Separation of squalene from olive oil deodorizer distillate using short-path molecular distillation

Squalene was recovered from an olive oil deodorizer distillate (OODD) containing 40% of squalene by a two-step process. The first step was to esterify the free fatty acids (FFAs) to make them less volatile. The second step was to separate the squalene by molecular distillation. The best esterification conditions were found to be 190°C and 360 min, where FFA content of the reaction mixture was reduced from 49.3% to 7.9%, however, an inevitable squalene loss (30%) was also observed due to a discontinuous operation. The remaining squalene (28%) in the esterified mixture was then distilled using a molecular distillation unit at elevated temperatures (190–230°C) and pressures (0.05–5 mmHg). When the temperature and vacuum during distillation increased, FFA content in the distillate reduced while distillate yield and squalene purity increased. The highest distillate yield (27.7%) and squalene purity (98.1%) were obtained at the highest applied temperature (230°C) under the lowest absolute pressure (0.05 mmHg), where FFA content of distillate was measured as 1.8%. High percentage of squalene (95%–98%) could be distilled at 230°C between 0.05 and 0.5 mmHg absolute pressures. The overall squalene recovery after all treatments was calculated as 68%.     

The concentration of tocols from rice bran oil deodorizer distillate using solvent

Tocols (tocopherols + tocotrienols) have been concentrated efficiently from rice bran oil (RBO) deodorizer distillate using solvent at low temperature. The levels of total tocols, total tocopherols, and total tocotrienols in RBO deodorizer distillate (starting material) were 31.5, 14.9, and 16.6 mg/g, respectively. Nine different solvents were tested, and acetonitrile was selected as the optimal solvent for concentrating tocols from the RBO deodorizer distillate. There was a significant (p <0.05) increase in the tocol level of the liquid fractions with decreasing temperature, for incubation temperatures up to –20 °C. In addition, significant differences (p <0.05) were observed in the relative percentages of α‐tocopherol, γ‐tocopherol, α‐tocotrienol, and γ‐tocotrienol between the raw sample and liquid fractions obtained at different temperatures using acetonitrile as the solvent. The concentration of the tocols from the RBO deodorizer distillate was temperature dependent, and a maximum of 89.9 mg/g was attained in the liquid fraction at – 40 °C. The relative percentage of tocotrienol homologs in the liquid fraction obtained at – 40 °C was approximately 80%. With acetonitrile as the solvent, the optimal temperature for concentrating the tocols from RBO deodorizer distillate was –20 °C when yield was considered.     

Optimization of supercritical CO2 extraction of different anatomical parts of lovage (Levisticum officinale Koch.) using response surface methodology and evaluation of extracts composition

Supercritical carbon dioxide extraction (SFE-CO₂) parameters were optimized using response surface methodology and central composite design for lovage (Levisticum officinale Koch.) roots and leaves containing valuable phytoconstituents. Mathematical model predicted the highest yields of extracts from roots and leaves 2.26 and 2.29%, respectively, at 45 MPa pressure, 60 °C temperature, 90 min (roots) and 30 min (leaves) extraction time, whereas the yield of hydrodistilled essential oil was 0.24 and 0.74%, respectively. The highest relative content of the most valuable constituent Z-ligustilide in roots and leaves extracts was 77 and 50% at 10 MPa; however, the highest yields of this compound from 100 g of dry material were obtained at the highest applied pressure and constituted 1188 mg (roots) and 540 mg (leaves). This study showed that lovage is a good source of Z-ligustilide and SFE-CO₂ is a preferable technique for its isolation.     

Experimental optimization and mathematical modeling of supercritical carbon dioxide extraction of essential oil from Pogostemon cablin

The supercritical carbon dioxide extraction was applied to obtain essential oil from Pogostemon cablin in this work. Effect of extraction parameters including temperature, pressure, extraction time and particle size on extraction yield was investigated, and the response surface methodology with a Box-Behnken Design was used to achieve the optimized extraction conditions. The maximum yield of essential oil was 2.4356% under the conditions of extraction temperature 47℃, pressure 24.5 MPa and extraction time 119 min. Moreover, based on the Brunauer-Emmett-Teller theory of adsorption, a mathematical modeling was performed to correlate the measured data. The model shows a function relationship between extraction yield and time by a simple equation with three significantly adjustable parameters. These model parameters have been optimized through simulated annealing algorithm. The predicted data from the mathematical model show a good agreement with the experimental data of the different extraction parameters.     

Application of short path distillation for recovery of polyphenols from deodorizer distillate

During physical refining of oil derived from ‘high temperature short time’ (HTST) pretreated rapeseeds, polyphenols are separated from the oil during deodorization and accumulate together with other high‐value minor compounds in the so‐called deodorizer distillate. For recovery of these compounds single‐stage and multistage short path distillations were carried out in a laboratory scale apparatus at evaporation temperatures between 110 and 170°C and pressures between 0.006 and 0.01 mbar. In addition, the removal of traces of pesticides from rapeseed deodorizer distillate was investigated. It was observed that these compounds can be separated from deodorizer distillate by means of short path distillation very effectively. On the basis of these experiments, a recovery process for polyphenols was proposed involving short path distillation, acid catalyzed esterification with methanol, solvent crystallization and solvent extraction processes. The final product was a polyphenol enriched extract containing about 14% of polyphenols. A polyphenol recovery of 50% is considered to be reachable and fractions rich in tocopherols and sterols may be obtained as by‐products.     

A single-step isolation of squalene from olive oil deodorizer distillates by using centrifugal partition chromatography

High added-value squalene (SQ) was purified in one step from olive oil deodorizer distillates (OODD) using preparative centrifugal partition chromatography (CPC) method, operating in the dual mode. The fractionation was performed using a non-aqueous biphasic solvent system consisting of heptane–acetonitrile–butanol (1.8:1.4:0.7, v/v/v), leading to the isolation of the target compound in 4 hours, using a preparative 1L column. Furthermore, a fast UHPLC-DAD method was developed and applied for the identification and quantification of SQ in both OODD and purified form. The content of SQ in the initial material was 23.4%, while the purity of the isolated SQ was 95.5%. The recovery of SQ was calculated at 76.3%. The productivity of the process was calculated at 234 mg/h/L.     

Squalene recovery from olive oil deodorizer distillates

Olive oil deodorization distillate contains squalene in a concentration range of 10 to 30 wt%. A process for its recovery by supercritical carbon dioxide extraction is described. The process consists mainly of converting the free fatty acids and the methyl and ethyl esters normally occurring in this by-product into their corresponding triglycerides. The latter are then extracted with supercritical carbon dioxide to provide a highly enriched squalene fraction. The process has been carried out on a pilot-plant scale with a column operating in the contercurrent mode. The relationship between the experimental conditions and squalene purity and yield has been studied. Analytical methods were used for the determination of squalene and other components in the fractions. By use of this process, squalene can be recovered in high purity and yields of about 90%.     

Continuous production of fatty acid methyl esters from corn oil in a supercritical carbon dioxide bioreactor

Continuous production of fatty acid methyl esters (FAMEs) from corn oil was studied in a supercritical carbon dioxide (SC-CO₂) bioreactor using immobilized lipase (Novozym 435) as catalyst. Response surface methodology (RSM) based on central composite rotatable design (CCRD) was employed to investigate and optimize the reaction conditions: pressure (11-35 MPa), temperature (35-63 °C), substrate mole ratio (methanol:corn oil 1-9) and CO₂ flow rate (0.4-3.6 L/min, measured at ambient conditions). Increasing the substrate mole ratio increased the FAME content, whereas increasing pressure decreased the FAME content. Higher conversions were obtained at higher and lower temperatures and CO₂ flow rates compared to moderate temperatures and CO₂ flow rates. The optimal reaction conditions generated from the predictive model for the maximum FAME content were 19.4 MPa, 62.9 °C, 7.03 substrate mole ratio and 0.72 L/min CO₂ flow rate. The optimum predicted FAME content was 98.9% compared to an actual value of 93.3 ± 1.1% (w/w). The SC-CO₂ bioreactor packed with immobilized lipase shows great potential for biodiesel production.     

Enzymatic methylation of canola oil deodorizer distillate

Methylation of canola oil deodorizer distillate catalyzed by a nonspecific lipase was investigated. The conversion of fatty acids to methyl esters has been optimized by using a statistical design. Up to 96.5% conversion of fatty acids to their methyl esters has been achieved without the aid of vacuum or any water-removing agent. The effects of temperature, ratio of the reactants (methanol: fatty acids in the deodorizer distillate) and enzyme concentration on the equilibrium conversion were studied. The temperature and ratio of the reactants showed a significant effect on the conversion of fatty acids to methyl esters and they exhibited a strong interactive effect. Enzyme concentration in the range of 2.7% to 4.3% did not show a significant effect on the equilibrium conversion of fatty acids. Greater than 95% conversion of fatty acids to methyl esters was achieved at temperatures around 50°C and at a ratio of the reactants between 1.8 and 2.0. The inhibitory effect of hydrophilic methanol on the enzyme activity was largely reduced by working at the lower temperature range (around 50°C).     

Enzymatic pretreatment of deodorizer distillate for concentration of sterols and tocopherols

Separation of sterols and tocopherols from fatty acids in deodorizer distillate was facilitated through lipase-catalyzed modification of fatty acids in canola, mixed and soya deodorizer distillates. The fatty acid esterification with methanol catalyzed by SP-382 (an immobilized nonspecific lipase) proceeded rapidly, with conversion of fatty acid to methyl ester in 5 h being 96.5, 83.5 and 89.4%, respectively. A model mixture of pure oleic acid and dl-α-tocopherol was used to study any potential side reactions that may lower the tocopherol content during the esterification reaction. Under the conditions employed, the loss of tocopherol was less than 5%. Simple vacuum distillation (1–2 mm Hg) was employed to remove the volatile fraction (methyl esters of fatty acids, some fatty acids and other volatiles) of the esterified deodorizer distillate, leaving behind sterols, sterol esters and tocopherols. Sterols and tocopherols were almost completely retained in the residue fraction with recoveries in the range of 95%. Overall recoveries of sterols and tocopherols after esterification and distillation were over 90% for all the deodorizer distillate samples.     

Separation of squalene from palm fatty acid distillate using adsorption chromatography

A method to separate squalene from palm fatty acid distillate (PFAD) using neutralization‐hydrolysis‐neutralization before employing adsorption column chromatography was developed. Extraneous matters, especially free fatty acids (83.8%) and acylglycerols (12.7%), were first neutralized and removed before being subjected to hydrolysis by using commercially available, immobilized Candida antartica lipase, at 65 °C for 8 h. Neutralization followed by hydrolysis and repeated neutralization successfully concentrated squalene from an initial amount of 3.76% to 27.5%. Oil extracted from neutralized‐hydrolyzed‐neutralized PFAD was then passed through a Diaion HP‐20 column using reverse‐phase adsorption chromatography. Squalene was desorbed by hexane, with a recovery of 93%.     

Supercritical carbon dioxide extraction of squalene and tocopherols from amaranth and assessment of extracts antioxidant activity

Squalene and tocopherols are the most important bioactive constituents in lipophilic amaranth fraction. Therefore, developments of processes of isolation of amaranth extracts enriched with these compounds are of interest. In this study the lipophilic fraction of amaranth seeds was extracted by supercritical fluid extraction with carbon dioxide (SCE-CO₂) under different pressure conditions and by adding 2 and 5% of cosolvent ethanol. The yield of extract varied from 1.37 (15 MPa without cosolvent) to 5.12% (55 MPa and 5% of cosolvent). The highest content of unsaponifiables (21.1%) in the extract was at 55 MPa and 5% of cosolvent; at these conditions the yields of tocopherols and squalene from amaranth seeds were 317.3 mg/kg and 0.289 g/100 g, respectively. Tocopherol isomers in amaranth oil were distributed at the approximate ratio of 1(α-T):27(β-T):6.5(γ-T):5(δ-T). The extract was fractionated in the two separators by gradual decrease of the pressure and it was found that the fraction obtained at ambient conditions contained the highest concentration of tocopherols (up to 7.6 mg/g) and squalene (up to 17.9 g/100 g oil). The highest antioxidant activity measured by the L-ORAC assay possessed the fractions with the highest concentrations of squalene and tocopherols and obtained at 15 MPa with pure CO₂ (235.1 μmol TE/g) and 2% of cosolvent (257.6 μmol TE/g).     

Facile purification of tocopherols from soybean oil deodorizer distillate in high yield using lipase

Tocopherols have been purified from deodorizer distillate produced in the final deodorization step of vegetable oil refining by a process including molecular distillation. Deodorizer distillate contains mainly tocopherols, sterols, and free fatty acids (FFA); the presence of sterols hinders tocopherol purification in good yield. We found that Candida rugosa lipase recognized sterols as substrates but not tocopherols, and that esterification of sterols with FFA could be effected with negligible influence of water content. Enzymatic esterification of sterols with FFA was thus used as a step in tocopherol purification. High boiling point substances including steryl esters were removed from soybean oil deodorizer distillate by distillation, and the resulting distillate (soybean oil deodorizer distillate tocopherol concentrate; SODDTC) was used as a starting material for tocopherol purification. Several factors affecting esterification of sterols were investigated, and the reaction conditions were determined as follows: A mixture of SODDTC and water (4∶1, w/w) was stirred at 35°C for 24 h with 200 U of Candida lipase per 1 g of the reaction mixture. Under these conditions, approximately 80% of sterols was esterified, but tocopherols were not esterified. After the reaction, tocopherols and FFA were recovered as a distillate by molecular distillation of the oil layer. To enhance further removal of the remaining sterols, the lipase-catalyzed reaction was repeated on the distillate under the same reaction conditions. As a result, more than 95% of the sterols was esterified in total. The resulting reaction mixture was fractionated to four distillates and one residue. The main distillate fraction contained 65 wt% tocopherols with low contents of FFA and sterols. In addition, the residue fraction contained high-purity steryl esters. Because the process presented in this study includes only organic solvent-free enzymatic reaction and molecular distillation, it is feasible as a new industrial purification method of tocopherols. This work was presented at the Biocatalysis symposium in April 2000, held at the 91st Annual Meeting and Expo of the American Oil Chemists Society, San Diego, CA.     

Production of phytosterol esters from soybean oil deodorizer distillates

Enzymatic esterification and supercritical fluid extraction was used to produce phytosterol esters from soybean oil deodorizer distillates. The raw material was first subjected to a two‐step enzymatic reaction; the product obtained mainly comprised fatty acid ethyl esters, tocopherols and phytosterol esters, together with minor amounts of squalene, free fatty acids, free sterols and triacylglycerols. The phytosterol esters were then purified from this mixture using supercritical carbon dioxide. Experimental extractions were carried out in an isothermal countercurrent column (without reflux), with pressures ranging from 200 to 280 bar, temperatures of 45–55 °C and solvent‐to‐feed ratios from 15 to 35 kg/kg. Using these extraction conditions, the fatty acid esters were completely extracted and, thus, the fractionation of tocopherols and phytosterol esters was studied. At 250 bar, 55 °C and a solvent‐to‐feed ratio of 35, the phytosterol esters were concentrated in the raffinate up to 82.4 wt‐% with satisfactory yield (72%).     

菜籽油脱臭馏出物中生物柴油的排放特性研究

通过分子蒸馏技术从菜籽油脱臭馏出物中制备生物柴油,并通过对比试验,研究了柴油机燃烧生物柴油和普通柴油对发动机经济性和排放特性的影响。研究结果表明,脱臭馏出物中制备的生物柴油与普通的0#柴油性质相似,脂肪酸甲酯质量分数在90%以上。在生物柴油的排放中,除CO2排放和耗油率相对升高,CO2排放增加幅度达到20%左右;CO,CH和碳烟都相对降低,烟度和CH最高降幅分别达到54%和88%;NOx排放在高载荷阶段体积分数上升。     

利用菜籽油脱臭馏出物制备生物柴油新工艺

以菜籽油脱臭馏出物为原料,首先以D002阳离子交换树脂作催化剂,进行酯化反应,降低原料酸值;然后以氢氧化钾作催化剂进行醇解反应来制备生物柴油的二步法新工艺路线。结果表明:D002阳离子交换树脂具有很强的催化活性,游离脂肪酸最高转化率达97.7%,连续使用4次后,催化活性仍然很高,达96%以上;碱催化过程中甘油酯的最高转化率达97.4%。产品品质大都符合美国ASTMD6751-03生物柴油标准。由此可见,先用树脂催化处理高酸值废油,然后进行碱催化制备生物柴油二步法工艺是一种切实可行的方法。     

Preparation of high purity squalene from soybean oil deodorizer distillate with the combination of macroporous resin and thin-film evaporation coupling distillation

ABSTRACT

In this study, the combination of macroporous adsorbent resins (MARs) and thin-film evaporation coupling distillation (TFECD) was studied systematically, with aim to obtain high value-added squalene from soybean oil deodorizer distillate (SODD). Five types of resins (X-5, D-101, D4020, DM301, and AB-8) were first used to evaluate the adsorption/desorption properties of squalene. D101 resin exhibited higher adsorption/desorption capacities and desorption ratios for squalene based on static adsorption results among the tested resins. We further investigated the adsorption kinetics, isotherms, and thermodynamics with D101 resin as adsorbent. The adsorption of squalene on D101 was best fitted to pseudo-second-order kinetic model and the Langmuir equation. The dynamic adsorption and desorption indicated that similar results were observed in the static adsorption test. The purity of squalene was increased from 7.5 to 82.5% with the recovery up to 88.5% after separation on D101 column. The resin-refined sample was directly subjected to TFECD purification. Under optimized process parameters, the final product with purity of 98.5% and recovery yield of 76.5% was confirmed by high-performance liquid chromatography (HPLC) analysis. The present study provided an effective method for large-scale production of high purity squalene.     

Purification of steryl esters from soybean oil deodorizer distillate

Soybean oil deodorizer distillate (SODD) contains steryl esters in addition to tocopherols and sterols. Tocopherols and sterols have been industrially purified from SODD but no purification process for steryl esters has been developed. SODD was efficiently separated to low b.p. substances (including tocopherols and sterols) and high b.p. substances (including 11.2 wt% DAG, 32.1 wt% TAG, and 45.4 wt% steryl esters) by molecular distillation. The high b.p. fraction is referred to as soybean oil deodorizer distillate steryl ester concentrate (SODDSEC). We attempted to purify steryl esters after a lipase-catalyzed hydrolysis of acylglycerols in SODDSEC. Screening of industrially available lipases indicated that Candida rugosa lipase was most effective. Based on the study of several factors affecting hydrolysis, the reaction conditions were determined as follows: ratio of SODDSEC/water, 1∶1 (w/w); lipase amount, 15 U/g reaction mixture; temperature, 30°C. When SODDSEC was agitated for 24 h under these conditions, acylglycerols were almost completely hydrolyzed and the content of steryl esters did not change. However, study with a mixture of steryl oleate/trilinolein (1∶1, w/w) indicated that about 20% of constituent FA in steryl esters were exchanged with constituent FA in acylglycerols. Steryl esters in the oil layer obtained by the SODDSEC treatment with lipase were successfully purified by molecular distillation (purity, 97.3%; recovery, 87.7%).