage.

The effect of microwave heat on lipoxygenase (LOX) activity in rice bran under various storage conditions was examined. Raw rice bran from the long-grain variety Lemont was adjusted to 21% moisture content and heated in a microwave oven at 850 W for 3 min. Raw and microwave-heated rice bran samples were packed in zipper-top bags or vacuum packs and stored at room temperature (25 degrees C) or in the refrigerator (4-5 degrees C) for 16 weeks. Samples were analyzed for LOX activity at 4-week intervals. LOX activity did not significantly change from its initial value at week 0 for zipper-top and vacuum-packed samples while stored at 4-5 degrees C for 12 weeks, but decreased at week 16. Vacuum packing did not show a significant impact on LOX activity during 16 weeks of storage. Microwave-heated samples stored in the refrigerator did not show significant change in LOX activity for up to 12 weeks but showed a significant (p < 0. 05) decrease at 16 weeks. Results showed that oxidative rancidity of rice bran could be prevented by microwave heating the samples, packing in zipper-top bags, and storing at 4-5 degrees C for up to 16 weeks.     

Use of microscopy to assess bran removal patterns in milled rice

During rice milling, the bran and germ are successively removed from the caryopsis (kernel). Because bran and germ contain large quantities of lipid, the amount of lipid remaining on the kernel surface may be used as a method for the assessment of milling quality. Bulk samples of rice pureline varieties and an experimental hybrid were milled for 0, 10, 20, 30, and 40 s. Scanning electron microscopy (SEM) revealed that brown rice kernels had large contours of linear protuberances and depressions running lengthwise along the kernel surface. The protuberances were abraded successively during milling, but varying amounts of material remained in the depressions. Light microscopy combined with the lipid-specific probes Nile Blue A or Sudan Black B demonstrated that the material in the depressions observed with SEM was lipid. Sections of whole, milled rice kernels, prepared using a modified sectioning technique and stained with Nile Blue A, showed that portions of the embryo remain after milling and that lipid is located on or near the surface of the kernel. Differences in quantity and distribution of residual lipid as milling duration increased were documented photographically to indicate the extent to which the bran and embryo components were removed during milling. This paper provides proof of concept that residual lipid is a robust measure of the degree of milling.     

Impact of browning reactions and bran pigments on color of parboiled rice

Rice color changes from white to amber during parboiling (soaking and steaming). Color parameters indicated that, during soaking, yellow bran pigments leached out in the water. The levels of the Maillard precursors (i.e., reducing sugars (RS) and free alpha-amino nitrogen (FAN)) depended on soaking temperature and time: leaching of RS was compensated by enzymic formation for long soaking times (>60 min), while proteolytic activity was too low to compensate for FAN leaching. Rice soaking under nitrogen, oxygen, or ambient conditions and determination of polyphenol oxidase activity allowed us to conclude that the effect of enzymic color changes on the soaked rice color was rather small. Color measurements of brown and milled mildly, intermediately, and severely parboiled rice samples showed that both brown and milled rice samples were darker and more red and yellow after parboiling and that the effect depended on the severity of parboiling conditions. Furthermore, steaming affected the rice color more and in a way opposite to that observed in soaking. The changes in RS and the loss of FAN during parboiling suggested that Maillard type reactions occur during brown rice steaming. Analyses of furosine levels confirmed Maillard browning of outer bran layers and endosperm during steaming. The level of this Maillard indicator increased with the severity of parboiling conditions in both brown and milled parboiled rice. Measurements of the levels of bran pigments indicated that bran pigments diffuse into the endosperm during parboiling and contribute to the parboiled rice color.     

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.     

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.     

Water extract of defatted rice bran suppresses visceral fat accumulation in rats

Rice bran has been reported to inhibit pancreatic lipase activity in vitro. This action shows that administration of rice bran may result in a decrease in plasma triglyceride levels and suppress accumulation of fat in vivo. We administered water extract of defatted rice bran (WED-rice bran) to rats to determine its effects. Single administration of WED-rice bran at a dose of 1 g/kg body weight caused a decrease in plasma triglyceride levels in fat emulsion induced hypertriglyceridemic rats. Four week administration of WED-rice bran suppressed accumulation of visceral fat and body weight gain without influencing food consumption, liver function, and renal function. These results indicate that a reduction of plasma triglycerides and suppression of visceral fat accumulation may be induced by pancreatic lipase inhibition caused by administration of WED-rice bran.     

Lipase-catalyzed modification of rice bran oil to incorporate capric acid

Capric acid (C10:0) was incorporated into rice bran oil with an immobilized lipase from Rhizomucor miehei as the biocatalyst. Effects of incubation time, substrate mole ratio, enzyme load, and water addition on mole percent incorporation of C10:0 were studied. Transesterification was performed in an organic solvent, hexane, and under solvent-free condition. Pancreatic lipase-catalyzed sn-2 positional analysis and tocopherol analysis were performed before and after enzymatic modification. Products were analyzed by gas-liquid chromatography (GLC) for fatty acid composition. After 24 h of incubation in hexane, there was an average of 26.5 +/- 1.8 mol % incorporation of C10:0 into rice bran oil. The solvent-free reaction produced an average of 24.5 +/- 3.7 mol % capric acid. In general, as the enzyme load, substrate mole ratio, and incubation time increased, the mole percent of capric acid incorporation also increased. Time course reaction indicated C10:0 incorporation increased up to 27.0 mol % at 72 h, for the reaction in hexane, and up to 29.6 mol % at 12 h, for the solvent-free reaction. The highest C10:0 incorporations (53.1 and 43.2 mol %) for the mole ratio experiment occurred at a mole ratio of 1:8 for solvent and solvent-free reactions, respectively. The highest C10:0 incorporation (27.9 mol %) for the reaction in hexane occurred at 10% enzyme load, and the highest incorporation (34.4 mol %) for the solvent-free reaction occurred at 20% enzyme load. Incorporation of C10:0 into rice bran oil declined with the addition of increasing amounts of water after reaching 30.3 mol % at 2% water addition in hexane, and in the solvent-free reaction after reaching 35.9 mol %.     

原料组分及应力松弛与方便米线品质的关系

以福昌、九峰、兴业、一枝秀等4种大米为原料制作方便米线,研究了米线品质特征与原料米化学成分、淀粉凝胶应力松弛特性的相关性,并结合应力松弛参数探讨了原料成分对米线品质的影响。结果表明,方便米线最大剪切应力为10.34~18.39 g/mm2,平均剪切应力为4.72~6.56 g/mm2,拉伸强度为7.77~15.72 g/mm2,拉伸形变为53.14~80.84 %,干物质吸水率为98.80~153.26 %,煮沸损失为12.09~24.39 %;原料大米粗蛋白含量越高,粗脂肪含量越低,平均剪切应力越大,挤压凝胶第一要素模型的胡克体弹性模量E1越小,其硬度越高,弹性和黏性越低;直链淀粉含量越高,支链淀粉含量越低,拉伸强度越高,松弛时间 τ1 越高,松弛时间 τ2 越低;松弛时间 τ 是方便米线品质控制中不可忽视的重要因素;建立的方便米线品质因素与原料大米成分间的交互效应方程具有较高的拟合度,此方程对于方便米线加工原料的选择具有指导意义。