Extraction of lipids from a specialist dairy stream

In this work a process for the extraction of neutral and complex lipids from the specialist dairy stream ‘beta-serum’ is described. Beta-serum is a proprietary product obtained from dairy streams containing greater than 60% fat that have been through phase inversion from an oil-in-water to a water-in-oil emulsion. It is an enriched source of milk fat globular membrane proteins and complex lipids, and is distinct from buttermilk in this respect. In the process described here a total lipid extract could be obtained from liquid beta-serum using a continuous near-critical dimethyl ether (DME) antisolvent fractionation process. Protein and water are precipitated from solution when mixed with the DME, whilst lipid and some water are extracted into the DME-rich phase. The extraction yield of lipids depended on the solids content of the feed and the feed to DME flow ratio, but did not depend on the lactose content. Lipids were also extractable from spray dried beta-serum powder, but only when the lactose content had been reduced below 45% by mass. A two step extraction process is described in which neutral lipids are extracted with supercritical CO₂, and then polar lipids using near-critical DME. The polar lipid extract was enriched in phospholipids (∼70% by mass), which included phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and sphingomyelin. Other complex lipid components extracted included gangliosides and cerebrosides. Unlike the antisolvent process, proteins were not denatured during either CO₂ or DME processing of the spray dried powders, and the de-fatted powders are therefore suitable for a range of functional foods. A polar lipid extract could also be produced from spray dried powder by extracting first with DME to obtain a mixed neutral/complex lipid extract, then re-extracting the lipid extract with CO₂ to remove neutral lipids.     

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.     

Subcritical co‐solvents extraction of lipid from wet microalgae pastes of Nannochloropsis sp.

In this paper subcritical co‐solvents extraction (SCE) of algal lipid from wet pastes of Nannochloropsis sp. is examined. The influences of five operating parameters including the ratio between ethanol to hexane, the ratio of mixed solvents to algal biomass (dry weight), extraction temperature, pressure, and time were investigated. The determined optimum extraction conditions were 3:1 (hexane to ethanol ratio), 10:1 ratio (co‐solvents to microalgae (dry weight) ratio), 90°C, 1.4 MPa, and 50 min, which could produce 88% recovery rate of the total lipids. In addition, electron micrographs of transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were conducted to show that the algal cell presented shrunken, collapsed with some wrinkles and microholes after SCE extraction. The main composition of total lipids extracted under the optimum conditions was TAG which represented more than 80%. And the fatty acid profile of triglycerides revealed that C16:0 (35.67 ± 0.2%), C18:1 (26.84 ± 0.044%) and C16:1 (25.96 ± 0.011%) were dominant. Practical applications: The reported method could save energy consumption significantly through avoiding deep dewatering (for example drying). The composition of the extracted lipid is suitable for the production of high quality biodiesel.     

Extraction of Lipids from Aqueous Protein‐Rich Streams using Near‐Critical Dimethylether

An extraction process for obtaining lipids from aqueous protein/lipid mixtures has been developed using near‐critical dimethylether (DME) as the solvent. The process has been demonstrated on fresh and reconstituted egg yolks, and on selected dairy by‐product streams. The lipid yield is dependent on the processing temperature, solids content and feed ratio of solvent to liquid. The extent of protein denaturation is also dependent on these parameters. The phase equilibria of DME‐water‐phospholipid mixtures are presented with modeling of the DME‐water system using the Peng‐Robinson equation of state.     

The lipids of corn germ and endosperm

Mature kernels of an inbred corn were hand dissected into germ and endosperm fractions. Among various solvents tested, boiling, water-saturatedn-butanol extracted the most lipid from endosperm, and it was used as t h e extracting solvent for both germ and endosperm. The germ contained 78% of the total lipids and the endosperm 17%. The most striking differences in the fatty acid compositions of the triglycerides and polar lipids were higher levels of stearic and linolenic acids in the endosperm lipids. Although precautions were taken during extraction to inactivate lipases, immediately after harvest the free fatty acid level of the total lipids of the whole kernel was 6.5%. Ninety-five percent of the free fatty acids was in the endosperm fraction where the free fatty acids made up 36.5% of the total lipids. In germ, free fatty acids represented only 0.6% of the total lipids. The individual phospholipid and glycolipid classes of the endosperm and germ lipids were similar except for high levels of lyso compounds in the endosperm lipids. The higher levels of linolenic acid, free fatty acids and lyso lipids in endosperm may affect the keeping quality of the corn grain and of fractions milled from the endosperm. Presented at the AOCS meeting, St. Louis, May 1978.     

Effect of extraction methods on lipid yield and fatty acid composition of lipid classes containing γ-linolenic acid extracted from fungi

The effect of extraction procedures on the lipid yield and fatty acid composition of total lipid and main lipid structures (phospholipids, diacylglycerols, triacylglycerols, free fatty acids, and sterol esters) of fungal biomass (Mucor mucedo CCF-1384) containing γ-linolenic acid (GLA) was investigated. Seventeen extraction methods, divided into three groups, were tested: six with chloroform/methanol, five with hexane/alcohols, and six with common solvents or mixtures. The chloroform/methanol procedure (2∶1) was selected as standard, where lipid yield (TL/DCW, total lipid per dry cell weight) was 17.8%, considered to be 100% of lipids present. All chloroform/methanol extractions yielded more than 83% recorvey of lipids. Use of hexane/isopropanol solvent systems led to a maximum of 75% recovery. The best lipid yield was achieved by a two-step extraction with ethanol and hexane (120%). Extraction efficiency of the other solvent systems reached a maximum of 73%. Triacylglycerols were the main structures of lipid isolated; only methanol-extracted lipid contained 58.5% phospholipids. The fatty acid content of total recovered lipid was variable and depended on both the lipid class composition and the solvent system. GLA concentrations in total lipids isolated by hexane/alcohol procedures (7.3–10.7%) are comparable with classical chloroform/methanol systems (6.5–10.0%). The maximal GLA yield was obtained with chloroform/methanol/n-butanol/water/0.1 M ethylenediaminetetraacetic acid (EDTA) (2∶1∶1∶1∶0.1, by vol) and after two-step extraction with ethanol and hexane (14.3 and 13.7 g GLA/kg DCW, respectively). The highest GLA content was analyzed in the phospholipid fraction (16.1%) after using chloroform/methanol/n-butanol/water/0.1 M EDTA (2∶1∶1∶1∶0.1, by vol). Remarkably low concentrations of polyunsaturated fatty acids were determined in the free fatty acid fraction.     

Optimization of algal lipid extraction by mixture of ethyl acetate and ethanol via response surface methodology for biodiesel production

The effects of extraction time, extraction temperature, solvent to biomass ratio and solvent composition on lipid yield from lyophilized Chlorococcum sp. biomass using a mixture of ethyl acetate and ethanol (EAE), a new proposed solvent, were studied. Subsequently, the process conditions of extraction by EAE were optimized using Box-Behnken design (BBD). The results revealed that the extraction temperature had the greatest effect on lipid extraction efficiency, followed by volume ratio of ethyl acetate to ethanol (EA/E) and extraction time. The largest lipid extraction yield of 15.74% was obtained under the following extraction conditions: 40mL solvents per gram of biomass for 270 min with gentle stirring at 80 °C by EAE with an EA/E of 1.0. Furthermore, palmitic acid, stearic acid, oleic acid, and linoleic acid were the most abundant fatty acids in the lipids extracted, indicating the great potential of the proposed lipid extraction procedure for microalgae-based biodiesel production.     

A Fast and Green Extraction Method for Berry Seed Lipid Extraction Using CO2 Expanded Ethanol Combined with Sonication

In this work, a combination of sonication and carbon dioxide expanded liquid extraction (SA‐CXLE) is used for the first time for the extraction of lipids from berry seeds. Three solvents (ethanol, methanol and ethyl lactate) are screened for the best recoveries of total lipids. Ethanol is selected due to its efficiency, greenness and sustainability. The effects of operation variables including temperature, time and CO₂ molar fraction on extraction performance are examined using a response surface methodology. The optimum conditions were found at 0.37 molar fraction of CO₂ in ethanol, a temperature of 52 °C and an extraction time of 7 min for two cycles. The SA‐CXLE method extracts 323 ± 38 mg g^?1 seed compared to 194 ± 23 using a conventional solid‐liquid extraction. SA‐CXLE is successfully applied to profile lipids from gooseberry, blackcurrant, chokeberry, red currant, and rowanberry seeds. More than 17 lipid classes are characterized and identified. Gooseberry shows the highest amount of oil of 352 mg g^?1 seed. Practical Applications: The developed method provides a fast, efficient and simple approach to extract and profile lipids using a combination of green solvents. Total lipid content and fatty acids composition of the berry seeds are essential information in pharmaceutical, cosmetic, food and nutritional applications.     

Optimization of cell wall disruption and lipid extraction methods by combining different solvents from wet Chlorella vulgaris

Over the past few decades, microalgae have emerged as a promising option for making lipid-based bioactive compounds. The purpose of this study was to extract lipids from Chlorella vulgaris. In this study, different cell wall disruption methods such as microwave, liquid nitrogen, ultrasound (US), and bead mill were compared. We selected and optimized two systems based on their efficacy in disrupting cell walls—US and beads mill. Based on the dry weight of C. vulgaris biomass, the maximum lipid extraction by the US was 17.1% and by bead mill was 15.2%. Following cell wall disruption of C. vulgaris, chloroform–methanol (2:1) solvent combination achieved high lipid extraction. However, the hexane–ethanol (1:1) solvent combination was chosen because of its lower toxicity. Specifically, the effect of the solvent-to-biomass ratio, the temperature, and the extraction time was investigated. The results indicated that the chloroform–methanol solvent combination yielded optimal results at 8 ml/g solvents to biomass, 45°C, and 60 min and that the hexane–ethanol combination yielded optimal results at 6 ml/g, 35°C, and 73 min, respectively. The highest amount of lipids was obtained from C. vulgaris with 87.6% moisture content. As a cell wall disruption method, the US obtained 20.4% and 16.4% with a combination of chloroform–methanol solvents and hexane–ethanol, respectively. Additionally, bead milling resulted in the highest extraction yield of 17.6% for chloroform–methanol and 13.9% for hexane–ethanol. Based on the results of cell wall disruption, the US method is the most efficient cell wall disruption method in terms of lipid extraction efficiency.     

Development of a new procedure for lipid extraction from Hevea brasiliensis natural rubber

Non‐isoprene components and especially lipids have been reported to be involved in some key properties of natural rubber. Unfortunately, these results are hardly comparable due to different extraction methods. This work aimed to optimize lipid extraction from natural rubber either in the liquid state (latex) or in the dry state (unsmoked sheets). Extraction of unsmoked sheets from the RRIM 600 clone was carried out with different combinations of organic solvents (chloroform/methanol and hexane/isopropanol mixes). Chloroform/methanol (2 : 1 vol/vol) was found to be the most suitable for lipid extraction from unsmoked sheet rubber. The lipid extraction yield was improved by increasing the exchange surfaces by grinding rubber under liquid nitrogen and extracting the ground rubber for 6 h at room temperature, leading to 1.82% lipid extraction yield (versus dry rubber). Concerning latex extraction, the problem of lipid entrapment in the coagulum from immediate coagulation of latex in the solvent was solved by preliminary two times dilution of latex, giving a 3.24% extract (versus dry rubber) containing a minimum quantity of contaminating polyisoprene. Concerning the nature of lipids, dilution increased mainly neutral lipid extraction, which may suggest that neutral lipids were those entrapped by coagulation.     

Extraction of rice brain oil using supercritical carbon dioxide and propane

Extraction of rice bran lipids was performed using supercritical carbon dioxide (SC−CO₂) and liquid propane. To provide a basis for extraction efficiency, accelerated solvent extraction with hexane was performed at 100°C and 10.34 MPa. Extraction pressure was varied for propane and SC−CO₂ extractions. Also, the role of temperature in SC−CO₂ extraction efficiency was investigated at 45,65, and 85°C. For the SC−CO₂ experiments, extraction efficiencies were proportional to pressure and inversely proportional to temperature, and the maximal yield of oil achieved using SC−CO₂ was 0.222±0.013 kg of oil extracted per kg of rice bran for conditions of 45°C and 35 MPa. The maximal yield achieved with propane was 0.224±0.016 kg of oil per kg of rice bran at 0.76 MPa and ambient temperature. The maximum extraction efficiencies of both SC−CO₂ and propane were found to be significantly different from the hexane extraction baseline yield, which was 0.261±0.005 kg oil extracted per kg of rice bran. A simulated economic analysis was performed on the possibility of using SC−CO₂ and propane extraction technologies to remove oil from rice bran generated in Mississippi. Although the economic analysis was based on the maximal extraction efficiency for each technology, neither process resulted in a positive rate of return on investment.     

Extraction of Phospholipids from Structured Dry Egg Yolk

Chicken egg yolk is a concentrated source of phospholipids (PL). Extracting egg PL with high efficiency is vital to the availability and economics of this high-valued lipid product. In this study, two types of structured dry egg yolk materials, yolk flakes and pellets, were prepared. Two commonly used solvents, hexane and ethanol, were tested on the extraction of total yolk lipids including the PL. The PL fraction was obtained by the conventional cold acetone precipitation. The drum-dried yolk flakes were shown to be an ideal starting material for total lipid and PL extraction. Anhydrous ethanol can extract almost all the neutral lipids and PL with little change to the individual components of the native PL. A PL product with a purity of more than 90 % and a yield of 99 % can be prepared using this method.     

Extraction of pomegranate seed oil using subcritical propane and supercritical carbon dioxide

Extraction of oil from pomegranate seeds as a waste product of the juice industry using supercritical carbon dioxide and subcritical propane was studied in this work. The influence of the main operating conditions of extraction, namely, the temperature and pressure of extraction on the oil extraction yield and the correspondent fatty acid profile were analyzed and reported here. Soxhlet extraction with n-hexane was done for comparison with supercritical extraction resulting in a maximum yield of oil of 22.31 wt %. Supercritical carbon dioxide and subcritical propane extracted up to 58.53% (corresponding to maximum yield of 13.06 wt %) and 76.73% (corresponding to maximum yield of 17.12 wt %) of the total amount of pomegranate seed oil as measured using Soxhlet extraction, respectively. Results indicated that the subcritical propane is a suitable and selective solvent for the extraction of the pomegranate seed oil in function of smaller times and pressures employed compared to carbon dioxide extraction. The fatty acid composition of the extracted oil showed the presence of fatty acids of C₁₆, C₁₈, C₂₀, C₂₂, and C₂₄ carbon chains. Punicic (C_18.3) was the major fatty acids and comprise up to 70% of the total fatty acid content of the extracted oil among all samples. Finally, Lack??s plug flow model as developed by Sovova was applied to both extraction systems and a good agreement with the experimental results was obtained.     

Extraction of sesame seed (Sesamun indicum L.) oil using compressed propane and supercritical carbon dioxide

This work is aimed to investigate the extraction of sesame seed (Sesamun indicum L.) oil using supercritical carbon dioxide and compressed propane as solvents. The extractions were performed in a laboratory scale unit in a temperature and pressure range of 313-333 K and 19-25 MPa for carbon dioxide and 303-333 K and 8-12 MPa for propane extractions, respectively. A 2² factorial experimental design with three replicates of the central point was adopted to organize the data collection for both solvents. The results indicated that solvent and density were important variables for the CO₂ extraction, while temperature is the most important variable for the extraction yield with propane. The extraction with propane was much faster than that with carbon dioxide due to the fact that propane is a better solvent for vegetable oils compared to carbon dioxide. On the other hand, characteristics of extracted oil, its oxidative stability determined by DSC and chemical profile of constituent fatty acids determined by gas chromatography, were similar to both solvents. The mathematical modeling of the extraction kinetics using a second order kinetic presented good results for the extraction with both solvents.     

Lipid extraction from the microalga Phaeodactylum tricornutum

Ethanol was used for the extraction and purification of lipids from the biomass of the microalga Phaeodactylum tricornutum. This microalga is an oil‐rich substrate with a high proportion of eicosapentaenoic acid (EPA). The process consisted of two steps. First, ethanol (96% vol/vol) was used to extract the lipids from the lyophilized biomass. Second, a biphasic system was formed by adding water and hexane to the extracted crude oil. In this way, most of the lipids were transferred to the hexanic phase while most impurities remained in the hydroalcoholic phase. The first step was carried out by two consecutive extractions at room temperature, each with 5 mL ethanol per gram of biomass, for 10 and 1.25 h, respectively. Under these conditions, over 90% of the saponifiable lipids in the biomass were extracted. In the second step, the percentage of water in the hydroalcoholic phase, the hexane/hydroalcoholic phase ratio and the number of extraction steps were optimized. A water content of 40% vol/vol in the hydroalcoholic phase provided the highest lipid recovery. A recovery yield of 80% was obtained by four consecutive extractions with a hexane/hydroalcoholic phase ratio of 0.2 (vol/vol). Equilibrium distribution data of the lipids between the hydroethanolic and the hexanic phases were also obtained in order to predict the lipid recovery yield of the extraction. This process is an alternative to the traditional methods of lipid extraction, which uses less toxic solvents and reduces the total amount of solvents used.     

Comparison of the lipid composition ofCulex quinquefasciatus andCulex tritaeniorhynchus cells obtained from the logarithmic and stationary phases of growth

Culex quinquefasciatus andCulex tritaeniorhynchus cells were grown in spinner culture medium. The cells were harvested at late logarithmic and stationary phases of growth. The total lipid, total neutral lipid, and total phospholipid contents of the cells were analyzed to determine changes that occurred in the fatty acid profiles of the lipids with aging and between species. There was an increase in the amount of total neutral lipids with a corresponding decrease in amount of total phospholipid from logarithmic to stationary phases of growth of theCulex quinquefasciatus cells. Chain elongation and/or desaturation of acids occurred with aging of cells. The fatty acids of the phospholipids had a longer average chain length than the neutral lipids.     

亚临界流体萃取贡菊净油的工艺优化

为了解决传统溶剂萃取贡菊净油过程中溶剂残留和香味物质损失的问题,通过二甲醚亚临界萃取贡菊的单因素实验及正交实验,研究了萃取压力、萃取温度、萃取时间及液固比对萃取提取率的影响,确定了二甲醚亚临界萃取提取贡菊净油的最佳工艺条件。结果表明,各影响因素对贡菊净油提取率作用的大小依次为:萃取压力液固比萃取温度萃取时间。最佳工艺参数为:萃取压力0.5MPa、萃取温度20℃、萃取时间3 h、液固比7∶1 m L/g,贡菊净油提取率平均值为4.19%。该净油可提升烟草香气丰富性,能赋予卷烟产品独特的风格特征,明显提升卷烟的抽吸品质。优化得到的工艺可提高贡菊净油的得率,工艺可行稳定。     

Structural and functional role of lipids in yeast and mycelial forms ofCandida albicans

The levels of total lipids, sterols and phospholipids were found to be significantly higher in the mycelial form (log phase) ofCandida albicans than in the yeast form. Increased phospholipid levels in the mycelial form were due to higher levels of phosphatidylcholine, phosphatidylserine and phosphatidylinositol. Analyses of fatty acid composition also revealed higher levels of myristic acid (40%) in the yeast form, resulting in higher levels of saturated lipids than in the mycelial form. The changes in the lipid composition were also manifested in altered thermotropic phase behavior as gel-to-liquid crystalline phase transitions were observed at 36 and 27°C for the lipids of the yeast and mycelial forms, respectively. These changes coincided with higher uptake rate, i.e., Km and Vmax values, for the transport of L-proline and with a higher sensitivity of the mycelial form against antifungal drugs.     

A rapid method for extraction of total lipids from whey protein concentrates and separation of lipid classes with solid phase extraction

A modified procedure for extraction of total lipids from whey protein concentrates was developed such that stable emulsion with extracting solvents was avoided and the solvent system remained monophasic. Nonlipid contaminants from the extract were removed using gel filtration instead of traditional aqueous washing to prevent any loss of polar lipids. The extraction of total lipids by the modified procedure was complete and comparable with a reference procedure. Traditional thin-layer chromatography is tedious and more qualitative than quantitative for lipid class separation. Total lipids were further separated into free fatty acids, phospholipids, cholesterol ester, triacylglycerol, cholesterol, diacylglycerol, and monoacylglycerol, using modified solid phase extraction procedure. Columns with 2 g amino propyl packing allowed separation of up to 80 mg of total lipids into lipid classes gravimetrically. The values for anhydrous milk fat for all lipid classes agreed with those in the literature. Separation of total lipids into lipid classes with solid phase extraction is easy, quantitative, and can also be performed on a preparative scale.     

Comparison of supercritical fluid and solvent extraction methods in extracting γ-oryzanol from rice bran

Organic solvents were compared with supercritical CO₂ relative to efficiency for extracting lipid and γ-oryzanol from rice bran. A solvent mixture with 50% hexane and 50% isopropanol (vol/vol) at a temperature of 60°C for 45–60 min produced the highest yield (1.68 mg/g of rice bran) of γ-oryzanol among organic solvents tested. The yield of γ-oryzanol without saponification was approximately two times higher (P<0.05) than that with saponification during solvent extraction. However, the yield (5.39 mg/g of rice bran) of γ-oryzanol in supercritical fluid extraction under a temperature of 50°C, pressure of 68,901 kPa (680 atm), and time of 25 min was approximately four times higher than the highest yield of solvent extraction. Also, a high concentration of γ-oryzanol in extract (50–80%) was obtained by collecting the extract after 15–20 min of extraction under optimized conditions.