Optimizing Extraction of Zein and Glutelin‐Rich Fraction During Sequential Extraction Processing of Corn

This study was conducted to improve yields and qualities of corn protein co‐products produced by the sequential extraction process (SEP), a process using ethanol to fractionate corn in producing fuel ethanol. A two‐stage extraction protocol was evaluated to recover zein and subsequently recover a glutelin‐rich fraction (GRF). After the simultaneous oil‐extraction and ethanol‐drying step of SEP, zein was extracted from the anhydrous‐ethanol‐defatted, flaked corn by using 70% (v/v) ethanol at 60°C for 1.5 hr in a shaking water bath. Zein was recovered by ultrafiltering and then drying in a vacuum‐oven. Zein yield was 65% of the available zein in the flaked corn. SDS‐PAGE band patterns of the recovered zein closely resembled that of commercial zein. After zein extraction, the GRF was extracted using 45% ethanol and 55% 0.1M NaOH at 55°C for 2 hr. The extract was concentrated by ultrafiltration and then freeze‐dried. GRF yield was ≈65% of the available protein. Freeze‐dried GRF contained 90% crude protein (db), which classified the protein as a protein isolate. As with the protein concentrate from the original SEP, the GRF isolate was highly soluble in water at pH ≥ 7, had good emulsifying and foaming properties, formed stable emulsions, and was heat‐stable.     

Sequential Extraction Processing of High‐Oil Corn

The Sequential Extraction Process (SEP), a process that uses ethanol to fractionate corn into high‐value co‐products when producing fuel ethanol, was evaluated using high‐oil corn (HOC). Oil and protein recoveries, ethanol‐drying capability, and oil and protein properties were compared with those produced from normal soft dent corn (SDC) using SEP. Moisture adsorption capacities (≈24 g of water/kg of corn) and oil recoveries (>95%) were nearly identical for both corn types. However, oil yield from HOC (7.1 g of oil/100 g of corn) was 65% more than that of SDC (4.3 g of oil/100 g of corn). HOC crude oil was less red and contained lower free fatty acid and phosphatide contents than did SDC oil. HOC zein contained higher crude protein content (86% db) than did SDC zein (79% db). The freeze‐dried glutelin‐rich fractions (GRF) from both types of corn contained >90% (db) crude protein, which meet the criteria of protein isolates. SEP GRF protein isolates had >85% protein soluble in water at pH ≥ 7. Both GRF protein isolates from HOC and SDC were heat‐stable, had good emulsifying capacities, and produced highly stable emulsions. They also had substantial foaming capacities, but the foam from HOC GRF protein isolate was significantly more stable than the foam from SDC GRF protein isolate. SEP is a suitable process for recovering oil and protein products from HOC corn when producing fuel ethanol and, if competitively priced, HOC is a preferred feedstock to SDC because of higher yields and qualities of valuable co‐products when using HOC.     

Quality Comparison of Rice Bran Oil Extracted with d‐Limonene and Hexane

d‐Limonene, a safe agricultural by‐product, was used to extract rice bran oil and compared against hexane, a petroleum product widely used as a solvent for extracting edible oil. The yield of crude rice bran oils extracted with both solvents in percentage by weight was obtained. The quality of crude rice bran oil was analyzed. The yield and quality of crude rice bran oil from the limonene‐based solvent extraction were almost equivalent to those from the hexane‐based operation. The optimum solvent‐to‐rice bran ratio and extraction time required for d‐limonene extraction of oil, based primarily on crude rice bran oil yield, have been determined to be 5:1 and 0.5 hr, respectively. Despite the absence of antioxidants during the limonene recovery step with vacuum evaporation, the quantity of the oxidation products in the recovered limonene was <1% (wt) of the original limonene solvent. The application of d‐limonene solvent as an alternative to hexane in edible oil extraction could potentially eliminate the safety, environmental, and health issues associated with the use of hexane.     

Selected Factors Affecting Crude Oil Analysis of Distillers Dried Grains with Solubles (DDGS) as Compared with Milled Corn

With increasing production of distillers dried grains with solubles (DDGS), both fuel ethanol and animal feed industries are demanding standardized protocols for characterizing quality. AOCS Approved Procedure (Am 5‐04) was used for measuring crude oil content in milled corn and resulting DDGS. Selected factors, including sample type (milled corn, DDGS), sample origin (ethanol plant 1, 2, 3), sample particle size (original matrix, <0.71 mm, <0.50 mm mesh opening; the last two materials were obtained by grinding and sieving), solvent type (petroleum ether, hexane), extraction time (30, 60 min), and postextraction drying time (30, 60 min) were investigated by a complete factorial design. For milled corn, only sample origin and extraction time had significant effects (P < 0.05) on crude oil values measured, but for DDGS, besides those two factors, sample particle size, solvent type, and drying time also had significant effects. Among them, the particle size of DDGS had the most effect. On average, measured oil content in DDGS ranged from 11.11% (original matrix) to 12.12% (<0.71 mm) and to 12.55% (<0.50 mm). For measuring the crude oil content of DDGS, particle size reduction, 60 min of extraction, and 60 min of drying are recommended. Regardless of the underlining factors, the method was very repeatable (standard errors <0.05). The observed particle size effect on crude oil analysis of DDGS suggests the need for similar confirmations using other analytical methods.     

乙醇水溶液提取玉米胚芽油的工艺优化

为了解决水酶法提取玉米胚芽油生产成本高、提取时间长的缺点,该文采用乙醇水溶液作为提取剂提取玉米胚芽油。通过对粒径、料液比、温度、乙醇体积分数、p H值和时间等条件对油在油相、水相和渣相中分布的研究发现,物料粒径和乙醇体积分数对提高清油得率具有显著(P0.05)的影响,而提取时间对清油得率的影响最小(P0.05)。在单因素试验的基础上通过正交试验,得出乙醇水溶液提取玉米胚芽油的最佳工艺参数为:物料细粉4次(此时粒径为49.18μm)、料液比1∶7 g/m L、温度70℃、乙醇体积分数30%、p H值9.0、提取时间2 h。在该条件下,清油的得率为94.05%±0.32%,水相含油量为3.49%±0.77%,渣相含油量为2.55%±0.82%。分析乙醇水溶液提取的玉米胚芽毛油酸价、过氧化值和含水率等指标发现,该毛油的质量优于国标规定的玉米原油,并且和压榨一级成品油指标接近,只需要经过简单精炼就可以达到食用油要求。研究结果为乙醇水溶液工业化生产玉米胚芽油提供参考。     

Effect of Alternative Milling Techniques on the Yield and Composition of Corn Germ Oil and Corn Fiber Oil

The effects of alternative corn wet‐milling (intermittent milling and dynamic steeping (IMDS), gaseous SO₂ and alkali wet‐milling) and dry grind ethanol (quick germ and quick fiber with chemicals) production technologies were evaluated on the yield and phytosterol composition (ferulate phytosterol esters, free phytosterols, and fatty acyl phytosterol esters) of corn germ and fiber oil and compared with the conventional wet‐milling process. Small but statistically significant effects were observed on the yield and composition of corn germ and fiber oil with these alternative milling technologies. The results showed that the germ and fiber fractions from two of the alternative wet‐milling technologies (the gaseous SO₂ and the IMDS) had, for almost all of the individual phytosterol compounds, either comparable or signficantly higher yields compared with the conventional wet‐milling process. Also, both of the modified dry grind ethanol processes (the quick germ and quick fiber) with chemicals (SO₂ and lactic acid) can be used as a new source of corn germ and fiber and can produce oils with high yields of phytosterols. The alkali wet‐milling process showed significantly lower yields of phytosterols compounds in germ but showed significantly higher yield of free phytosterols, fatty acyl phytosterol esters and total phytosterols in the fiber fraction.     

Comparison of oil and phytosterol levels in germplasm accessions of corn, teosinte, and Job's tears

Seeds of 49 accessions of corn (Zea mays ssp. mays), 9 accessions of teosinte (Zea species that are thought to be ancestors and probable progenitors to corn), and 3 accessions of Job's tears (Coix lacryma), obtained from a germplasm repository, were ground and extracted with hexane. Whole kernel oil yields and levels of four phytonutrients (free phytosterols, fatty acyl phytosterol esters, ferulate phytosterol esters, and gamma-tocopherol) in the oils were measured. Among the seeds tested, oil yields ranged from 2.19 to 4.83 wt %, the levels of ferulate phytosterol esters in the oil ranged from 0.047 to 0.839 wt %, the levels of free phytosterols in the oil ranged from 0.54 to 1.28 wt %, the levels of phytosterol fatty acyl esters in the oil ranged from 0.76 to 3.09 wt %, the levels of total phytosterols in the oil ranged from 1.40 to 4.38 wt %, and the levels of gamma-tocopherol in the oil ranged from 0.023 to 0.127 wt %. In general, higher levels of all three phytosterol classes were observed in seed oils from accessions of Zea mays ssp. mays than in seed oils from accessions of the other taxonomic groups. The highest levels of gamma-tocopherol were observed in teosinte accessions.     

Neutral Lipids in Free,Bound, and Starch Lipid Extracts of Flours,Sourdough, and Portuguese Sourdough Bread Determined by NP‐HPLC‐ELSD

A detailed analysis was developed, focused on the neutral lipids (NL) in free (FL), bound (BL), and starch lipid (SL) extracts of maize and rye flours, sourdough, and broa (a traditional bread manufactured in Portugal). Selective sequential extraction of said lipids with hexane at 20°C, water‐saturated n‐butanol at 20°C, and n‐propanol‐water (3:1, v/v) at 100°C was performed to clean the lipid extracts from extraneous impurities, and isolation thereof from glyco‐ and phospholipids was by solid phase extraction of NL; these classes were then quantitatively assayed by HPLC, using evaporative light scattering detection, with calibration curves prepared with standard mixtures of NL. The BL and SL contents in the original flours increased and that of FL decreased throughout the fermentation and baking processes. The dominant NL class was not the same in all lipid extracts; the highest concentrations of triacylglycerols and the lowest concentrations of free fatty acids were detected in FL—with the former accounting for 82, 76, and 71% of the total FL in flours, sourdough, and bread, respectively. Triacylglycerols and free fatty acids also accounted for the highest concentrations found in BL: these, together with diacylglycerols, contributed up to 84% of the total neutral BL. High levels of free fatty acids and low levels of the remaining NL classes were typically found in SL: free fatty acids, triacylglycerols, sterol esters, and diacylglycerols accounted for ≈90% of the total SL.     

Selective enzyme-mediated extraction of capsaicinoids and caratenoids from chili guajillo puya (Capsicum annum L.) using ethanol as solvent

The selective extraction of capsaicinoids and carotenoids from chili guajillo "puya" flour was studied. When ethanol was used as solvent, 80% of capsaicinoids and 73% of carotenoids were extracted, representing an interesting alternative for the substitution of hexane in industrial processes. Additionally, when the flour was pretreated with enzymes that break the cell wall and then dried, extraction in ethanol increased to 11 and 7% for carotenoid and capsaicinoid, respectively. A selective two-stage extraction process after the treatment with enzymes is proposed. The first step uses 30% (v/v) ethanol and releases up to 60% of the initial capsaicinoids, and the second extraction step with industrial ethanol permits the recovery of 83% of carotenoids present in the flour.     

Comparison of Modified Dry‐Grind Corn Processes for Fermentation Characteristics and DDGS Composition

Three different modified dry‐grind corn processes, quick germ (QG), quick germ and quick fiber (QGQF), and enzymatic milling (E‐Mill) were compared with the conventional dry‐grind corn process for fermentation characteristics and distillers dried grains with solubles (DDGS) composition. Significant effects were observed on fermentation characteristics and DDGS composition with these modified dry‐grind processes. The QG, QGQF, and E‐Mill processes increased ethanol concentration by 8–27% relative to the conventional dry‐grind process. These process modifications reduced the fiber content of DDGS from 11 to 2% and increased the protein content of DDGS from 28 to 58%.     

Radical scavenging activity of black cumin (Nigella sativa L.), coriander (Coriandrum sativum L.), and niger (Guizotia abyssinica Cass.) crude seed oils and oil fractions

Crude vegetable oils are usually oxidatively more stable than the corresponding refined oils. Tocopherols, phospholipids (PL), phytosterols, and phenols are the most important natural antioxidants in crude oils. Processing of vegetable oils, moreover, could induce the formation of antioxidants. Black cumin (Nigella sativa L.), coriander (Coriandrum sativum L.), and niger (Guizotia abyssinica Cass.) crude seed oils were extracted with n-hexane and the oils were further fractionated into neutral lipids (NL), glycolipids (GL), and PL. Crude oils and their fractions were investigated for their radical scavenging activity (RSA) toward the stable galvinoxyl radical by electron spin resonance (ESR) spectrometry and toward 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical by spectrophotometric method. Coriander seed oil and its fractions exhibited the strongest RSA compared to black cumin and niger seed oils. The data correlated well with the total content of polyunsaturated fatty acids, unsaponifiables, and PL, as well as the initial peroxide values of crude oils. In overall ranking, RSA of oil fractions showed similar patterns wherein the PL exhibited greater activity to scavenge both free radicals followed by GL and NL, respectively. The positive relationship observed between the RSA of crude oils and their color intensity suggests the Maillard reaction products may have contributed to the RSA of seed oils and their polar fractions. The results demonstrate the importance of minor components in crude seed oils on their oxidative stability, which will reflect on their food value and shelf life. As part of the effort to assess the potential of these seed oils, the information is also of importance in processing and utilizing the crude oils and their byproducts.     

Phytochemical, antioxidant, and antiproliferative properties of seed oil and flour extracts of Maryland-grown tobacco cultivars

To determine the possible alternative use of tobacco, the seeds representing seven Maryland tobacco cultivars were investigated for their phytochemical, antioxidant, and antiproliferative properties. Tobacco seed oils were extracted by the Soxhlet method, and analyzed for their yield, density, refractive index, fatty acid profiles, and tocopherol profile. The defatted flours were extracted in 50% acetone and 80% ethanol. The tobacco seed oil and flour extracts were analyzed for total phenolic contents (TPC) and scavenging capacities against peroxyl, hydroxyl and 2,2-diphenyl-1-picryhydrazyl (DPPH) radicals. The fatty acid compositions of phospholipids and the protein content of the flours were also analyzed. In addition, oil and flour extracts of varieties MD609 and MD609LA were evaluated for their antiproliferative effects on HT-29 human colon cancer cells. All of the tested extracts significantly inhibited HT-29 cell proliferation except that from MD609 oil. The data from this study suggest the potential alternative use of tobacco seeds in developing natural antioxidants and antiproliferative agents for improving human health.     

Study of the main constituents of some authentic walnut oils

This paper describes the composition of walnut oils obtained from nuts collected from seven countries that are major suppliers of walnut oil. Oils were extracted from the nuts using small-scale industry pressing equipment and analyzed using standard methods for fatty acids, fatty acids in the triacylglycerol 2-position, tocopherols and tocotrienols, triacylglycerols, sterols, steradienes, and iodine value. Values for the composition of the sterols, triacylglycerols, fatty acids, iodine value, and tocopherol composition were generally in good agreement with the results of previous similar surveys. Tocotrienols were not detected in any sample. Steradienes (stigmastadiene, campestadiene, stigmastatriene, and campestatriene) were not detected in any oil.     

Determination of phospholipids in olive oil by 31P NMR spectroscopy

A nondestructive analytical method based on NMR spectroscopy was developed for the determination of phospholipids in olive oil. The phospholipids extracted from virgin olive oil with a mixture of ethanol/water (2:1 v/v) were identified and quantified by high resolution (31)P NMR spectroscopy. The main phospholipids found in olive oil were phosphatidic acid, lyso-phosphatidic acid, and phosphatidylinositol. Validation of the (31)P NMR methodology for quantitative analysis of phospholipids in olive oil was performed. Sensitivity was satisfactory with detection limits of 0.25-1.24 mumol /mL. In addition, the composition of fatty acids in phospholipids model compounds and those in olive oil samples was estimated by employing one- and two-dimensional (1)H NMR. The results indicated that the fatty acid composition in phospholipids and triacylglycerols of olive oil was similar.     

Development of a rapid method for the sequential extraction and subsequent quantification of fatty acids and sugars from avocado mesocarp tissue

Methods devised for oil extraction from avocado (Persea americana Mill.) mesocarp (e.g., Soxhlet) are usually lengthy and require operation at high temperature. Moreover, methods for extracting sugars from avocado tissue (e.g., 80% ethanol, v/v) do not allow for lipids to be easily measured from the same sample. This study describes a new simple method that enabled sequential extraction and subsequent quantification of both fatty acids and sugars from the same avocado mesocarp tissue sample. Freeze-dried mesocarp samples of avocado cv. Hass fruit of different ripening stages were extracted by homogenization with hexane and the oil extracts quantified for fatty acid composition by GC. The resulting filter residues were readily usable for sugar extraction with methanol (62.5%, v/v). For comparison, oil was also extracted using the standard Soxhlet technique and the resulting thimble residue extracted for sugars as before. An additional experiment was carried out whereby filter residues were also extracted using ethanol. Average oil yield using the Soxhlet technique was significantly (P < 0.05) higher than that obtained by homogenization with hexane, although the difference remained very slight, and fatty acid profiles of the oil extracts following both methods were very similar. Oil recovery improved with increasing ripeness of the fruit with minor differences observed in the fatty acid composition during postharvest ripening. After lipid removal, methanolic extraction was superior in recovering sucrose and perseitol as compared to 80% ethanol (v/v), whereas mannoheptulose recovery was not affected by solvent used. The method presented has the benefits of shorter extraction time, lower extraction temperature, and reduced amount of solvent and can be used for sequential extraction of fatty acids and sugars from the same sample.     

Importance of Protein‐Rich Components in Emulsifying Properties of Corn Fiber Gum

Purified corn fiber gum (CFG‐F) isolated from fine (kernel endosperm‐derived) corn fiber that contained ≈2% residual protein was extracted with 70% aqueous ethanol. The aqueous ethanol extract (AEE), which contained 19.5% of the total CFG, contained a high percentage of the proteinaceous material present in the original gum sample. The AEE gum contained 6.81% protein by weight. The residue (R), which constituted 66% of the total CFG‐F, contained only 0.55% of protein. The emulsifying properties of R and AEE in a model oil‐in‐water emulsification system were studied by measuring turbidity after 1, 2, and 3 weeks, particle size after 4 weeks, and by confocal laser scanning microscopy after three months of storage at room temperature. These gums were compared with the standard well‐known emulsifiers native acacia gum (NAG) and modified acacia gum (MAG). The results indicate that although AEE contains protein‐rich components, it is not as good an emulsifier as the residue which contains only 0.55% of protein. However, emulsions prepared with the whole (unfractionated) CFG‐F under similar conditions were more stable showing higher turbidity and smaller particles size than those prepared with either R or AEE.     

亚麻籽双液相多级逆流萃取工艺模拟试验

为了考察双液相技术萃取亚麻籽的工业化效果,应用模拟试验探讨了多级逆流萃取工艺应用于正己烷-乙醇-水三元双液相亚麻籽萃油脱氰苷生产的可行性。结果表明,在料烷比1︰5 (m/V),料醇比1︰2 (m/V),温度55℃,时间 30 min,乙醇质量分数85% (m/m),NaOH添加量为乙醇相0.05% (m/V)的条件下,实验室串级模拟四级逆流萃取达到工艺要求,亚麻粕中残油量小于1%,氰苷残余小于0.7 mg/kg,多级逆流接触法萃取流程适宜于亚麻籽双液相萃油脱氰苷生产工艺。     

Enzymatic Process for Corn Dry‐Grind High‐Solids Fermentation

A modified dry‐grind process that combined the use of conventional amylases (glucoamylase [GA]), phytase, and granular starch hydrolyzing enzymes (GSHE) to achieve low liquefaction viscosities and low glucose concentrations during simultaneous saccharification and fermentation (SSF) with a high slurry solids content (>33% w/w) was developed. Doses of GSHE and GA were optimized for the modified process. At 35% solids content, the modified process had 80% lower slurry viscosity, 24% lower peak glucose concentration, 7.5% higher final ethanol concentration, and 51% higher fermentation rate compared with the conventional dry‐grind process. At 40% solids content, the modified process had lower viscosities, lower peak and residual glucose concentrations, and higher ethanol concentrations than the conventional process; however, the results were in contrast to those for 35% solids content. At 40% solids content, SSF did not run to completion for conventional or modified processes, and more than 2.5% w/v of residual glucose was left in the fermentation broth. Final ethanol concentration achieved with the modified process at 40% solids content was 19.5% v/v, similar to the ethanol concentration achieved with the modified process at 35% solids content. At 35% slurry solids content, a GSHE level of 1.25 μL/g db of corn and a GA level of 0.25 μL/g db of corn were selected as optimum enzyme doses for the modified process.     

Dry‐Grind Processing of Corn with Endogenous Liquefaction Enzymes

An amylase corn has been developed that produces an α‐amylase enzyme that is activated in the presence of water at elevated temperatures (>70°C). Amylase corn in the dry‐grind process was evaluated and compared with the performance of exogenous amylases used in dry‐grind processing. Amylase corn (1–10% by weight) was added to dent corn (of the same genetic background as the amylase corn) as treatments and resulting samples were evaluated for dry‐grind ethanol fermentation using 150‐g and 3‐kg laboratory procedures. Ethanol concentrations during fermentation were compared with the control treatment (0% amylase corn addition or 100% dent corn) which was processed with a conventional amount of exogenous α‐amylase enzymes used in the dry‐grind corn process. The 1% amylase corn treatment (adding 1% amylase corn to dent corn) was sufficient to liquefy starch into dextrins. Following fermentation, ethanol concentrations from the 1% amylase corn treatment were similar to that of the control. Peak and breakdown viscosities of liquefied slurries for all amylase corn treatments were significantly higher than the control treatment. In contrast, final viscosities of liquefied slurries for all amylase corn treatments were lower than those of the control. Protein, fat, ash, and crude fiber contents of DDGS samples from the 3% amylase corn treatment and control were similar.     

水媒法提取食用油技术研究进展

该文提出了水媒法提油技术的概念。水媒法提油技术是指以水为主要媒介的提油技术(水媒法),可辅以或不辅以与水互溶的可食用物质(例如乙醇)、食品级酶、超声波、微波等处理以破坏油料细胞壁和/或破乳的食用油提取方法。水媒法的提油过程主要分为分成破碎、提取、破乳和分离四个阶段。水媒法发展至今60余年,已全面发展了水代法、水酶法和乙醇水提法等多种制油技术。目前水媒法的产业化还存在一些问题,如油料的预处理、酶的种类与成本等。但是,水媒法绿色健康、符合可持续发展要求,随着工艺设备、分离技术等的发展,该技术将更加完善,成为未来未来取代传统工艺的新主流。该文将水媒法研究领域的成果进行归纳总结并为其未来发展提供理论参考。