Combination of high-speed countercurrent chromatography and reversed phase C18 chromatography for large-scale isolation of cyanidin-3-O-β-d-glucoside from black rice bran extract

Cyanidin-3-O-β-d-glucoside (C3G), the best known and most investigated anthocyanin, has many pharmacological properties. Black rice bran is an outstanding source for the separation of C3G due to its high C3G content and a simple anthocyanin composition. The present study described a large-scale isolation of C3G from the crude black rice bran extract by combining high-speed countercurrent chromatography (HSCCC) and reversed phase C₁₈ column chromatography. After HSCCC enrichment with the solvent system water-n-butanol-tert-butyl-methyl-ether (TBME)-acetonitrile-trifluoroacetic acid (TFA) (5:2:2:1:0.1%), the C3G content was concentrated about 16-fold and reached to 563.23 mg/g with a recovery 96.89%. Further purification was carried out using reversed phase C₁₈ column chromatography and 1.43 g of pure C3G (purity 94.38%) was obtained from 3.0 g of anthocyanin-enriched extract. The method is time-saving, low risk of sample denaturation, high sample recovery, and so is suitable for large-scale preparation of C3G for further studies of bioactivities.     

Value-addition of agricultural wastes for augmented cellulase and xylanase production through solid-state tray fermentation employing mixed-culture of fungi

Solid-state fermentation (SSF) was performed to evaluate the potential of agricultural residues for the production of cellulase and hemicellulase using individual and mixed cultures of Aspergillus niger and Trichoderma reseei. The maximum filter paper (FP) cellulase activity of 13.57 IU/gram dry substrate (gds), 22.89 IU/gds and 24.17 IU/gds and β-glucosidase activities of 21.69 IU/gds, 13.58 IU/gds and 24.54 IU/gds were obtained with wheat bran medium at 96 h incubation period with A. niger, T. reseei and mixed-cultures of A. niger and T. reseei, respectively. Mixed-culture SSF using rice straw supplemented with wheat bran in the ratio 3:2 resulted in higher FP cellulase, β-glucosidase, endoglucanase (CMCase) and xylanase activities, compared to the activities obtained using mono-cultures. Similarly, higher FP cellulase, β-glucosidase, CMCase and xylanase activities of 35.8 IU/gds (96 h), 33.71 IU/gds (96 h), 131.34 IU/gds (120 h) and 3106.34 IU/gds (120 h) were achieved in the tray fermentation using rice straw with wheat bran in the ratio of 3:2. Results of present investigation showed that higher cellulase activity and an optimal combination of cellulase and β-glucosidase can be achieved through mixed-culture SSF in trays. The approach of utilizing negative cost agricultural wastes through tray fermentation for cellulase and hemicellulase production is expected to serve the objectives of: (a) management of wastes which would otherwise cause environmental pollution problems; (b) production of hydrolytic enzymes at low cost and; (c) simple technique requiring no sophisticated instruments with practical applications.     

Nitrogen applications modify seed and oil yields and fatty acid composition of winter mustard

Winter mustard (Brassica juncea L.) is not a common crop in the Southeastern United States. With increased interest in biodiesel production, there has been corresponding interest in mustard in this region. The objective of this study was to evaluate the effect of N fertilization (0, 50, 100, 150 kg N ha⁻¹) on productivity, oil content, and oil composition of winter mustard ‘Pacific Gold’ grown at three locations in Mississippi (Stoneville, and two locations at Verona, namely Verona silt loam (Verona-SL) and Verona clay (Verona-C)). Nitrogen did not affect oil content (percent oil). Seed and oil yields (kg ha⁻¹) increased with N application relative to the unfertilized control. At the Verona-C location, the concentration of oleic acid was higher in the 50 kg N ha⁻¹ treatment. At Stoneville, linolenic acid concentration was higher in the 150 kg N ha⁻¹ and lower in the 100 kg/N ha⁻¹ treatment, while it was not different in the other treatments. Overall, the yield of the fatty acids (FA) palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, arachidic, eicosanoic, behenic, erucic, lignoceric, and nervonic acid increased with higher N rates (100 or 150 kg N/h). The highest yield of FA in the two Verona locations were achieved in the 100 kg N ha⁻¹, while greatest yield of FA at Stoneville was achieved in the highest N rate (150 kg N ha⁻¹). Means of mustard oil yields in our study in the higher fertility treatment ranged from 737 to 1094 kg ha⁻¹. This study demonstrated winter mustard production in Mississippi and possibly other areas in the Southeastern United States can be successful and could provide seed and oil yields comparable to yields from other production areas.     

Oil productivity and composition of sunflower as a function of hybrid and planting date

Sunflower (Helianthus annuus L.) is a potential cash crop for the southeastern United States for production of cooking oil or biodiesel. Two years (2006 and 2007) of experiments were conducted at each of five locations in Mississippi to evaluate the effect of planting date (April 20, May 20, and June 20), and hybrid (DKF3875, DKF2990, DKF3510, DKF3901, PR63M80, PR62A91, PR63A21, PR63M91, and PR64H41) on seed yield, oil content, and oil composition of sunflower. Seed oil concentration varied from 25 to 47%. The oleic acid concentration in the oil was greater than 85% for DKF3510 and PR64H41, above 65% for PR63M80 and PR63M91, and intermediate for the other hybrids. Total saturated fatty acids (TSFA) concentration in the oil (the sum of palmitic, stearic, arachidic, behenic, and lignoceric acids) ranged from 6.3 to 13.0%, with DKF3510, PR63M91, and PR64H41 having lower concentration of TSFA than the other hybrids. Mean seed yields ranged from 997 to 2096 kg ha⁻¹ depending on location. Mean oil yields at the five locations ranged from 380 to 687 kg ha⁻¹, and calculated biodiesel production ranged from 304 to 550 kg ha⁻¹. Seed and oil yields in this study suggest sunflower in Mississippi should be planted by the last week of May. Later planting (20 June) may significantly decrease both seed and oil yields in the non-irrigated system in Mississippi and in other areas of the southeastern United States with similar environmental conditions.     

Synthesis of Jatropha curcas oil-based biodiesel in a pulsed loop reactor

Jatropha curcas oil (JCO) has a high content of free fatty acids and has been used extensively as a feedstock in biodiesel production. In the present study, the transesterification reaction of JCO to Jatropha curcas methyl ester (biodiesel) was performed in a continuous pulsed loop reactor under atmospheric conditions. The JCO was pre-treated prior to the reaction to reduce the free fatty acid content to below 1% (w/w). The operating parameters of the loop reactor were optimised based on the conversion of the JCO to Jatropha curcas biodiesel and included reaction temperature, molar ratio of oil to MeOH, reaction time and oscillation frequency. The findings show that the highest reaction conversion of 99.7% (w/w) was achieved using KOH catalyst and 98.8% conversion was obtained using NaOCH₃ catalyst. The optimal operating conditions were a molar ratio of 6:1, an oscillation frequency of 6 Hz, temperature of 60 °C, feedstock FFA content of 0.5% (w/w) and only 10 min of reaction time. As a commercial commodity, the physical properties of biodiesel were analysed, and they compared well with the characteristics of fossil-based diesel fuel.     

Methyl ester of Sal oil (Shorea robusta) as a substitute to diesel fuel—A study on its preparation, performance and emissions in direct injection diesel engine

Many research reported vegetable oil as a potential substitute for diesel engines with its ester form known as biodiesel. The biodiesel can be prepared by different process using vegetable oil and alcohol. The common process used for biodiesel preparation is known as transesterification. This paper presents the transesterification of Sal oil (Shorea robusta) into Sal oil methyl ester (SOME) and its performance in direct injection diesel engine. Several process parameters such as catalyst quantity, molar ratio of alcohol, reaction temperature and reaction time were studied and the optimized process conditions are amount of catalyst (NaOH) - 0.25 wt%, alcohol (methanol) - 150% excess, reaction temperature - 65 °C and reaction time - 1.5 h. The studies with SOME as fuel in the direct injection diesel engine shows that the exhaust emissions such as CO, HC and NO_x are reduced by 25%, 45% and 12%, respectively compared to diesel without significant difference in thermal efficiency. Based on this study it is concluded that the SOME can be used as fuel without any modifications in the engine and hence this biodiesel can be a potential substitute to standard diesel fuel.     

Synthesis and characterization of hazelnut oil-based biodiesel

The demand for diesel fuel far exceeds the current and future biodiesel production capabilities of the vegetable oil and animal fat industries. New oilseed crops that do not compete with traditional food crop are needed to meet existing energy demands. Hybrid hazelnut oil is just such an attractive raw material for production of biodiesel. Hazelnut oil was extracted from hybrid hazelnuts and the crude oil was refined. Hazelnut oil-based biodiesel was prepared via the transesterification of the refined hazelnut oil with excess methanol using an alkaline catalyst. The effects of reaction temperature, time and catalyst concentration on the yield of diesel were examined, and selected physical and chemical properties of the biodiesel were evaluated. The biodiesel yield increased with increasing temperature from 25 to 65 °C and with increasing catalyst concentration from 0.1 to 0.7 wt%. The increase in yield with reaction time was nonlinear and characterized by an initial faster rate, followed by a slow rate. Hazelnut oil-based biodiesel had an average viscosity of 8.82 cP at 25 °C, which was slightly higher than that of the commercial soy-based diesel (7.92 cP at 25 °C). An approximate 12 °C higher onset oxidative temperature and a 10 °C lower cloud point of hazelnut oil biodiesel than those of its commercial soy counterpart indicated a better oxidative stability and flowability at low temperature. The average heat of combustion of hazelnut oil biodiesel was 40.23 kJ/g, and accounted for approximately 88% of energy content of diesel fuel. The fatty acid composition of hazelnut oil-based biodiesel was the same as the nature oil.     

茶籽油制备生物柴油工艺研究

以茶树[Camellia sinensis (L.) O. Kuntze]茶籽油为原料,研究了茶籽油甲酯化制备生物柴油的工艺条件。在单因素试验的基础上,选取反应温度、催化剂用量（占精炼油质量百分比）、反应时间和醇油摩尔比为影响因子,以酯交换率为响应值,应用Box-Behnken中心组合试验设计建立数学模型,进行响应面分析。结果表明,茶籽油制备生物柴油的最佳工艺条件为：反应温度58℃、催化剂用量1.05%、反应时间66min、醇油摩尔比9.7∶1。在此条件下,酯交换率达到98.73%。对生物柴油进行红外光谱和GC-MS分析,产品质量达到国家生物柴油标准。     

Formation,characterization, and glucose response in mice to rice starch with low digestibility produced by citric acid treatment

The optimum conditions for producing rice starch enriched in slowly digestible and resistant fractions by citric acid treatment determined by a response surface methodology (RSM) model equation, were: reaction temperature, 128.4 °C; reaction time, 13.8 h; and citric acid, 2.62 mmol/20 g starch. The slowly digestible and resistant starch fractions of the optimally acid-treated rice starch totalled 54.1%, which was 28.1% higher than the control. The slowly digestible and resistant fractions of the acid-treated rice starch did not differ significantly after heat treatment, whereas those of raw rice starch decreased by 49.6–63.8%, depending on the type of heat treatment (cooking at 100 °C or autoclaving). The slowly digestible fraction of the acid-treated starch increased by 8.9–14.2%. After autoclaving, the glucose response of the acid-treated starch was lower than untreated starch, but similar to that of Novelose 330. After heat treatment, the rate of blood glucose decrease was slower for the acid-treated starch than for Novelose 330. Compared to raw rice starch, the acid-treated starch exhibited increases in apparent amylose content, blue value, dextrose equivalent, cold-water solubility and transmittance, and decreases in wavelength of maximum absorbance, viscosity, and gel-forming ability.     

不同调酸剂对水稻育秧基质调酸效果及对秧苗生长的影响

育秧基质合适酸碱度(pH值)不仅有利于水稻正常生长,同时有利于预防水稻病害.因此,研究不同类型调酸剂对育秧基质的调酸效果,以及育秧过程中对水稻生长的影响很有必要.以我们自己研制的发酵基质作为研究对象,分别添加不同浓度的柠檬酸、稀硫酸、硫磺、磷酸二氢铵,测定基质的pH值和电导率(EC值).播种后12 d取样,结果发现:(...     

Jatropha curcas seed cake as substrate for production of xylanase and cellulase by Aspergillus niger FGSCA733 in solid-state fermentation

Jatropha curcas seed-cake was evaluated for use as a solid state fermentation substrate for production of cellulolytic and xylanolytic enzymes by Aspergillus niger. Supplementation of the seedcake with 10% thatch grass (Hyperrhaenia sp.) resulted in a fivefold increase in xylanase production. Ammonium chloride supplementation increased production of xylanase by 13%. Under the same conditions, cellulase production was not influenced by supplementation with grass or the nitrogen sources used. Maximum xylanase was produced at 25 °C whilst cellulase was maximally produced at 40 °C. Highest xylanase activity was obtained when the cultures had an initial pH of 3 whereas cellulase was maximally produced at an initial pH of 5. Under optimised conditions, 6087 U and 3974 U of xylanase and cellulase respectively were obtained per gram of substrate. Zymograms of crude enzyme extracts showed six active bands ranging from 20 kDa to 43 kDa for cellulase and a 31 kDa active band for xylanase.     

Dephytinization of wheat bran by fermentation with bakers' yeast,incubation with barley malt flour and autoclaving at different pH levels

Wheat bran is an important source of dietary fiber but also contains considerable amounts of phytic acid, which is known to impair mineral absorption. The present study was conducted to investigate the phytic acid reduction in coarse and fine wheat bran by fermentation with the different levels of bakers' yeast (3, 6 and 9%) for 8 h at 30 °C, incubation with the different levels of barley malt flour (2.5, 5.0, 7.5 and 10.0%) for 8 h at pH 5.2 and 55 °C, and autoclaving at the different pH levels (pH 5.0, 4.5, 4.0 and 3.5) adjusted with acetic acid for 2 h. The phytic acid content of the wheat bran was effectively reduced by all treatments, and the phytic acid lost was in the range of 88.4–96.9%. Without addition of yeast or malt flour, or autoclaving without pH adjustment, the phytic acid content of the bran samples was reduced at most to 44.9% of the initial amounts under the investigated conditions. Increasing the concentration of yeast or malt flour or decreasing the pH towards 3.5 did not enhance the phytic acid reduction. The most reduction occurred after 2 h of yeast fermentation and malt flour incubation, and after 30 min of autoclaving, which made up 92–98% of the total phytic acid loss. Extending the treatment periods contributed nominally to further increase in the phytic acid reduction, and the rate of the phytic acid loss decreased progressively.     

Process optimization for ethanol production from photoperiod-sensitive sorghum: Focus on cellulose conversion

Photoperiod-sensitive sorghum, as a competitive biomass for ethanol production, was investigated to develop an integrated process for improving ethanol yield. Response surface methodology was employed to study the relationship between pretreatment variables (including temperature, sulfuric acid concentration, and reaction time) and cellulose recovery, as well as efficiency of enzymatic hydrolysis (EEH) in the solid part. Recovery yield decreased and EEH increased as the pretreatment temperature, acidic concentration, and reaction time increased. A model was successfully developed to predict total glucose yield with a maximum value of 82.2%. Conditions of co-fermentation were also optimized, and the optimal ethanol yield was obtained with constant-temperature simultaneous saccharification and fermentation at 38 °C. Acetate buffer at a concentration of 50 mM was found helpful for increasing efficiency of enzymatic hydrolysis, as well as ethanol yield. The maximum ethanol yield was 0.21 g ethanol per dry mass at the conditions of 38 °C, 0.05 g yeast/L, and 50 mM acetate buffer. A complete cellulose balance was provided for the whole process.     

Regulation of tissue differentiation by plant growth regulators on tTCLs of Panax ginseng adventitious roots

Differentiated tissue in Panax ginseng cultures was found to be very efficacious for saponin production. In order to increase the yield of saponins and preserve culture stability we were testing different plant growth regulators (PGR) and auxin/cytokinin combinations to regulate a level of tissue differentiation. For this purpose we used transverse thin cell layers (tTCLs) of adventitious roots of Panax ginseng. Adventitious roots were cultivated in Shenk and Hildebrand (SH) liquid medium supplemented with IBA (24.6 μM). Callus formation and root multiplication of adventitious root tTCLs was evaluated after 4 and following 12 weeks of cultivation, respectively, on SH basal medium containing various auxins (3 mg l⁻¹) or cytokinins (0.2 or 0.02 mg l⁻¹) or their combinations. We found that kinetin (Kin) in combination with auxin benzo[b]selenienyl acetic acid (BSAA), naphthalene acetic acid or indole-3-butric acidis the best for biomass production and following root multiplication. These combinations were tested in previously selected most suitable large-scale system—a temporary immersion system RITA. The best saponin production (15.94 ± 1.89 mg g⁻¹ dry weight) and growth value (5.62 ± 0.34) was reached on medium containing BSAA and Kin combination.     

Synthesis of a base-stock for electrical insulating fluid based on palm kernel oil

This report presents a method for synthesizing base-stock for green industrial product from a vegetable oil with a high composition of unsaturated fatty acids. Epoxy methyl ester of palm kernel oil was synthesized from laboratory purified palm kernel oil using a two-step reaction and the products were used as a base-stock for green electrical insulation fluid. Epoxidized palm kernel oil was first prepared through epoxidation reaction involving purified palm kernel oil, acetic acid and hydrogen peroxide in the presence of amberlite as catalyst which lasted for 4 h. It was then followed by transesterification reaction involving the epoxidized product and methanol in the presence of sodium hydroxide as catalyst to synthesize the corresponding epoxy methyl ester. The thermal and electrical breakdown properties of the epoxy methyl ester demonstrated significantly improved properties for its use as raw material for bio-based industrial products such as electrical insulation fluids.     

Stabilization of Tarom and Domesiah cultivars rice bran: Physicochemical,functional and nutritional properties

Extrusion is a multi-step thermal process which has been extensively utilized in food preparations. Effects of temperature, moisture content and speed screw on the stabilization of rice bran were optimized using response surface methodology. Furthermore, the effect of extrusion processing on the physicochemical, nutritional and functional properties of Tarom and Domesiah cultivars stabilized rice bran (SRB) were studied. The colour of rice bran was improved by extrusion processing, but the protein content was reduced, which can be related to the denaturation of protein. Extrusion had also a reduction significant effect on the phyitic acid and vitamin E in stabilized rice bran. However, the content of vitamins B₂, B₃, B₅ and folic acid were remained unchanged, but the dietary fibre was enhanced which has beneficial health effect. In comparison with raw rice bran, water holding capacity was enhanced, but the oil absorption capacity was reduced. Foaming capacity and foaming stability of SRB was more than that of untreated rice bran. Although, they were less than that of rice bran protein concentrate/isolate. As a result, extrusion process improves some functional and nutritional properties of rice bran which are valuable for industrial applications and have potential as food ingredient to improve consumer health.     

Enzymatic hydrolysis of arabinoxylans from spelt bran and hull

Spelt bran and hull were submitted to enzymatic treatment during 1 h, 4 h and 24 h to release arabinoxylans (AXs). Several enzymatic commercial preparations with mainly endo-xylanase activity from Trichoderma reesei (Rohalase WL), Thermomyces lanuginosus (Pentopan Mono conc. BG), Bacillus subtilis (Belfeed B MP and Grindamyl Powerbake 900), Humicola insolens (Ultraflo L), Aspergillus aculeatus (Shearzyme 2×), A. aculeatus plus T. reesei (Shearzyme Plus) were tested and combined with a commercial cellulolytic preparation from T. reesei (Celluclast 1.5L). The xylanolytic activity of the commercial enzyme preparations was heterogeneous. The production of monomers by the different enzymes was evaluated on oat spelt xylan. Spelt bran and hull hydrolysates were analysed in regard to their content of total and free sugars. Monomers of glucose were mainly released with Celluclast, Ultraflo L and Shearzyme 2×. The percentage of released AXs ranged from 18.1% to 69.7% for bran and 0.5% to 6.4% for hull (% of the AXs originally present in the substrate). The solution of AXs hydrolysed by endoxylanases from spelt bran contained polysaccharides with a degree of polymerisation (DP) of 2–71 while those hydrolysed with endoxylanases plus Celluclast had DPs were 1–1164. The hydrolysates from spelt hull had much lower DPs (between 2 and 17).     

Soil properties affected by combinations of soil solarization and organic amendment

A field experiment was conducted to investigate the effects on soil properties of solarization combined with rice bran additive. The treatments included control (bare soil), black polyethylene film mulch (BM), clear polyethylene film mulch (CM), rice bran mixture (RBMx), rice bran mulch (RBM), rice bran mixture and black polyethylene film mulch (RBMx + BM), rice bran and black polyethylene film mulch (RBM + BM), rice bran mixture and clear polyethylene film mulch (RBMx + CM), and rice bran and clear polyethylene film mulch (RBM + CM). Initially, 15 kg m⁻² of rice bran was applied to each treatment as the organic amendment. A cover of black or clear polyethylene film was then used to begin the soil solarization process. Approximately 49 days later, the polyethylene film was removed and five broccoli seedlings per replication were planted 6 weeks later. The combination of rice bran mixture with the clear/transparent polyethylene film mulch (RBMx + CM) raised both the mean and maximum soil temperatures over 0–30 cm of soil depth. Maximum soil temperature under the RBMx + CM treatment was the highest of all treatments (77°C) and were about 32°C greater than under the control, while it was only 52°C under RBM + BM treatment. Combining rice bran with clear/transparent polyethylene film mulch during the soil solarization process is recommended for raising soil temperatures. Further investigation over a longer period of soil solarization process with rice bran–organic amendment and polyethylene film mulch combinations is required to more accurately determine their effects on soil properties and crop production.     

Preparation of biodiesel from Idesia polycarpa var. vestita fruit oil

The feasibility of producing biodiesel from Idesia polycarpa var. vestita fruit oil was studied. A methyl ester biodiesel was prepared from refined I. polycarpa fruit oil using methanol and potassium hydroxide (KOH) in an alkali-catalyzed transesterification process. The experimental variables investigated in this study were catalyst concentration (0.5–2.0 wt.% of oil), methanol/oil molar ratio (4.5:1 to 6.5:1), temperature (20–60 °C) and reaction time (20–60 min). A maximum yield of over 99% of methyl esters in I. polycarpa fruit oil biodiesel was achieved using a 6:1 molar ratio of methanol to oil, 1.0% KOH (% oil) and reaction time for 40 min at 30 °C. The properties of I. polycarpa fruit oil methyl esters produced under optimum conditions were also analyzed for specifications for biodiesel as fuel in diesel engines according to China Biofuel Systems Standards. The fuel properties of the I. polycarpa fruit oil biodiesel obtained are similar to the No. 0 light diesel fuel and most of the parameters comply with the limits established by specifications for biodiesel.     

Effects of visible light irradiation on the oxidative stability in rice bran

Raw rice bran was treated with or without visible light exposure at room temperature or stored at 40 °C in the dark for 10 days and rice bran oil (RBO) was recovered from each rice bran. Headspace oxygen content from rice bran and conjugated dienoic acid (CDA) value, acid value, content of γ-oryzanol, and fluorescence intensity in RBO were analyzed to determine the effects of visible light on the oxidative stability in rice bran. Headspace oxygen content in visible light irradiated rice bran (RBL) decreased by 12.8% for 10 days while those in the dark (RBD) and stored at 40 °C (RBT) decreased by 5.87 and 5.35%, respectively, implying visible light irradiation accelerates the consumption of oxygen. CDA values in RBO from RBL were significantly higher than those in RBO from RBD and RBT (p < 0.05). However, acid values in RBO were not significantly different among samples (p > 0.05). Both γ-oryzanol content and fluorescence intensity in RBO from RBL were significantly lower than those in RBO from RBD and RBT (p < 0.05). Fluorescence intensity, which is related to the content of chlorophylls, decreased in samples under light only, implying that chlorophyll photosensitization may play important roles in the acceleration of lipid oxidation in rice bran.