Concentration of phytosterols for analysis by supercritical fluid extraction

Fractionation of sterols from plant lipid mixtures was accomplished using a multistep supercritical fluid extraction (SFE) procedure. Samples of seed oils, margarine, corn germ oil, and corn fiber oil were extracted to yield enriched phytosterol fractions. Supercritical fluid chromatography (SFC) was utilized to separate and determine the concentration of the plant sterols in the extracts from the various samples. The sterol concentration in the original samples varied from 2.2 mg/g in soybean oil to 13.2 mg/g in oil extracted from corn fiber. After the SFE-based fractionation of the samples, the sterol concentration was increased to 64.4 mg/g in the extract from soybean oil and 166.2 mg/g in the extract from corn fiber oil. Oil extracted from corn bran, which measured 8.6 mg/g in the original oil, increased to 322.2 mg/g using the fractionation process. The benign conditions utilized by SFE and SFC proved to be effective for the analyses of these compounds without inducing degradation of the analytes.     

Antioxidant activity of phytosterols, oryzanol, and other phytosterol conjugates

Antioxidant activity of phytosterols, oryzanol, ferulic acid ester of sterols, corn fiber oil, and rice bran oil was investigated. Commercial soybean oil and distilled soybean oil FAME were used as substrates for both oxidative stability determination and viscosity analysis after the oil was oxidized. At low concentration, these materials did not improve the oxidative stability of the oil substrates, although the viscosity tended to be reduced slightly. The antipolymerization activity of steryl ferulate was higher at higher concentration than at lower concentration, and steryl ferulate was more effective than oryzanol. Rice bran oil showed very good antioxidant and antipolymerization activities.     

The Composition of Crude Corn Oil Recovered after Fermentation via Centrifugation from a Commercial Dry Grind Ethanol Process

A study was conducted to examine the chemical composition of corn oil obtained after fermentation of corn to make fuel ethanol via centrifugation and compare its composition to that of corn germ oil (commercial corn oil) and experimental corn oils. The levels of free fatty acids in the post fermentation corn oil were high (11–16%), as previously reported. The levels of free phytosterols and hydroxycinnamate steryl esters (similar to oryzanol in rice bran oil) were higher than those of corn germ oil and were comparable to those of ethanol-extracted corn kernel oil. The levels of tocopherols were lower in post-fermentation oil than in either corn germ oil or ethanol extracted corn kernel oil. The levels of lutein and zeaxanthin in post-fermentation were much higher than those in corn germ oil and were comparable to those in ethanol-extracted corn kernel oil. Overall, exposure to all upstream processes of a fuel ethanol plant, including high-temperature liquefaction, saccharification and fermentation appeared to have the most notable effect on tocopherols, but it had little effect on the levels of free phytosterols, hydroxycinnamate steryl esters, lutein and zeaxanthin. It may be desirable to recover these valuable functional lipids prior to using the post-fermentation corn oil for industrial applications such as making biodiesel if a cost-effective recovery process can be developed.     

Sterol content of some plant oils; Further observations on fast-reacting sterols

Total and free sterols were measured by a modified Sperry-Webb procedure in raw and refined corn, cottonseed, safflower seed, linseed, soybean and wheat germ oils. Wheat germ and safflower seed oil sterols were relatively rich in fastreacting sterols, which predominated in the sterol ester fraction. Photometric constants following color reaction were obtained for cholesterol and plant sterols and applied to a procedure for analysis of cholesterol and plant sterols in mixtures thereof.     

Fatty Acid,Phytosterol, and Polyamine Conjugate Profiles of Edible Oils Extracted from Corn Germ,Corn Fiber,and Corn Kernels

This study compared the profiles of fatty acids, phytosterols, and polyamine conjugates in conventional commercial corn oil extracted from corn germ and in two “new-generation” corn oils: hexane-extracted corn fiber oil and ethanol-extracted corn kernel oil. The fatty acid compositions of all three corn oils were very similar and were unaffected by degumming, refining, bleaching, and deodorization. The levels of total phytosterols in crude corn fiber oil were about tenfold higher than those in commercial corn oil, and their levels in crude corn kernel oil were more than twofold higher than in conventional corn oil. When corn kernel oil was subjected to conventional degumming, refining, bleaching, and deodorization, about half of the phytosterols was removed, whereas when corn fiber oil was subjected to a gentle form of degumming, refining, bleaching, and deodorization, only about 10% of the phytosterols was removed. Finally, when the levels of polyamine conjugates (diferuloylputrescine and p-coumaroyl feruloylputrescine) were examined in these corn oils, they were only detected in the ethanol-extracted crude corn kernel oil, confirming earlier reports that they were not extracted by hexane, and providing new information that they could be removed from ethanol-extracted corn kernel oil by conventional degumming, refining, bleaching, and deodorizing.     

Lipid Composition of Wheat Germ and Wheat Germ Oil

Information available in the literature concerning the composition of lipids in wheat germ and in wheat germ oil is critically reviewed. After a brief introduction to the botanical and technological aspects of wheat germ, the yield of oil and its physico-chemical properties are described followed by the composition of fatty acids, acyl lipids and non-saponifiable components. The importance of distinguishing between dissected germ and commercial wheat germ and between germ oil and germ lipids is emphasised. The triglycerides account for the major part of the fatty acids, of which linoleic acid is the principal component, and the content of free fatty acids depends on the rancidity of the germ and also on possible post-extraction processing in the case of commercial oil. Polar lipids consist mainly of phospholipids and available information suggests that glycolipids are present only at very low concentration and that galactosyl glycerides may be absent from the embryo of the quiescent wheat grain. Most reports of tocopherol composition concern wheat germ oil and there is less information about the tocopherols of dissected wheat germ. α- and β-tocopherols are found in wheat germ but tocotrienols are probably absent from dissected germ and only occur in commercial germ as a result of bran and endosperm contamination. Wheat germ oil exhibits a range of sterols. 4-methyl sterols and triterpenoid alcohols, and β-sitosterol and campesterol are the major components. The hydrocarbon composition of wheat germ oil has been reported but the significance of the results is uncertain. Flavonoid pigments, xanthophyll and xanthophyll esters have been shown to be present in wheat germ. Most of the studies of the non-saponifiable fraction were based on germ oil and commercial wheat germ and little is known of the nature of this fraction in dissected wheat germ.     

Pearling barley and rye to produce phytosterol-rich fractions

Because of the positive health effects of phytosterols, phytosterol-enriched foods and foods containing elevated levels of natural phytosterols are being developed. Phytosterol contents in cereals are moderate, whereas their levels in the outer layers of the kernels are higher. The phytosterols in cereals are currently underutilized; thus, there is a need to create or identify processing fractions that are enriched in phytosterols. In this study, pearling of hulless barley and rye was investigated as a potential process to make fractions with higher levels of phytosterols. The grains were pearled with a laboratory-scale pearler to produce pearling fines and pearled grains. Lipids were extracted by accelerated solvent extraction, and nonpolar lipids were analyzed by normal-phase HPLC with ELSD and UV detection. Total sterol analyses were performed by GC. After a 90-s pearling, the amounts of pearling fines from hulless barley and rye were 14.6 and 20.1%, respectively, of the original kernel weights. During pearling, higher levels of phytosterols and other lipids were fractionated into the fines. The contents of free sterols and sterols esterified with FA in the fines were at least double those in the whole grains. Pearling fines of hulless barley and rye contained >2mg/g phytosterol compounds, which makes them a good source of phytosterols and thus valuable raw materials for health-promoting foods.     

Preparative-scale supercritical fluid extraction/supercritical fluid chromatography of corn bran

A preparative-scale supercritical fluid extraction/supercritical fluid chromatography (SFE/SFC) procedure has been developed for the removal of oil from corn bran to obtain fractions enriched with free sterols and ferulate-phytosterol esters (FPE). Operational parameters from an analytical-scale supercritical fluid fractionation technique were translated to and optimized on a home-built, preparatory-scale SFE/SFC apparatus. SFE was performed at 34.5 MPa and 40°C using supercritical carbon dioxide. These conditions did not result in exhaustive extraction of the corn bran, but yielded about 96% of the available oil. SFC was conducted in three steps, followed by reconditioning of the sorbent bed. Preparative-scale SFE/SFC of corn bran produced a fraction enriched greater than fourfold in free sterols and 10-fold in FPE, suggesting that such a scheme could be used industrially to produce a functional food ingredient.     

The composition of corn oil obtained by the alcohol extraction of ground corn

All commercial corn oil is obtained by the hexane extraction of corn germ. The chemical composition of commercial corn oil has been well characterized. This study was under-taken to quantitatively evaluate the lipid composition of corn oil obtained by the ethanol extraction of ground, whole corn kernels. When corn oil was obtained by extracting ground corn kernels (ground corn) with polar or nonpolar solvents, the resulting corn oil contained much higher levels of hydroxycinnamate steryl esters (≈0.3%) than those found in commercial hexane-extracted corn (germ) oil (≈0.02%). The levels of valuable tocopherols and tocotrienols were also significantly higher in kernel oil than in traditional corn germ oil. We previously reported that when corn oil was obtained by extracting corn kernels with polar solvents, the oil contained two polyamine conjugates, diferuloylputrescine and p-coumaroyl feruloylputrescine. In the current study, when ground corn was extracted with ethanol, the resulting corn oil contained about 0.5% diferuloylputrescine and about 0.2% p-coumaroyl feruloylputrescine. This is the first study to quantify these unique compounds in corn oil extracted by new techniques. This compositional information is important because this new oil is being considered for human food use.     

Characterization of Oil Precipitate and Oil Extracted from Condensed Corn Distillers Solubles

Oil extracted from condensed corn distillers solubles (CCDS) can form a semi-solid and waxy precipitate at the bottom of containers during storage. CCDS is a good source to recover oil, and such oil can be converted to biodiesel. Precipitate formation in the extracted oil is mainly a physical stability problem, but it may become a performance problem for biodiesel. The objective of the present work was to determine the composition of the CCDS oil precipitate and also determine if valuable phytosterols were present in high concentration. The free fatty acid (FFA) content was very high, 35.7%, and fatty acid composition of the FFA fraction was predominantly palmitic acid, 70.3%. The solid appearance was mainly due to a high percentage of high-melting point free saturated fatty acid. The total unsaponifiable matter was 2.0%, and total phytosterol content was 8.6 mg/g of CCDS oil precipitate. Therefore, CCDS oil precipitate is a not an enriched source of phytosterols compared to total sterols present in crude corn oil (15.6 mg/g oil). The wax content was high, 2.5 mg/g of CCDS oil precipitate compared to 0.5 mg/g of crude corn oil. CCDS oil that is uncentrifugable but polar solvent extractable (trapped oil fraction) was also characterized and found to contain more polar lipids than those in the free oil fraction (centrifugable oil).     

Diferuloylputrescine and p-coumaroyl-feruloylputrescine, abundant polyamine conjugates in lipid extracts of maize kernels

Extraction of corn bran or corn fiber with polar solvents such as methylene chloride, ethanol or chloroform/methanol yielded common lipids and two unknown high-performance liquid chromatography (HPLC) peaks, each with an ultraviolet absorbance maximum at 320 nm. HPLC-mass spectrometry revealed that the unknowns were diferuloylputrescine (DFP) and p-coumaroyl-feruloylputrescine (CFP). When compared to extracts of corn fiber (a pericarp-enriched fraction from the wet milling of corn), comparable extracts of corn bran (a pericarp- enriched fraction from the dry milling of corn) yielded three- to eightfold higher levels of DFP and CFP. Extraction of corn bran or fiber with an accelerated solvent extractor revealed that elevated temperatures greatly enhanced the extraction of DFP and CFP by methylene chloride and ethanol. “Corn bran oil,” prepared by extraction of corn bran with hot methylene chloride, contained 14 wt% DFP and 3 wt% CFP. However, when hexane was used as a solvent, accelerated solvent extraction of the corn bran or fiber did not extract any DFP or CFP. Extraction of wheat bran or psyllium hulls with hot methylene chloride did not yield any detectable DFP or CFP. Because it has been suggested that polyamine conjugates such as DFP and CFP may function as natural pesticides, a rapid method was developed to purify them so that their biological activity could be evaluated.     

Sterol additives as polymerization inhibitors for frying oils

Addition of certain vegetable oil unsaponifiables to safflower oil protects it from oxidative polymerization during heating at frying temperature. The unsaponifiables isolated from olive, corn, wheat germ andVernonia anthelmintica oils were found to be effective. The fraction responsible for this effect is largely sterol in nature. Although the common plant sterols show no antioxidant activity, the 4-α-methyl sterols function well. The sterols fromVernonia oil, which contain no 4-α-methyl group, are also active. It appears that an isofucosterol side chain may be the structural feature required to obtain this protective effect.     

Analysis of sterols from various food matrices

Two methods suitable for routine phytostanol/phytosterol analysis of various sterol‐enriched food matrices and phytostanyl/phytosteryl fatty acid ester ingredients are introduced. A method based on hot saponification of a sample with ethanolic potassium hydroxide in the presence of an internal standard (5β‐cholestan‐3α‐ol) is adequate for most matrices, such as spread, milk and yoghurt. Some matrices, like pasta, require acid hydrolysis in order to release matrix‐incorporated bound sterols or sterols from steryl glycosides before the saponification step. After saponification, the unsaponifiable material containing phytostanols and phytosterols is extracted into an organic solvent (e.g. heptane), followed by evaporation of the solvent to dryness. Sterols are separated as their trimethylsilyl ether derivatives with a gas‐liquid chromatograph (GC), on a column coated with 5% phenyl/95% dimethylpolysiloxane, and detected with a flame ionization detector. The GC conditions applied provide efficient separation of the most abundant phytostanols/phytosterols in 15 min, a wide linear range of stanols/sterols without the need of defining sterol response factors. The methods are repeatable and accurate, as shown with standard addition trials. These methods were applied to determine phytostanol/phytosterol contents of several sterol‐enriched functional food products, and the analyzed amounts were in good accordance with the information provided on the packaging labels.     

米糠精制产品的应用

对米糠综合利用的途径进行了详细论述；并总结厂各种米糠精制产品在日用化工、医药工业、食品工业、精细化工领域的具体用途，包括米糠油的浸提技术，米糠油作为营养保健食品的开发利用，米糠油作为油脂化工原材料的深加工；米糠油精炼皂脚中提取游离脂肪酸及脂肪酸衍生物的制备；米糠脱水、脱臭、脱色的小皂化物提取谷甾醇、生育酚、谷维素的方法；米糠脱蜡副产物制备糠蜡和二十烷醇的利用及米糠饼(粕)提取植酸钙、植酸和肌醇的利用途径，最后提出了大力发展我国米糠产业的市场前景。     

Phytosterol oxidation products (POP) in foods with added phytosterols and estimation of their daily intake: A literature review

To evaluate the content of phytosterol oxidation products (POP) of foods with added phytosterols, in total 14 studies measuring POP contents of foods with added phytosterols were systematically reviewed. In non‐heated or stored foods, POP contents were low, ranging from (medians) 0.03–3.6 mg/100 g with corresponding oxidation rates of phytosterols (ORP) of 0.03–0.06%. In fat‐based foods with 8% of added free plant sterols (FPS), plant sterol esters (PSE) or plant stanol esters (PAE) pan‐fried at 160–200°C for 5–10 min, median POP contents were 72.0, 38.1, and 4.9 mg/100 g, respectively, with a median ORP of 0.90, 0.48, and 0.06%. Hence resistance to thermal oxidation was in the order of PAE > PSE > FPS. POP formation was highest in enriched butter followed by margarine and rapeseed oil. In margarines with 7.5–10.5% added PSE oven‐heated at 140–200°C for 5–30 min, median POP content was 0.3 mg/100 g. Further heating under same temperature conditions but for 60–120 min markedly increased POP formation to 384.3 mg/100 g. Estimated daily upper POP intake was 47.7 mg/d (equivalent to 0.69 mg/kg BW/d) for foods with added PSE and 78.3 mg/d (equivalent to 1.12 mg/kg BW/d) for foods with added FPS as calculated by multiplying the advised upper daily phytosterol intake of 3 g/d with the 90% quantile values of ORP. In conclusion, heating temperature and time, chemical form of phytosterols added and the food matrix are determinants of POP formation in foods with added phytosterols, leading to an increase in POP contents. Practical applications: Phytosterol oxidation products (POP) are formed in foods containing phytosterols especially when exposed to heat treatment. This review summarising POP contents in foods with added phytosterols in their free and esterified forms reveals that heating temperature and time, the chemical form of phytosterols added and the food matrix itself are determinants of POP formation with heating temperature and time having the biggest impact. The estimated upper daily intakes of POP is 78.3 mg/d for fat‐based products with added free plant sterols and 47.7 mg/d for fat‐based products with added plant sterol esters. Phytosterols in foods are susceptible to oxidation to form phytosterol oxidation products (POP). This review summarizes literature data regarding POP contents of foods with added phytosterols that were exposed to storage and heat treatments.     

Pressurized liquid extraction of polar and nonpolar lipids in corn and oats with hexane,methylene chloride,isopropanol, and ethanol

Samples of freshly ground corn kernels and freshly ground rolled oats were extracted via pressurized liquid extraction (accelerated solvent extraction) using four different organic solvents [hexane, methylene chloride (also known as dichloromethane), isopropanol, and ethanol] at two temperatures (40 and 100°C). Lipid yields varied from 2.9 to 5.9 wt% for ground corn and from 5.5 to 6.7 wt% for ground oats. With ground corn, more lipid was extracted as solvent polarity was increased, and for each individual solvent, more lipid was extracted at 100°C than at 40°C. With ground oats, the same temperature effects was observed, but the solvent polarity effect was more complex. For both corn and oats, methylene chloride extracted the highest levels of each of the nonpolar lipid classes. In general, for both corn and oats, icnreasing solvent polarity resulted in increasing yields of polar lipids, and for each solvent, more of each lipid class was extracted at 100°C, than at 40°C. Among the lipids in corn extracts, the phytosterols may be the most valuable, and total phytosterols ranged from about 0.6 wt% in the hot ethanol extracts to about 2.1 wt% in the hot hexane and methylene chloride extracts. Total phytosterols in all oat extracts were about 0.1 wt%. Digalactosyldiacylglycerol was the most abundant polar lipid in the oat extracts; its levels ranged from 1.6 wt% in the cold hexane extracts to 4.3 wt% in the hot ethanol extracts.     

Extraction and Identification of Phytosterols in Manketti (Schinziophyton rautanenii) Nut Oil

Phytosterols of manketti (Schinziophyton rautanenii) nut oil extracted by Soxhlet, mechanical shaking using hexane, screw press and supercritical carbon dioxide, were analyzed by gas chromatography with flame ionization detection and identified by gas chromatography–mass spectrometry. The presence of several phytosterols (campesterol, stigmasterol, β‐sitosterol, Δ⁵‐avenasterol, 22‐dihydrospinasterol and Δ⁷‐avenasterol) previously reported in manketti oil, was confirmed. In addition, another fourteen phytosterols (lanosterol, Δ^5,23‐stigmastadienol, Δ⁷‐campesterol, clerosterol, obtusifoliol, Δ^5,24(25)‐stigmastadienol, α‐amyrin, gramisterol, cycloeucalenol, cycloartenol, stigmasta‐8,24‐dienol‐3‐β‐ol, 28‐methylobtusifoliol, 24‐methylenecycloartenol and citrostadienol) were identified. The phytosterols, β‐sitosterol, Δ⁵‐avenasterol and campesterol, had the highest concentrations in oils extracted by all the methods, whereas stigmasterol and cycloartenol were abundant in oils extracted by mechanical shaking and supercritical carbon dioxide. Total phytosterols and the quantities of individual phytosterols differed significantly (p ≤ 0.05) in oils from the four extraction methods. Mechanical shaking extracted the highest levels of total sterols (22,100 mg/100 g oil), followed by supercritical fluid extraction (9,550 mg/100 g oil). Screw press and Soxhlet extracted oils contained the lowest levels of total sterols, 3,810 mg/100 g oil and 3,350 mg/100 g oil, respectively.     

Contents of Carotenoids, Tocopherols and Sterols in Acacia cyanophylla Seed Oils

Seeds from 12 Acacia cyanophylla ecotypes, harvested in Tunisia, were examined for their seed oil contents of carotenoids, tocopherols and phytosterols. The average carotenoid content (lutein and zeaxanthin) was ca. 102 mg kg^?1 of total extracted lipids. Lutein (ca. 97 mg kg^?1 of total extracted lipids) was usually more abundant than zeaxanthin (ca. 5 mg kg^?1 of total extracted lipids). The mean total tocopherol content was ca. 704 mg kg^?1 of total extracted lipids. The main isomer was α‐tocopherol, with more than 75 % of total tocopherols (ca. 528 mg kg^?1 of total extracted lipids), followed by γ‐tocopherol (ca. 168 mg kg^?1 of total extracted lipids) and δ‐tocopherol (ca. 86 mg kg^?1 of total lipids). High levels of phytosterols (ca. 7.8 g kg^?1 of total extracted lipids) were detected, among which β‐sitosterol was the most abundant (47 %). All these results highlight the richness of carotenoids, tocopherols and sterols in A. cyanophylla seed oil, and imply that this species might constitute a potential resource for the development of functional foods.     

Triterpene alcohols and sterols of vegetable oils

Triterpene alcohols and sterols were separated by thin-layer chromatography and gas-liquid chromatography from the unsaponifiable fractions of the following 18 vegetable oils: linseed, peanut, olive, rice bran, palm kernel, corn, sesame, oiticica, palm, coconut, rapeseed, grape seed, sunflower, poppy seed, castor, tea seed, cocoa butter and soybean. Two triterpene alcohols, cycloartenol and 24-methylene cycloartanol, were found in all of the oils except soybean oil, which contained only cycloartenol. Triterpene alcohols such as α- and β-amyrin, euphorbol, butyrospermol and cyclolaudenol also were encountered occasionally. Three sterols, β-sitosterol, stigmasterol and campesterol were present in all of the oils. In addition a fourth sterol, not yet idenfified, was found in oils of palm, palm kernel and sunflower in varying amounts. This unknown sterol and brassicasterol were found in rapeseed oil in addition to the three sterols that were common to all of the oils studied. Experiment Station for Fats and Oils, National Center for Lipochemistry of National Research Council, Milan, Italy.     

Vorkommen bitterer Hydroxyfettsäuren in Hafer und Weizen

Occurrence of Bitter Hydroxy Fatty Acids in Oat and Wheat Water suspensions of oat, soft wheat, and durum wheat flours were incubated for 4.5 h at 38° C. Only the oat flour became intensely bitter. Freeze-dried samples of the incubated flours were analyzed for free hydroxy fatty acids with bitter taste. 750 μg/g L-OH (mixture of 9-hydroxy-trans, cis-10,12-octadecadienoic acid and 13-hydroxy-cis, trans-9,11-octadecadienoic acid) and 62 μg/g Tri-OH (mixture of 9,12,13-trihydroxy-trans-10-octadecenoic acid and 9,10,13-trihydroxy-trans-11-octadecenoic acid) were found in oat flour. Soft wheat flour contained the same amount of Tri-OH as oat flour but only 60 μg/g LOH were detected. Considerable less amounts of both L-OH and Tri-OH (20 μg/g and 15 μg/g respectively) were found in durum wheat flour. The bitter taste of oat flour which has been debittered by extraction of the lipids could be restored by addition of a mixture of 750 μg/g L-OH and 62 μg/g Tri-OH.