Anhydrous Milk Fat Fractionation with Continuous Countercurrent Supercritical Carbon Dioxide

A continuous pilot-scale supercritical carbon dioxide system was designed and built for separation and fractionation up to 400 g/hr of anhydrous milk fat. This fat was separated into five fractions (S1-S5) in the pressure range 24.1–3.4 MPa at 40–75°C. The solvent/feed ratio was 62 with extraction yield 78 wt%. The short-chain (C4-C8) and medium-chain (C10-C12) fatty acids increased from S1S5. The long-chain and unsaturated fatty acids (C14-C18) decreased gradually from S1-S5. The triglycerides followed the same trend as fatty acids. Cholestrol content decreased by 51% and β-carotene concentration increased by 145% in the raffinate (S1).     

Phase Equilibria and Distribution Coefficients of δ-Lactones in Supercritical Carbon Dioxide

A static method was used to measure the solubilities and distribution coefficients of δ-lactones (pure), inherent (natural levels) δ-lactones of anhydrous milk fat (AMF) and fortified (added) δ-lactones in AMF in supercritical carbon dioxide (SC-CO₂) at 40°C with pressures of 5–30 MPa. At 40°C distribution coefficients of δ-lactones increased with pressure and exhibited molecular weight-dependency. Using experimental data, a mathematical model based on thermodynamic equilibria and material balance was developed and used to predict the distribution of inherent δ-lactones of AMF in a continuous SC-CO₂ extraction system. At 24.1 MPa/40°C column conditions, AMF was separated into a raffinate and 5 fractions. Predicted δ-lactones distribution coefficients in the raffinate and extract were in good agreement with experimental data.     

Continuous Supercritical Fluid Processing of Anhydrous Milk Fat in a Packed Column

A continuous supercritical carbon dioxide processing system was designed, built and operated to investigate its performance for fractionation of anhydrous milk fat (AMF). Packed columns showed mass transfer efficiencies five times greater than a spray column, Short- and medium-chain fatty acids were concentrated in the extract fractions and their amounts decreased as separation pressure increased. The ratio of unsaturated to saturated fatty acids in the raffinate fraction was 0.68 compared to 0.52 in the original AMF. The proportions of low-melting triglycerides in the range —42 to 15°C were greater in the extract fractions (44–72%) compared to the original AMF (29%).     

Separation/fractionation of triglycerides in terms of fatty acid constituents in palm kernel oil using supercritical CO2

Separation of triglycerides in terms of fatty acid constituents in palm kernel oil (PKO) from dehulled ground palm kernel using supercritical carbon dioxide (SC‐CO₂) as a solvent at 40 and 80 °C and pressures of 20.7, 27.6, 34.5, 41.4 and 48.3 MPa was studied. The extraction was continuous and PKO was fractionated into four fractions. Triglycerides with differing fatty acid contents of caprylic (C8), capric (C10), lauric (C12), myristic (C14), palmitic (C16), stearic (C18:0), oleic (C18:1) and linoleic (C18:2) acid were separated in different percentages in different fractions. About 70% of PKO comprises triglycerides with C8–C12 fatty acids and these fatty acids were mostly extracted in fractions 1and 2. As a result, higher yields were observed in these fractions, especially at pressures higher than 27.6 MPa. The C16 and C18:2 components were extracted in fractions 3 and 4. These triglycerides form only a part of the remaining 30% of the fatty acid composition of PKO, hence the evidently diminishing yield in the later fractions. However, the compositions were found to be statistically significant for higher pressures and fraction times according to the type of fatty acid constituents. Copyright © 2006 Society of Chemical Industry     

Adsorption and Desorption of Cholesterol in Continuous Supercritical Fluid Processing of Anhydrous Milk Fat

The adsorption breakthrough volume of adsorbent for cholesterol reduction of anhydrous milk fat (AMF) was determined by passing supercritical carbon dioxide (SC-CO₂) phase through an adsorption column with magnesium silicate at 40°C/241 bar. The breakthrough quantity for 76% cholesterol reduction was 2.0g fat/g adsorbent. With in-line adsorption on magnesium silicate, the cholesterol reduction in the extracts was 80 to 86%. Magnesium silicates saturated with cholesterol were regenerated with 10% ethanol in CO₂ at 40°C and 64°C/241 bar. The adsorption capacity of the magnesium silicate regenerated with 10% ethanol in SC-CO₂ at 40°C/241 bar was close to its original value. Its behavior was described by the Langmuir isotherm.     

Extraction of boldo (Peumus boldus M.) leaves with supercritical CO2 and hot pressurized water

High-activity fractions in boldo leaves were extracted with supercritical CO₂ (SC-CO₂) and hot pressurized water (HPW). Total yield after 3 h of extraction (0.6–3.5%) in low-pressure SC-CO₂ experiments increased with process pressure (60–150 bar) and decreased with temperature (30–60 °C), as expected. The extract obtained with SC-CO₂ at 50 °C and 90 bar contained approximately 50% of essential oil components. In higher pressure experiments with solvent mixtures, the yield increased with pressure (300–450 bar) and modifier concentration (2–10% ethanol), ranging 0.14–1.95 ppm for the alkaloid boldine and 1.8–4.8% for total solids following 1.5 h treatment at 50 °C. Boldine recovery was solubility-controlled, reaching 7.4 ppm after 7-h extraction at 450 bar and 50 °C using an ethanol–SC-CO₂ mixture (5% co-solvent). Boldine solubility and yield decreased when using pure CO₂ at higher pressure (600 bar, 50 °C). The extract yield after 3 h batch-wise HPW extraction increased from 36.9% at 100 °C to 53.2% at 125 °C, and then decreased as temperature was increased to 175 °C. Boldine yield decreased from 26.8 ppm at 100 °C to 0.7 ppm at 125 °C, and was negligible at ⩾150 °C. The essential oil yield increased to a maximum at 110 °C and was negligible at ⩾150 °C also. The ranking of antioxidant potency of various extracts was as follows: HPW at 110 °C > methanol (soxhlet extraction) ≫ high-pressure SC-CO₂ with or without polar co-solvent > low-pressure SC-CO₂.     

Effect of Olein Fractions of Milk Fat on Oxidative Stability of Ice Cream

The effect of low melting fractions of milk fat on oxidative stability of ice cream was investigated. Cream was fractionated at three different temperatures (25, 15, and 10°C), designated as LF-25, LF-15, and LF-10. All the low melting fractions were individually incorporated into ice cream and compared with a control, unmodified milk fat. The other ingredients were the same as in the control. Ice creams were stored at –18°C for six months and sampled every 30 days. Fractionation induced major changes in the fatty acid composition of all fractions. Concentration of short-chain and long chain unsaturated fatty acids increased in the low melting fractions. Peroxide value and anisidine value of LF-10 increased from 0.23 to 3.95 (meq/kg O₂) and 3.87 to 8.04. Conjugated dienes of control and LF-10 after six months were 1.39 and 4.72 at the same storage period. The flavor score of LF-10 was more than the control and remained indifferent from the control until two months of storage. After six months, the flavor score of LF-10 dropped by 3.6 points as compared to the control, 1.2 points. Low melting fractions of milk fat can be added in the formulation of ice cream to improve its nutritional value with acceptable sensory attributes. However, storage of ice cream formulated from low melting fractions is not recommended for over 60 days at –18°C.     

Extraction Conditions and Moisture Content of Canola Flakes as Related to Lipid Composition of Supercritical CO2 Extracts

Canola flakes obtained before and after conventional cooking were extracted with supercritical carbon dioxide (SC-CO₂) at 35–75°C and 20.7–62.0 MPa. Moisture content was adjusted to 12.7–42.5% (w/w, moisture-and oil-free basis). Moisture content of residual flakes after extraction was monitored. Amount of extract recovered increased with increasing temperature and pressure, except at 20.7 MPa pressure it decreased with temperature indicating a crossover of the solubility isotherms. Some water was co-extracted with canola oil. Free fatty acid (FFA) concentration of the extracts increased with decreasing pressure due to a decrease in triglyceride (TG) solubility. Oleic and linolenic acids had highest concentrations in the FFA fractions of all extracts. The lowest TG content of extracts was attained at 20.7 MPa and 75°C. The effect of extraction conditions was more pronounced than seed pre-treatments on yield and composition of SC-CO₂ extracts.     

Detection of milk fat adulteration with admixture of foreign oils and fats using a fractionation technique and the apparent solidification time test

A fractionation technique followed by the apparent solidification time (AST) test was adopted for detecting the admixture of foreign oils and fats in milk fat. The AST values of the solid fraction obtained at 20°C, and solid and liquid fractions obtained at 18°C for pure cow milk fat, were 2 min 30 s, and 3 min 21 s and 3 min 31 s, while for buffalo milk fat they were 1 min 58 s, and 2 min 47 s and 3 min 10 s respectively. This new approach can detect some mixtures of foreign oils and fats in cow milk fat but not in buffalo milk fat.     

Supercritical CO2 Extraction of Oil from Atlantic Mackerel (Scomber scombrus) and Protein Functionality

Supercritical carbon dioxide (SC-CO₂) extraction temperature and pressure for oil removal from freeze-dried, fall Atlantic mackerel muscle were optimized. The effect of extraction conditions on pH and water-binding potential (WBP) of the protein residue was evaluated. For the temperature range (35–55°C) and pressure interval (20.7–34.5 MPa), 34.5 MPa/35°C gave highest oil yield and concentration of ω-3 fatty acids. SC-CO₂ extraction at higher pressure levels caused slight decreases in protein residue pH. WBP of residual proteins was maximal at 1.49g H₂O/g protein for 34.5 MPa/45°C. The high lipid removal and retention of protein functionality in the residue after oil extraction may make such processes useful for future food applications.     

Modification of Crude Canola Lecithin for Food Use

Crude canola lecithin was fractionated by removal of neutral lipids with acetone followed by ethanol extraction with or without supercritical carbon dioxide (SC-CO₂). Two extraction steus of 60 min each at 35°C with 500 mL acetone were optimum for maximum yield of acetone solubles and maximum phospholipid (PL) concentration (66.7%) in the acetone insoluble (AI) fraction. AI were separated into 26.5% ethanol soluble and 68.7% ethanol insoluble fractions. PL concentration in the extract increased with increasing amounts of ethanol used. Yields were low for SC-CO₂/ethanol extraction, of which extracts from 55.2 MPa/ 70°C contained 32% PL (83% phosphatidyl choline).     

Temperature and pressure effects on supercritical carbon dioxide extraction of n-3 fatty acids from red seaweed

Lipids were extracted with supercritical carbon dioxide (SC-CO₂) from a subtropical red seaweed (Hypnea charoides) within the temperature range 40–50°C and the pressure range 24.1–37.9 MPa. In general, the extraction rates of algal lipids increased with pressure and temperature except when the pressure was at 24.1 MPa. The combined effect of pressure and temperature on the solubility of individual n-3 fatty acids in the SC-CO₂ varied with its carbon chain length. The concentrations of C₁₈, C₂₀ and C₂₂ n-3 fatty acids, extracted under different pressure and temperature conditions, were significantly different (p<0.05). Proportions of total polyunsaturated fatty acids increased significantly (p<0.05) and proportions of total saturated fatty acids decreased significantly (p<0.05) with increasing pressure as shown by the saturated/unsaturated and saturated/polyunsaturated ratios.     

Enzymatic interesterification of palm and coconut stearin blends

Hard fractions of palm oil and coconut oil, blended in the ratios of 90:10, 85:15, 80:20 and 75:25, were interesterified for 8 h using Lipozyme TL IM. Major fatty acids in the blends were palmitic acid (41.7–48.4%) and oleic acid (26.2–30.8%). Medium‐chain fatty acids accounted for 4.5–13.1% of the blends. After interesterification (IE), slip melting point was found to decrease from 44.8–46.8 °C to 28.5–34.0 °C owing to reduction in solids content at all temperatures. At 37.5 °C, the blends containing 25% coconut stearins had 17.4–19% solids, which reduced to 0.4–1.5% on IE, and the slip melting point (28.6 and 28.8 °C) indicated their suitability as margarine base. The reduction in solid fat index of the interesterified fats is attributed to the decrease in high‐melting triacylglycerols in palm oil (GS₃ and GS₂U type) and increase in triolein (GU₃) content from 1 to 9.2%. Retention of tocopherols and β‐carotene during IE was 76 and 60.1%, respectively, in 75:25 palm stearin and coconut stearin blend.     

Enrichment of the conjugated linoleic acid content of bovine milk fat by dry fractionation

This study investigated the effect of fat fractionation on the conjugated linoleic acid (cis-9, trans-11-C_18 : 2) content of bovine milk fat. Anhydrous milk fat was fractionated into hard and soft fractions using controlled cooling and agitation. Fractionation of milk fat pre-melted at 60°C using a temperature programme of 33–10°C and a cooling rate of 0.58°C h⁻¹ yielded a soft fraction containing 63.2% more conjugated linoleic acid (2.22 g 100 g⁻¹ FAME), which was also enriched in polyunsaturated fatty acids and vaccenic acid (trans-11-C_18 : 1) compared with the parent fat. Agitation following fractionation was found to have a negative effect on the conjugated linoleic acid content of the soft fraction. Refractionation of the soft fraction did not increase the yield of conjugated linoleic acid. The conjugated linoleic acid and trans fatty acid content of 26 selected food products ranging in milk fat content from 0 to 100% is reported. Conjugated linoleic acid concentrations ranged from 0 to 16.2 mg g⁻¹ fat and were generally lower than the trans fatty acid content which ranged from 0 to 155.7 mg g⁻¹ fat. Spreads containing vegetable oils contained higher trans fatty acid and lower conjugated linoleic acid contents than milk fat-containing products. This study highlights that a milk fat fraction enriched in conjugated linoleic acid may be achieved by dry fractionation.     

Extraction of Triglycerides and Phospholipids from Canola with Supercritical Carbon Dioxide and Ethanol

Phospholipids used as emulsifiers were sparingly soluble in supercritical CO₂ (SC-CO₂), but could be recovered with the addition of ethanol as an entrainer. Triglycerides and lipids containing phospholipids were extracted with SC-CO₂ in two subsequent steps from both canola flakes and press cake containing 0 and 5% ethanol, respectively. 70°C, 62.0 MPa and addition of ethanol at an initial level of 5% gave maximum yield of lipids. TLC showed the presence of phospholipids in SC-CO₂-ethanol extracts. SC-CO₂ extracts of flakes did not contain long chain fatty acids (C20:0, C22:0, C22:1), but they were extracted when ethanol was added to flakes and when press cake was the raw material.     

Treatment of milk fat with sn-2 specific Pseudozyma antarctica lipase A for targeted hydrolysis of saturated medium and long-chain fatty acids

Milk fat was treated with the sn-2 specific PAN-2 lipase from Pseudozyma antarctica with the aim of achieving selective removal of saturated fatty acids with chain lengths between C12 and C16, which are abundant at the sn-2 position. Hydrolysis was also carried out using the non-specific CRU-NS from Candida rugosa and the 1,3-specific TLA-1,3 from Thermomyces lanuginosus. PAN-2 treatment decreased the proportion of the target fatty acids considerably (content of 28%, w/w) and increased the proportion of unsaturated ones (content of 52%, w/w) at the degree of hydrolysis of 48% (mol/mol) in relation to the unhydrolysed substrate (47%, w/w, C12:0-C14:0 and 32%, w/w, unsaturated). The fatty acid distributions of the TLA-1,3 and CRU-NS treated samples were similar to untreated milk fat. The proportion of crystalline fat was smaller and the firmness lower in the PAN-2 treated milk fat samples than in the untreated milk fat at 5 °C.     

Supercritical Fluid Extraction of Dry-Milled Corn Germ with Carbon Dioxide

Dry-milled corn germ was extracted with supercritical carbon dioxide (SC-CO₂) at 5,000-8,000 psi and 50°C. CO₂-extracted oil was lower in free fatty acids and refining loss, and was lighter in color when compared with a commercial expeller-milled crude oil. Total unsaponifiable and tocopherol contents were similar for both oil types. The defatted, highly friable flour has a shelf-stable moisture content of 2-3% and good flavor quality. The flour contains 20% protein with good amino acid balance, meeting FAO specifications for food protein supplements. High pressure SC-CO₂ extraction also denatures the proteins, including oxidative enzymes. Peroxidase activity is reduced tenfold in SC-CO₂-extracted flour when compared with hexane-extracted flours. Storage tests for 5 wk at 38°C and for 2 months at 25°C show that flavor quality of untoasted SC-CO₂-defatted germ flour is maintained even under these extreme conditions.     

Supercritical carbon dioxide extraction of lutein from yellow maize (<Emphasis Type="Italic">Zea mays</Emphasis>) kernels: process optimization based on lutein content,antioxidant activity,and ω-6/ω-3 fatty acid ratio

Yellow maize kernels were subjected to supercritical carbon dioxide (SC-CO₂) extraction to obtain a lutein-rich extract with potential nutraceutical properties. SC-CO₂ extraction parameters (pressure and temperature) were optimized by employing a full-factorial (3²) design of experiments and response surface methodology, based on yield of lutein, antioxidant activity, and ω-6/ω-3 fatty acid ratio of the extracts. A Chrastil equation was also developed for predicting the solubility of lutein in SC-CO₂ under different extraction conditions. The optimized extraction condition was obtained at 500 bar, 70 °C for 90 min, at which the extract was found to possess a unique combination of the highest lutein yield (275.00 ± 3.50 μg/g of dry weight), along with a well-balanced ω-6/ω-3 fatty acid ratio (3:1). Moreover, the total phenol content and antioxidant activity were also found to be the highest at this condition. This lutein-rich extract is a promising nutraceutical or dietary supplement in the food industry.     

Extraction of oil from tiger nut (Cyperus esculentus L.) with supercritical carbon dioxide (SC-CO2)

Supercritical carbon dioxide (SC-CO₂) was used to extract oil from tiger nuts and the physicochemical properties and the impact of extraction conditions [i.e., temperature (40 °C–80 °C), pressure (20–40 MPa) and time (60–360 min)] on the oil yield were studied. The response surface analysis results revealed that the oil yield was significantly (P < 0.05) influenced by the main effect of the extraction pressure, extraction time and their quadratic effects respectively. However, the interaction between the extraction temperature and time had no significant (P > 0.05) effect on the oil yield. The highest oil yield was 26.28 g/100 g sample after 210 min of extraction time at 30.25 MPa and 60 °C respectively. The fatty acid composition of oils obtained by SC-CO₂ and Soxhlet showed marked variation. Also, the fatty acid composition varied depending on the operating conditions. The viscosity of the oil decreased with the increase in temperature.     

Composition of goat's milk fat triglycerides analysed by silver ion adsorption-TLC and GC–MS

The triglyceride (TG) composition of goat's milk fat (from the milk of five herds collected monthly, from November to May) was studied using AgNO₃-TLC and GC–MS. Two of the four fractions obtained by TLC contained trisaturated TGs (SSS) and represented 55% of the total TGs; they were separated by the chain length of short-chain fatty acid (FA). The TGs of the remaining fractions were identified as mono- (SSM) and polyunsaturated, representing 29 and 16% of the total TGs, respectively. The distribution of two SSS fractions by carbon number (CN) was unimodal, with maxima at CN40 and CN36 respectively. The proportions of SSM and polyunsaturated TG were high between CN38–CN44 and CN46–CN54, respectively (mean values, 35 and 52%). One hundred and twenty-four peaks (some containing more that one molecular species of TGs) were detected in the chromatogram of total fat; the sum of the 30 peaks representing >1 mol% was 70% of the total TGs. One hundred and thirty-seven molecular species were identified in the total goat's milk fat: 50% SSS, 30% SSM and 20% polyunsaturated. The most important in quantitative terms were medium-chain TGs containing C8, C10 or C12, and C18 : 1 as unsaturated FA.