Fast Production of Diacylglycerol in a Solvent Free System via Lipase Catalyzed Esterification Using a Bubble Column Reactor

In this study, diacylglycerols (DAG) were synthesized rapidly (~30 min) in a solvent‐free system via esterification of glycerol with fatty acids (FA, the mixture of 60 wt% palm oil deodorizer distillate and 40 wt% oleic acid) catalyzed by Lipozyme 435 (Novozymes A/S, Copenhagen, Denmark) using a bubble column reactor. The content of DAG, monoacylglycerols (MAG), triacylglycerols (TAG) and free fatty acids (FFA) in the crude product were 57.94 ± 1.60 wt%, 24.68 ± 2.08 wt%, 2.67 ± 1.72 wt% and 14.69 ± 1.22 wt%, respectively under the selected conditions, which were enzyme load of 5.0 wt%, glycerol/FA mole ratio of 7.5, initial water content of 2.5 wt%, reaction temperature of 60 °C, reaction time of 30 min and N₂ gas flow of 10.6 cm min^?1. The final product containing 91.30 ± 1.10 wt% of DAG was obtained by one‐step molecular distillation at 200 °C. The reusability of Lipozyme 435 was investigated by evaluating the esterification degree (ED) and the DAG content in the crude products in 30 successive runs. The enzyme retained 95.10 % of its original activity during 30 successive runs according to comparison of the ED. The new process showed a very high efficiency in production of DAG with a high purity. The ratio of positional isomers 1,3‐DAG to 1,2 ‐DAG was 2:1 in the final product. The certain plasticity (melting point of 44 °C) and content of unsaturated fatty acids made the product a valuable food ingredient.     

Crystallization Behavior of Diacylglycerol-Rich Oils Produced from Rapeseed Oil

We examined the crystallization behavior of high-melting fractions in liquid oil containing high concentrations of diacylglycerols (DAG >80%) (hereafter referred to as DAG-rich oil). By differential scanning calorimetry and optical microscopy at moderate cooling rates, crystallization in the DAG-rich oil was detected at around 6 °C. It was found that the crystallization extent increased with decreasing temperatures of crystallization below 0 °C. A gas chromatographic analysis was performed on the crystallized fractions, which were separated by filtration at different periods of isothermal crystallization at 3 °C. The results indicated that at earlier crystallization periods, the concentrations of 1,3-disaturated DAG such as palmitic and stearic acid moieties (15 min) and 1,3-saturated–unsaturated mixed-acid DAG including oleic acid, palmitic, and stearic acid moieties (15 min–3 h) were predominant. However, the concentrations of 1,3-diunsaturated DAG including oleic acid moiety increased after a crystallization period of 6 h. To clarify the sequential crystallization process of DAG, we examined the binary mixing behavior of principal DAG components occurring in the rapeseed-based DAG-rich oil. It was evident that 1,3-disaturated DAG, 1,3-saturated–unsaturated mixed-acid DAG, and 1,3-diunsaturated DAG exhibited immiscible behavior. From these data, basic information on the precipitation processes in DAG-rich oils at chilled temperatures was obtained.     

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%.     

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.     

Melting behavior of blends of milk fat with hydrogenated coconut and cottonseed oils

The melting behavior of milk fat, hydrogenated coconut and cottonseed oils, and blends of these oils was examined by nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC). Solid fat profiles showed that the solid fat contents (SFC) of all blends were close to the weighted averages of the oil components at temperatures below 15°C. However, from 15 to 25°C, blends of milk fat with hydrogenated coconut oils exhibited SFC lower than those of the weighted averages of the oil components by up to 10% less solid fat. Also from 25 to 35°C, in blends of milk fat with hydrogenated cottonseed oils, the SFC were lower than the weighted averages of the original fats. DSC measurements gave higher SFC values than those by NMR. DSC analysis showed that the temperatures of crystallization peaks were lower than those of melting peaks for milk fat, hydrogenated coconut oil, and their blends, indicating that there was considerable hysteresis between the melting and cooling curves. The absence of strong eutectic effects in these blends suggested that blends of milk fat with these hydrogenated vegetable oils had compatible polymorphs in their solid phases. This allowed prediction of melting behavior of milk-fat blends with the above oils by simple arithmetic when the SFC of the individual oils and their interaction effects were considered.     

Diacylglycerols from butterfat: Production by glycerolysis and short-path distillation and analysis of physical properties

The aim of this paper was to develop a process for the production of DAG from butterfat through glycerolysis and short-path distillation and to evaluate the physical properties of the DAG in comparison with the original butterfat. Chemical glycerolysis produced a mixture of acylglycerols containing DAG together with MAG and TAG. From the mixture of glycerolysis products, MAG were removed through three consecutive distillations (vacuum <0.001 mbar) at 150°C. TAG were separated from DAG by distillation at 210°C, which gave a product with more than 80% DAG in the distillates. Distillation temperatures had significant effects on acyl migration. The formation of desirable 1,3-DAG was favored at higher temperatures. Under 210°C distillation, the equilibrium ratio of 6∶4 was obtained between 1,3-DAG and 1,2(2,3)-DAG. The FA profile of the DAG product was relatively similar to the original butterfat. The total DAG recovery was around 77% in the pilot-scale production. The different patterns of m.p. were observed between butterfat and the DAG fraction produced as well as the MAG fraction collected. Solid fat content profiles of the DAG fraction and its mixtures with rapeseed oil possessed trends similar to those of the corresponding butterfat and its mixtures with rapeseed oil. Compared with butterfat, the DAG fraction behaved differently in its thermal profiles, crystallization patterns, and rheological properties; for example, the dropping point was 13°C higher for the latter than for the former, and the crystal pattern was mostly β form for the latter, whereas the former was the β′ form.     

Crystallization Behavior of High Behenic Acid Stabilizers in Liquid Oil

The crystallization behavior and structure of mixtures of a high behenic acid stabilizer (HBS) in peanut oil, high oleic safflower oil and sesame oil were studied in order to elucidate the mechanism behind liquid oil stabilization. Both the chemical composition of the oil and cooling rate influenced the crystallization behavior and structure of HBS. The critical gelation concentration of HBS ranged from 6.5% for peanut oil crystallized at 3 °C/min to 11% for sesame oil mixtures crystallized at 0.6 °C/min. The free energy of nucleation (ΔG) was the highest for sesame oil (142 kJ/mol) followed by high oleic safflower oil (75.8 kJ/mol) and peanut oil (15.9 kJ/mol). The HBS peanut oil mixture displayed the highest storage modulus (G′) under both cooling rates studied. In general, HBS-oil mixtures crystallized at a higher cooling rate exhibited high SFC values, lower crystallization temperatures and a predominance of the β′ polymorph, and they had a microstructure characterized by uniformly sized spherulites. In contrast, slow cooling rates led to higher critical gelation concentrations of HBS, lower SFC, fractionation of higher melting and lower melting fractions, a more stable polymorphic form β and a wide range of spherulite sizes.     

Unique Homo-Crystallization and Melting Behavior of Poly(L-lactic acid): Influence of Stereocomplex with Varied Structure

Homo-crystallization and melting behavior of poly(L-lactic acid) (PLLA) with poly(D-lactic acid) (PDLA) (≤10 wt.%) was studied. The different thermal history had been applied to exert structural variation on stereocomplex (SC). The PLLA/PDLA blend showed different crystallization and melting behavior when cooled from 250°C or 200°C. Double melting peaks were observed after the blend was cooled from 250°C. SC annealing at different temperatures exhibited significant effect for melt-crystallization of PLLA. Influence of initial melting condition before cooling was also investigated. The cold crystallization of amorphous blend initially was studied and some novel results had been observed.     

Influence of Enzymatic Remediation on Compositional and Thermal Properties of Palm Oil and Palm Oleins from Dry Fractionation

Characteristics of crude palm oil are high FFA and DAG contents. High DAG content may affect throughput and yield during fractionation: high‐grade specialty fats such as hard palm mid fraction require premium crude palm oil to secure adequate crystallization properties. Moreover, DAGs are generally considered the main precursors for the formation of glycidyl esters during high‐temperature deodorization. The purpose of this study was to investigate the effect of enzymatic remediation on the reduction of FFA and DAG in crude palm oil. In practice, series of process parameters (vacuum and reaction time) were investigated, and the quality of enzymatically remediated crude palm oils was examined in terms of FFA and DAG reduction, TAG composition, and SFC and DSC melting profiles. Fully refined enzymatically remediated palm oils were then dry fractionated. The quality of the oleins derived from the enzymatically remediated palm oil was compared to that of regular RBD palm oleins.     

Production of FAME from acid oil,a by-product of vegetable oil refining

Simple alkyl FA esters have numerous uses, including serving as biodiesel, a fuel for compression ignition (diesel) engines. The use of acid-catalyzed esterification for the synthesis of FAME from acid oil, a by-product of edible vegetable oil refining that is produced from soapstock, was investigated. Soybean acid oil contained 59.3 wt% FFA, 28.0 wt% TAG, 4.4 wt% DAG, and less than 1% MAG. Maximum esterification occurred at 65°C and 26 h reaction at a molar ratio of total FA/methanol/sulfuric acid of 1∶15∶1.5. Residual unreacted species under these conditions, as a fraction of their content in unesterified acid oil, were FFA, 6.6%; TAG, 5.8%; and DAG, 2.6%. This corresponds to estimated concentrations of FFA, 3.2%; TAG, 1.3%; and DAG, 0.2%, on a mass basis, in the ester product. In an alternative approach, the acylglycerol species in soapstock were saponified prior to acidulation. High-acid (HA) acid oil made from this saponified soapstock had an FFA content of 96.2 wt% and no detectable TAG, DAG, or MAG. Optimal esterification conditions for HA acid oil at 65°C were a mole ratio of FFA/methanol/acid of 1∶1.8∶0.17, and 14 h incubation. FAME recovery under these conditions was 89% of theoretical, and the residual unesterified FFA content was approximately 20 mg/g. This was reduced to 3.5 mg/g, below the maximum FFA level allowed for biodiesel, by washing with NaCl, NaHCO₃, and Ca(OH)₂ solutions. Alternatively, by subjecting the unwashed ester layer to a second esterification, the FFA level was reduced to less than 2 mg/g. The acid value of this material exceeded the maximum allowed for biodiesel, but was reduced to an acceptable value by a brief wash with 0.5 N NaOH.     

Isolation of tocopherol and sterol concentrate from sunflower oil deodorizer distillate

The isolation of tocopherols and sterols together as a concentrate from sunflower oil deodorizer distillate was investigated. The sunflower oil deodorizer distillate was composed of 24.9% unsaponifiable matter with 4.8% tocopherols and 9.7% sterols, 28.8% free fatty acid (FFA) and 46.3% neutral glycerides. The isolation technology included process steps such as biohydrolysis, bioesterification and fractional distillation. The neutral glycerides of the deodorizer distillates were hydrolyzed byCandida cylindracea lipase. The total fatty acids (initial FFA plus FFA from neutral glycerides) were converted into butyl esters withMucor miehei lipase. The esterified product was then fractionally distilled in a Claisen-vigreux flask. The first fraction, which was collected at 180–230°C at 1.00 mm of Hg for 45 min, contained mainly butyl esters, hydrocarbons, oxidized products and some amount of free fatty acids. The fraction collected at 230–260°C at 1.00 mm Hg for 15 min was rich in tocopherols (about 30%) and sterols (about 36%). The overall recovery of tocopherols and sterols after hydrolysis, esterification and distillation were around 70% and 42%, respectively, of the original content in sunflower oil deodorizer distillate.     

Hydrofining of Oil Shale Pyrolysis Tar in the Presence of Ultradispersed Catalysts

The results of the catalytic hydroconversion of oil shale pyrolysis tar in the presence of in situ and ex situ synthesized ultradispersed and nanosized MoS₂ particles in a flow reactor at a hydrogen pressure of 7MPa, a temperature of 430–450°C, a feed space velocity of 1–2.9 h^–1, and an H₂/feed flow ratio of 1000 L(NTP)/L with and without recycling the unconverted residue are reported. The optimum conditions of hydroconversion under which the yield of distillate fractions was 91.9 wt % on a raw material basis are proposed. It is shown that the quality of the refined synthetic oil makes it possible to use it in traditional petrochemical processes.     

Palmitoleic Acid Enrichment of Seabuckthorn (Hippophaë rhamnoides L.) Pulp Oil by Crystallization Process

Abstract

Seabuckthorn pulp oil was fractionated using a crystallization process with acetone under controlled cooling rate of 0.25°C/min without agitation at different crystallization temperatures ranging from ? 15 to 15°C. The obtained liquid (LF) and solid (SF) fractions were analyzed for their fatty acid and triacylglycerol compositions and their melting profiles were characterized. Fractionation at ? 15°C yielded about 20% of LF where palmitoleic acid represented 53% of total lipids. The SF fraction was mainly rich in palmitic acid. LF were richer in triacylglycerol with acyl carbon numbers of 50 and 52 (C50 and C52) than SF, which contained a higher amount of C48. The melting curves of LF and SF showed multiple endothermic transitions.     

Fractionation of squid visceral oil ethyl esters by short-path distillation

Squid visceral oil contains high levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Its ethyl esters were fractionated by short-path distillation in this study. The elimination temperatures of squid visceral oil ethyl esters (SVOEE) ranged from 50 to 140°C, increasing with the carbon number of ethyl esters. The elimination temperature of cholesterol was higher than those of SVOEE. The SVOEE of Illex argentinus (SVOEE-A) was more advantageous as the raw material (feed) than that of Ommastrephes bartrami (SVOEE-B) for the isolation of EPA and DHA, because SVOEE-A contained less 20∶1 and 22∶1. When SVOEE-A originally containing 9.0% EPA, 14.7% DHA, and 1,121 mg/100 g of cholesterol was distilled from 50 to 150°C with 20°C interval, the 130°C distillate could give 15.5% EPA and 34.7% DHA with 99 mg/100 g of cholesterol, and the yield was 21.8%. The 150°C distillate could give 43.1% DHA with 496 mg/100 g of cholesterol. Furthermore, the distillates collected from 110 to 150°C contained 24.4 to 50.2% of EPA plus DHA, and their total yield was 58.3%. The final residue after 150°C distillation contained 77% of the total cholesterol in the initial SVOEE-A, and the yield was 6.0%.     

Response surface methodology applied to optimization of distilled monoglycerides production

This work demonstrates that response surface methodology (RSM) is a powerful tool for the optimization of the production of distilled MG. Experiments with a centrifugal molecular distillator having an evaporation area of 0.0046 m² were carried out using RMS to identify operating conditions that can lead to higher MG purity. The independent variables studied were the evaporator temperature (TEV) and the volumetric feed flow rate (Q). The experimental range was from 100 to 300°C for TEV and between 5 and 15 mL/min for Q. High-performance size exclusion chromatography was used to evaluate TG, DG, MG, FFA, and glycerol (GL) compositions. Results were presented as MG concentration surfaces. Starting from a material with 10.8% of TG, 37.7% of DG, 43.6% of MG, and 7.2% of GL, the maximum MG, purity in the distillate stream with just one distillation step was 82.6% at a TEV equal to 250°C and Q equal to 5 mL/min. At these conditions, the MG recovery was 61%. A strategy was developed to obtain distilled MG with 96.3% purity.     

Comparison of lipase-transesterified blend with some commercial solid frying shortenings in Malaysia

A transesterified experimental solid frying shortening was prepared from a palm stearin/palm kernel olein blend at 1∶1 ratio (by weight) by using Rhizomucor miehei lipase at 60°C for 6 h. The fatty acid (FA) and triacylglycerol compositions, polymorphic forms, melting and cooling characteristics, slip melting point (SMP), and solid fat content (SFC) of the transesterified blend were then compared with five commercial solid frying shortenings (three domestic and two imported) found in Malaysia. All the domestic shortenings contained nonhydrogenated palm oil or palm olein and palm stearin as the hard stock, whereas the imported frying shortenings were formulated from soybean oil and cottonseed oil and contained high level of β′ crystals. Trans FA were also found in these samples. The lipase-transesterified blend was found to be more β′-tending than the domestic samples. The SMP of the transesterified blend (47.0°C) fell within the range of the domestic samples (37.8–49.7°C) but was higher than the imported ones (42.3–43.0°C). All samples exhibited similar differential scanning calorimetry cooling profiles, with a narrow peak at the higher temperatures and a broad peak at the lower temperatures, even though their heating thermograms were quite different. Imported samples had flatter SFC curves than both the experimental and domestic samples. The domestic samples were found to have better workability or plasticity at higher temperatures than the imported ones, probably because they were formulated for a tropical climate.     

Refining high-free fatty acid wheat germ oil

Wheat germ oil was refined using conventional degumming, neutralization, bleaching, and continuous tray deodorization, and the effects of processing conditions on oil quality were determined. The crude wheat germ oil contained 1,428 ppm phosphorus, 15.7% free fatty acid (FFA), and 2,682 ppm total tocopherol, and had a peroxide value (PV) of 20 meq/kg. Degumming did not appreciably reduce the phosphorus content, whereas neutralization was effective in removing phospholipid. Total tocopherol content did not significantly change during degumming, neutralization, and bleaching. A factorial experimental design of three deodorization tempeatures and three residence times (oil flow rates) was used to determine quality changes during deodorization. High temperatures and long residence times in deodorization produced oils with less FFA, PV, and red color. Deodorization at temperatures up to 250°C for up to 9 min did not significantly reduce tocopherol content, but, at 290°C for 30-min residence time, the tocopherol content was significantly reduced. Good-quality wheat germ oil was produced after modifying standard oil refining procedures.     

Purification and deodorization of structured lipids by short path distillation

Purification of structured lipids (SL), produced from lipase‐catalyzed acidolysis of rapeseed oil and capric acid, and deodorization of randomized SL, produced from chemical randomization of fish oil and tricaprin, were studied in a bench‐scale short path distillation (SPD). SL obtained from enzymatic acidolysis usually contain a large proportion of medium‐chain and long‐chain free fatty acids. Two SPD steps have been applied for the removal of free fatty acids. Parameters such as evaporator temperature, feeding flow rate, stirring roller speed, and the content of free fatty acids (FFA) added to the starting materials were optimized with respect to FFA left in the product residuals and to tocopherol loss from the starting oil. Evaporator temperature and flow rate were optimized using response surface methodology and two models were obtained for the FFA content left and loss of tocopherols. An applicable parameter zone was created to obtain a certain FFA (0.5% for example) content. In general, conditions that result in a lower FFA content will lead to a higher loss of tocopherols. In most parts of the parameter zone, 50% loss of tocopherols will be expected. The deodorization study of randomized SL from fish oils and tricaprin indicated that SPD in comparison with batch deodorization gave a product of a poorer sensoric quality.     

Purification of tocopherols and phytosterols by a two-step <Emphasis Type="Italic">in situ</Emphasis> enzymatic reaction

The purification of tocopherols and phytosterols (referred to as sterols) from soybean oil deodorizer distillate (SODD) was attempted. Tocopherols and sterols in the SODD were first recovered by short-path distillation, which was named sODD tocopherol/sterol concentrate (SODDTSC). The SODD-TSC contained MAG, DAG, FFA, and unidentified hydrocarbons in addition to the two substances of interest. It was then treated with Candida rugosa lipase to convert sterols to FA steryl esters, acylglycerols to FFA, and FFA to FAME. Methanol (MeOH), however, inhibited esterification of the sterols. Hence, a two-step in situ reaction was conducted: SODDTSC was stirred with 20 wt% water and 200 U/g mixture of C. rugosa lipase at 30°C, and 2 moles of MeOH per mole of FFA was added to the reaction mixture after 16h. The lipase treatment for 40 h in total achieved 80% conversion of the initial sterols to FA steryl esters, complete hydrolysis of the acylglycerols, and a 78% decrease in the initial FFA content by methyl esterification. Tocopherols did not change throughout the process. To enhance the degree of steryl and methyl esterification, the reaction products, FA steryl esters and FAME, were removed by short-path distillation, and the resulting fraction containing tocopherols, sterols, and FFA was treated with the lipase again. Distillation of the reaction mixture purified tocopherols to 76.4% (recovery, 89.6%) and sterols to 97.2% as FA steryl esters (recovery, 86.3%).     

Lipase‐catalysed production and chemical composition of diacylglycerols from soybean oil deodoriser distillate

Diacylglycerols (DAG) were enzymatically produced by lipase‐catalysed esterification of glycerol with fatty acids from soybean oil deodoriser distillate (SODD). Effects of reaction parameters such as reaction time, temperature, enzyme type, enzyme load, substrate molar ratio and water content, as well as the effect of molecular sieves as water adsorbent were studied. Lipozyme RM IM was determined to be the most effective among the lipases screened. The following conditions yielded 69.9% DAG (all percentages are wt/wt): 4 h reaction time, 65 °C reaction temperature, 10% Lipozyme RM IM, 2.5:1 fatty acid to glycerol molar ratio, and 30% molecular sieves. DAG synthesis of 11.9% was still observed at 10% water content. After purification, the product oil contained 86.3% DAG. This oil consisted predominantly of 1,3‐diolein (19.1%), 1‐oleoyl‐3‐linoleoyl‐glycerol (18.2%) and 1‐oleoyl‐2‐linoleoyl‐glycerol (16.6%). The fatty acid profile of the oil was similar to that of refined, bleached and deodorised (RBD) soybean oil. The % ratio of 1,3‐ to 1,2‐positional isomers of DAG was at 56:44.