Preparation of a rice bran enzymatic extract with potential use as functional food

The production, stabilization, by enzymatic treatment, physicochemical composition, and biological properties (including the anti-proliferative activity), of a water-soluble rice bran enzymatic extract (RBEE) are described. The main component of RBEE is proteins (38.1%) – in the form of peptide and free amino acids – having a 6% content of sulfur amino acids. The second component is fat (30.0%), with oleic and linoleic acids as the major components, and 1.2 mg/g of γ-oryzanol. Carbohydrates (14.2%) are comprised mainly of slowly absorbed carbohydrates. Preliminary studies on the anti-proliferative effect of RBEE on leukemia tumor cell growth in vitro are also reported. This property makes RBEE potentially useful as a functional food for the treatment and prevention of chronic pathological states associated with abnormal proliferation of cells, as is the case with cancer.     

Partial extraction method for the rapid analysis of total lipids and γ-oryzanol contents in rice bran

Total lipids and γ-oryzanol in rice bran were determined by a partial extraction method. The results agreed well with the conventional total extraction methods. The proposed method uses fewer hazardous organic solvents, takes a shorter extraction time and requires no special extraction apparatus. Total lipids and γ-oryzanol in nine rice bran varieties were analysed by the developed technique. Daw Dum 5647 had the highest total lipids and γ-oryzanol while the lowest content was found in KD XBT 313-19-1-1 and SP XBT 43-7, respectively. The adsorption coefficient (K_d) of the lipids and γ-oryzanol, between hexane and bran, at 30 °C are between 1.16 and 2.00 and 2.02 and 2.65, respectively (depending on the moisture content of the bran). From the K_d values, it was estimated that about 92–95% of the lipids and 95–96% of the γ-oryzanol were extracted into hexane at a 10:1 (v/w) ratio of hexane to bran. The effect of solvents on the extraction of γ-oryzanol from rice bran was also studied. It was found that isopropanol was the most suitable solvent for extraction and determination of γ-oryzanol in rice bran. It showed better agreement with the total extraction method.     

Quality Changes and Tocopherols and γ-Orizanol Concentrations in Rice Bran Oil During the Refining Process

The quality changes and the concentrations of tocopherols and γ-oryzanol, during successive steps of rice bran oil refining (RBO), were studied. For this purpose, samples of crude, degummed, neutralized, bleached, dewaxed and deodorized RBO were taken from an industrial plant and analyzed. The moisture, pH, acidity, peroxide value and unsaponifiable matter, were determined. The fatty acid composition was evaluated by GC, and the concentrations of tocopherols and γ-oryzanol were determined using HPLC with fluorescence and UV–Vis detection, respectively. To identify γ-oryzanol components, fractions of the HPLC eluant were collected and analyzed using mass spectrometry. Oil refining reduced the peroxide value and acidity to 1 and 3% of the values obtained in crude RBO, respectively. The fatty acid composition were not significantly altered during refining. The concentrations of the tocopherols in RBO followed the order α > (β + γ) > δ. The total concentration of tocopherols was 26 mg/100 g, and remained practically unaltered during refining. Up to nine components were distinguished in γ-oryzanol. After collecting the elution fractions, up to six components were identified by electrospray mass spectrometry. Refining reduced the total concentration of γ-oryzanol to 2% of its initial value.     

Recovery of γ-oryzanol from biodiesel residue

γ-Oryzanol has important applications in food, cosmetic and pharmaceutical industries. The objective of this investigation is to isolate γ-oryzanol from residue obtained during the production of biodiesel from rice bran oil. Using rice bran oil as the feedstock, the content of γ-oryzanol could be raised to 16% by a series of steps, which include degumming and dewaxing, acid-catalyzed esterification and vacuum distillation. More than 95% low-boiling point components, such as free fatty acid and fatty acid methyl ester (biodiesel), were obtained as the distillate. After applying solvent extraction to the residue, γ-oryzanol content was increased from 16 to 35% with a recovery of 88%. Subsequent use of soxhlet extraction raised γ-oryzanol content to 47% with a recovery of 97%. Finally, after applying silica gel column chromatography, γ-oryzanol content was 83.79% with a recovery of 81.75%. The overall recovery was 69.82%.     

Enzymatic hydrolysis of steryl ferulates and steryl glycosides

Steryl ferulates (SF) and steryl glycosides (SG) are phytosterol conjugates found characteristically in cereals. Currently, little is known about their properties with respect to enzymatic hydrolysis. SF and SG were extracted and purified from rye and wheat bran. Their percentages of hydrolysis with different enzymes were studied using normal phase HPLC with UV detection for steryl ferulates and evaporative light scattering detection for steryl glycosides. Steryl ferulates were hydrolysed by mammalian digestive steryl esterases. It was further demonstrated that a mixture of steryl ferulates from rye and wheat was hydrolysed much more effectively than a steryl ferulate mixture from rice (commonly known as γ-oryzanol), suggesting greater bioavailability in non-rice steryl ferulates. Steryl glycosides were hydrolysed by a commercial microbial β-glucosidase preparation (cellobiase), but were not effectively hydrolysed by two other highly purified β-glucosidases. These results demonstrate for the first time the potential use of enzymes as a replacement for acid hydrolysis in analytical procedures for SG and also provide insights about the potential bioavailability of these sterol derivatives in human digestive systems. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Composition,industrial processing and applications of rice bran γ-oryzanol

Rice bran oil (RBO) (20–25 wt% in rice bran) is a unique rich source of commercially-important bioactive phytochemicals, most of them of interest in nutrition, pharmacy and cosmetics. The unsaponifiable constituents of RBO include mainly tocols (vitamin E, 0.10–0.14%) and γ-oryzanol (esters of trans-ferulic acid with sterols and triterpenic alcohols, 0.9–2.9%). The following topics concerning γ-oryzanol are reviewed: analytical methods for characterisation and determination; influence of genetic and environmental factors on the composition of rice bran; extraction approaches, including supercritical CO₂ and subcritical water; and biomedical and industrial applications, including food and pharmaceuticals. Concentration ranges of γ-oryzanol, tocopherols and tocotrienols found in rice bran and RBO from different varieties and geographical areas are summarised. This review focuses on the 2003–2008 period, where an average of 13–14 references per year were published; however, some relevant work reported during the 1998–2002 period is also briefly commented upon.     

Effects of the extraction conditions on the yield and composition of rice bran oil extracted with ethanol—A response surface approach

Rice bran oil was obtained from rice bran by solvent extraction using ethanol. The influence of process variables, solvent hydration (0-24% of water, on mass basis), temperature (60-90 °C), solvent-to-rice bran mass ratio (2.5:1 to 4.5:1) and stirrer speed (100-250 rpm) were analysed using the response surface methodology.The extraction yield was highly affected by the solvent water content, and it varied from 8.56 to 20.05 g of oil/100 g of fresh rice bran (or 42.7-99.9% of the total oil available) depending on the experimental conditions. It was observed that oryzanol and tocols behave in different ways during the extraction process. A larger amount of tocols is extracted from the solid matrix in relation to γ-oryzanol. It was possible to obtain values from 123 to 271 mg of tocols/kg of fresh rice bran and 1527 to 4164 mg of oryzanol/kg of fresh rice bran, indicating that it is feasible to obtain enriched oil when this renewable solvent is used. No differences in the chemical composition of the extracted oils were observed when compared to the data cited in the literature.     

γ-Oryzanol and tocopherol contents in residues of rice bran oil refining

Rice bran oil (RBO) contains significant amounts of the natural antioxidants γ-oryzanol and tocopherols, which are lost to a large degree during oil refining. This results in a number of industrial residues with high contents of these phytochemicals. With the aim of supporting the development of profitable industrial procedures for γ-oryzanol and tocopherol recovery, the contents of these phytochemicals in all the residues produced during RBO refining were evaluated. The samples included residues from the degumming, soap precipitation, bleaching earth filtering, dewaxing and deodorisation distillation steps. The highest phytochemical concentrations were found in the precipitated soap for γ-oryzanol (14.2 mg g⁻¹, representing 95.3% of total γ-oryzanol in crude RBO), and in the deodorisation distillate for tocopherols (576 mg 100 g⁻¹, representing 6.7% of total tocopherols in crude RBO). Therefore, among the residues of RBO processing, the deodorisation distillate was the best source of tocopherols. As the soap is further processed for the recovery of fatty acids, samples taken from every step of this secondary process, including hydrosoluble fraction, hydrolysed soap, distillation residue and purified fatty acid fraction, were also analyzed. The distillation residue left after fatty acid recovery from soap was found to be the best source of γ-oryzanol (43.1 mg g⁻¹, representing 11.5% of total γ-oryzanol in crude RBO).     

Nanofiltration process for the nutritional enrichment and refining of rice bran oil

Crude rice bran is a natural source of γ-oryzanol, a nutritionally valuable phytochemical with antioxidant properties. In the present paper the refining and γ-oryzanol enrichment of rice bran oil was investigated through solvent extraction optimization and nanofiltration processing. Several solvent resistant nanofiltration membranes were screened and successfully applied in a two step membrane cascade with fluxes between 39 and 53 L m⁻² h⁻¹. A first membrane stage operation provided the separation between glycerides and γ-oryzanol, promoting the oil enrichment in this phytochemical. In the second membrane stage the oil could be refined to acceptable consumption levels (FFA < 0.20 wt.%) and its γ-oryzanol content was further enhanced. Overall, the integrated process provided a RBO γ-oryzanol enrichment from 0.95 to 4.1 wt.% in oil, which corresponded to more than a two fold increase in the oil’s antioxidant capacity. These results demonstrate the potential of organic solvent nanofiltration as a technology to enrich and refine oil based products.     

Lipid Components of North American Wild Rice (<Emphasis Type="Italic">Zizania palustris</Emphasis>)

The content and composition of fatty acids, sterols, tocopherols, and γ-oryzanol in wild rice (Zizania palustris) grown in North America were compared with those in regular brown rice (Oryza sativa L.). The lipid content of wild rice ranged from 0.7 to 1.1%, compared with 2.7% in regular brown rice. The lipids of wild rice comprised mainly linoleic (35–37%) and linolenic (20–31%) acids. Other fatty acids included palmitic (14.1–18.4%), stearic (1.1–1.3%), and oleic (12.8–16.2%). Wild rice lipids contained very large amounts of sterols, ranging from 70 g/kg for a Saskatchewan sample to 145 g/kg for Minnesota Naturally Grown Lake and River Rice. The main sterols found in an unsaponified fraction were: campesterol (14–52%), β-sitosterol (19–33%), Δ⁵-avenasterol (5–12%), and cycloartenol (5–12%). Some of sterols, γ-oryzanols, were present as the phenolic acid esters; the amount ranged from 459 to 730 mg/kg in wild rice lipids. The largest amounts of tocopherols and tocotrienols, 3682 and 9378 mg/kg, were observed in North Western Ontario wild rice samples, whereas the lowest were 251 mg/kg in an Athabasca Alberta sample and 224 mg/kg in regular long-grain brown rice. The α isomer was the most abundant among tocopherols and tocotrienols. The results of this study showed that wild rice lipids contain large amounts of nutraceuticals with proven positive health effects.