Formation of conjugated linoleic acids in soybean oil during hydrogenation with a nickel catalyst as affected by sulfur addition

The effects of sulfur addition on the formation of conjugated linoleic acid (CLA) isomers were studied during the hydrogenation of soybean oil with a nonselective type nickel catalyst. Sulfur addition greatly promoted CLA formation in soybean oil during hydrogenation. As the amount of sulfur increased to a certain level, the maximal quantity of CLA in soybean oil during hydrogenation increased greatly. However, further increase in sulfur addition above the certain level decreased CLA formation. The optimal sulfur level for the promotion of CLA formation differed greatly with the amount of nickel used. It was of great interest to find that the optimal ratio of sulfur to nickel for the promotion of CLA formation was always 0.06:1, regardless of the nickel amount used. At the same ratio of sulfur to nickel, higher nickel content induced significantly higher production of CLA (p < 0.05). At the optimal sulfur to nickel ratio, an increase in the nickel amount from 0.05 to 0.15% produced approximately 1.5 times higher levels of CLA during hydrogenation under the tested conditions. The CLA isomer compositions were greatly affected by both sulfur addition and amounts of nickel used for treatment. This is the first report of the possibility that the total quantity of CLA and their isomer composition could be manipulated during hydrogenation by controlling the amounts of sulfur and nickel.     

Influence of feeding soybean oil on conjugated linoleic acid content in beef

Forty-eight steers were used to study the influence of feeding soybean oil (SO) on the conjugated linoleic acid (CLA) content of beef. Steers were fed either a control diet containing 954 g/kg of dry matter (DM) corn-based concentrate (CTL) or a control diet supplemented with SO at 20 (SO2) or 40 g/kg (SO4) of diet DM for 105 days. Adipose tissue samples were collected from the M. longissimus dorsi (LD) and from the M. semitendinosus (ST) on days 0 and 63 of the experiment. Adipose and muscle tissue samples were collected from the LD and ST immediately after slaughter. Feeding 40 g/kg of DM as SO increased the proportions of trans-C(18:1) in beef lipid as compared to CTL and SO2 treatments. The C(18:2) cis-9, trans-11 isomer of CLA as a proportion of total fat was not different in adipose and muscle across treatments. Supplementing SO increased C(18:2) trans-10, cis-12 CLA in adipose tissue of the LD. Supplementing high-grain finishing diets with SO is not an effective strategy to enhance the C(18:2) cis-9, trans-11 isomer of CLA in beef.     

Optimization of production of conjugated linoleic acid from soybean oil

Linoleic acid from soybean oil was used to synthesize conjugated linoleic acid (CLA), and the response surface methodology (RSM) was applied to optimize the process. A temperature of -35 degrees C and a solvent to oil sample ratio of 8 were suggested for removal of saturated fatty acids by low-temperature crystallization. The ratio of oil sample/urea/methanol suggested was 1:2:5.5 (w/w/v) for removal of oleic acid by urea crystallization. A temperature of 150 degrees C and a time of 140 min were found to be the optimal conditions in the isomerization for the production of c-9,t-11 and t-10,c-12 CLA isomers.     

Conjugated linoleic acid isomers in partially hydrogenated soybean oil obtained during nonselective and selective hydrogenation processes

Partially hydrogenated soybean oil samples were collected during selective and nonselective hydrogenation processes. The formation of conjugated linoleic acids (CLAs) during hydrogenation was greatly dependent on the types and duration of hydrogenation processes. During hydrogenation processes, CLA contents increased initially. After reaching maximum CLA content, the content decreased during hydrogenation. Selective hydrogenation was much more favorable for the formation of conjugated linoleic acids. With nonselective hydrogenation process, the total CLA content was a maximum (9.06 mg total CLA/g oil) at 35 min. However, with the selective hydrogenation process, the total CLA content was a maximum (98.27 mg total CLA/g oil) at 210 min. The CLA contents in some of the tested selectively hydrogenated soybean oils were among the highest ever reported in foods.     

Free radical scavenging properties of conjugated linoleic acids.

Conjugated linoleic acids (CLA) were investigated for free radical scavenging properties against the stable 2,2-diphenyl-1-picryhydrazyl radical (DPPH.) by electron spin resonance (ESR) spectrometry and spectrophotometric methods. ESR results demonstrated that CLA directly reacted and quenched free DPPH radicals in benzene, while spectrophotometric analysis showed the radical scavenging capacity of CLA in ethanol. Dose and time effects of CLA-DPPH. reactions were observed in both tests. The ED(50) of CLA was 18 mg/mL under experimental conditions. CLA are much weaker radical scavengers as compared to vitamin E, vitamin C, and BHT. Kinetics of CLA-DPPH. reactions was different to that of linoleic acid (LA)-DPPH. reactions. CLA reacted and quenched DPPH radicals at all tested levels without a lag phase, while LA had a lag phase and showed no radical quenching activity at levels of 5-80 mg/mL in 30 min. These data indicated that CLA can provide immediate protection against free radicals, but LA cannot.     

Identification of conjugated linoleic acids in hydrogenated soybean oil by silver ion-impregnated HPLC and gas chromatography-ion impacted mass spectrometry of their 4,4-dimethyloxazoline derivatives

Hydrogenated soybean oil was obtained after 10 min of hydrogenation with 0.5% selective type Ni catalyst at 230 degrees C, a hydrogenation pressure of 0.049 MPa, and an agitation rate of 300 rpm. The conjugated linoleic acid isomers in the hydrogenated soybean oil were isolated by using a silver ion-impregnated HPLC. Gas chromatography-mass spectrometry of 4,4-dimethyloxazoline (DMOX) derivatives of the isolated conjugated linoleic acid isomers were carried out for the identification of their chemical structures. By interpreting the mass spectra of the DMOX derivatives of conjugated linoleic acid isomers isolated by silver ion-impregnated HPLC, 20 different conjugated linoleic acid isomers present in hydrogenated soybean oil were identified. This is the first report for the mass spectrometric identification of the conjugated linoleic acid isomers present in hydrogenated vegetable oil.     

Effects of rhodium heterogeneous catalyst and isomerization conditions on linoleic acid conjugation of soybean oil

Rhodium heterogeneous catalyst was used to catalyze isomerization of linoleic acid in soybean oil to conjugated linoleic acid (CLA). A central composite rotatable design with five levels of three variables, namely, reaction temperature, stirring speed, and reaction time, was used to determine the maximum CLA yield. The formation of CLA during isomerization was greatly dependent on the reaction temperature and time. The CLA content of soybean oil increased from 0.63 to 202.42 mg/g oil when isomerization was done at 200 degrees C, with a stirring speed of 200 rpm for 49 min. Analysis of triacylglycerol positions showed that linoleic acid at any position in a triacylglyceride could possibly be isomerized to CLA.     

Effect of extrusion parameters on conjugated linoleic acids of corn extrudates

The effects of extrusion temperature, 150-190 degrees C, and torque, 50-70%, on the content and configuration of conjugated linoleic acids (CLA) in corn extrudates were analyzed by GC and HPLC. At a temperature of 150 degrees C, CLA content increased from 1.2 mg/g of oil in feed to 7.8 mg/g of oil in corn extrudates. A decrease in total CLA (P < 0.05) was obtained when the product temperature was further increased to 190 degrees C. Alteration of CLA geometrical configuration was observed at higher extrusion temperatures. trans,trans-CLA significantly increased (P < 0.05) from 10.2% in feed to 11.9% of CLA at the extrusion condition of 190 degrees C and 70% torque. The highest expansion of extrudates was found at the product temperature of 150 degrees C and 70% torque. This extrusion condition also gave the maximum total CLA content and minimum trans,trans-CLA formation.     

Photocatalytic production and processing of conjugated linoleic acid-rich soy oil

Daily intake of conjugated linoleic acid (CLA), an anticarcinogenic, antiatherosclerotic, antimutagenic agent, and antioxidant, from dairy and meat products is substantially less than estimated required values. The objective of this study was to obtain CLA-rich soybean oil by a customized photochemical reaction system with an iodine catalyst and evaluate the effect of processing on iodine and iodo compounds after adsorption. After 144 h of irradiation, a total CLA yield of 24% (w/w) total oil was obtained with 0.15% (w/w) iodine. Trans,trans isomers (17.5%) formed the majority of the total yield and are also associated with health benefits. The isomers cis-9,trans-11 and trans-10,cis-12 CLA, associated with maximum health benefits, formed approximately 3.5% of the total oil. This amount is quite significant considering that total CLA obtained from dairy sources is only 0.6%. ATR-FTIR, 1H NMR, and GC-MS analyses indicated the absence of peroxide and aldehyde protons, providing evidence that secondary lipid oxidation products were not formed during the photochemical reaction. Adsorption processing vastly reduced the iodine and iodocompounds without CLA loss. Photocatalysis significantly increased the levels of CLA in soybean oil.     

Effect of the degree of processing on soy oil conjugated linoleic acid yields

Photoirradiation processing can be used to rapidly synthesize conjugated linoleic acid (CLA) in large quantities in soy oil. The objective of this study was to evaluate the effect of the level of refining of soy oil on CLA yields and oxidative properties after photoirradiation. Crude, alkali-refined, alkali-refined bleached, and alkali-refined bleached and deodorized (RBD) soy oils were photoirradiated in a pilot-plant processing system for 12 h with 0.35% iodine catalyst at 47 degrees C. RBD soy oil gave the highest total CLA yield of 16.3% of total oil with 4.3% cis, trans- and trans, cis-CLA isomers. Oxidative stability as measured by weight gain during incubation at 64 degrees C showed that iodine decreased the induction time of soy oil samples by 2-4 days. Photoirradiation processing further decreased the induction time by 2 days as a result of loss of total tocopherols. Iodine addition increased the titratable acidity in all the samples of soy oil. However, the level of refining affects this increase of titratable acidity, and RBD soy oil was found to be the most stable. The study indicates that RBD soy oil was the most suitable candidate for photoirradiation processing. Thus, soy oil should be alkali-refined, bleached, deodorized, and then photoprocessed followed by a secondary adsorption step to remove the iodine catalyst to obtain a RBD CLA-rich soy oil.     

Effect of antioxidants on soy oil conjugated linoleic acid production and its oxidative stability

Conjugated linoleic acid (CLA)-rich soy oil can be produced by photoisomerization of soy oil linoleic acid to produce a soy oil with up to 20% CLA. Recent studies indicate that mixed soy tocopherols added to refined bleached deodorized (RBD) oil produced significant increase in soy CLA yield during soy oil linoleic acid photoisomerization. However, the effect of common synthetic free radical scavenging antioxidants and specific tocopherols on CLA yield and its oxidative stability is not known. Therefore, this investigation evaluated the effects of various antioxidant systems on soy oil CLA yield and oxidative stability. Soy oil with added antioxidants consisting of combinations of mixed tocopherols (MT), ascorbyl palmitate (AP), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ) was photoisomerized to produce CLA- rich soy oil. The CLA content was determined by GC-FID analysis and oxidative stability by peroxide value (PV). The soy oil in the presence of TBHQ, MT alone and MT with 500 ppm of AP produced significantly greater CLA yields and improved oxidative stability compared to a control without added antioxidants (p < 0.05). However, added mixed tocopherols produced the greatest CLA yield and also reduced PV relative to the control. Tocopherols in the form of α-, γ- and δ-tocopherols were then each examined as to their relative effect on CLA yields and PV. The largest increase in CLA yield was obtained with 1800 ppm of γ-tocopherols with reduced PV. Mixed tocopherols, TBHQ and γ-tocopherols can be used to increase CLA yield and reduce PV of soy oil during linoleic acid photoisomerization.     

Determination of the conjugated linoleic acids in cow's milk fat by Fourier transform Raman spectroscopy

The collective term "conjugated linoleic acid" or "CLA" generally refers to a mixture of conjugated positional and geometric isomers of linoleic (cis-9,cis-12-octodecadienoic) acid. In nature, these isomers are mainly formed in the rumen by biohydrogenation of polyunsaturated fatty acids. This study concerns a first trial of CLA determination in cow's milk fat by Raman spectroscopy. The spectra of pure cis-9-oleic, cis-9,cis-12-linoleic, cis-9,trans-11-linoleic, and trans-10,cis-12-linoleic acids have been examined in comparison with the spectra of selected milk-fat samples containing between 0 and 3% of CLA. The trial of CLA determination by Raman spectroscopy on cow milk fat has reached its objective with the two following results. First, the examination of the Raman spectra allows to identify three specific Raman signals of the chemical bonds associated to the cis,trans conjugated C=C in the rumenic and trans-10,cis-12-octodecadienoic acids at 1652, 1438, and 3006 cm(-1). Second, the calibration of Raman spectrometer for the CLA determination has indicated that these three specific signals suit very well for the accurate and reliable measurement of CLA concentration in milk fat. To our knowledge, the present study is the first successful attempt to determine the CLA content of milk fat by a spectrophotometric method.     

Effect of conjugated linoleic acids on the activity and mRNA expression of 5- and 15-lipoxygenases in human macrophages

Lipoxygenases are a family of non-heme enzyme dioxygenases. The role of lipoxygenases is synthesis of hydroperoxides of fatty acids, which perform signaling functions in the body. Studies on conjugated linoleic acids (CLAs) as fatty acids with a potential anti-atherosclerotic function have recently been initiated. The aim of the study was to test the effect of CLAs and linoleic acid on 5- and 15-lipoxygenase (5-LO, 15-LO-1) enzyme activity, their mRNA expression, and concentration in the cells. It was also desired to determine whether the CLAs are substrates for the enzymes. For the experiments monocytic cell line (THP-1) and monocytes obtained from human venous blood were used. Monocytes were differentiated to macrophages: THP-1 (CD14+) by PMA administration (100 nM for 24 h) and monocytes from blood (CD14+) by 7-day cultivation with the autologous serum (10%). After differentiation, macrophages were cultured with 30 microM CLAs or linoleic acid for 48 h. The 15- and 5-lipoxygenase products were measured by HPLC method. mRNA expression and protein content were analyzed by real-time PCR and Western blot analysis. The in vitro studies proved that both CLA isomers are not substrates for 15-LO-1; in ex vivo studies hydroxydecadienoic acid (HODE) concentration was significantly reduced (p = 0.019). The trans-10,cis-12 CLA isomer reduced HODE concentration by 28% (p = 0.046) and the cis-9,trans-11 CLA isomer by 35% (p = 0.028). In macrophages obtained from THP-1 fatty acids did not change significantly mRNA expression of the majority of the investigated genes. CLAs did not change the content of 5-LO and 15-LO-1 proteins in macrophages obtained from peripheral blood. Linoleic acid induced 15-LO-1 expression (2.6 times, p < 0.05). CLAs may perform the function of an inhibitor of lipoxygenase 15-LO-1 activity in macrophages.     

Performance comparison of UV and FT-Raman spectroscopy in the determination of conjugated linoleic acids in cow milk fat

The determination of conjugated linoleic acids (CLA) in cow milk fat was studied by using UV (210-250 nm) and Fourier transform (FT)-Raman (900-3400 cm (-1)) spectroscopy in order to determine the best spectrophotometric technique for routine analysis of milk fat. A collection of 57 milk fat samples was randomly divided into two sets, a calibration set and a validation set, representing two-thirds and one-third of the samples, respectively. All calculations were performed on the calibration set and then applied to the validation set. The CLA content ranged from 0.56 to 4.70%. A comparison of various spectral pretreatments and different multivariate calibration techniques, such as partial least-squares (PLS) and multiple linear regression (MLR), was done. This paper shows that UV spectroscopy is as reliable as FT-Raman spectroscopy to monitor CLA in cow milk fat. The best calibration for FT-Raman was given by a PLS model of seven factors with a standard error of prediction (SEP) of 0.246. For UV spectroscopy, PLS models were also better than MLR models. The most robust PLS model was constructed with only one factor and with SEP=0.288.     

Antioxidant evaluation and oxidative stability of structured lipids from extravirgin olive oil and conjugated linoleic acid

Structured lipid (SL) was synthesized from extravirgin olive oil (EVOO) and conjugated linoleic acid (CLA) via a lipase-catalyzed reaction. CLA provides a variety of health benefits, but it is not consumed in free fatty acid form. The synthesized SL olive oil contained 42.5 mol % CLA isomers, and the major isomers were cis-9,trans-11-CLA (16.9 mol %) and trans-10,cis-12-CLA (24.2 mol %). The antioxidant activity determined by the radical scavenging capacity with the 2,2-diphenyl-1-picrylhydrazyl radical was lower in SL olive oil than in EVOO. The oxidative stability was also lower in SL olive oil since it had a higher peroxide value, rho-anisidine value, and 2-thiobarbituric acid reactive substances values during 20 days of storage at 60 degrees C. This observation could be due to the reduction in the natural phenolic compounds (97%) and tocopherols (56%), and the incorporated CLA with two conjugated double bonds in the SL olive oil. The oxidative stability of SL olive oil was increased by added rosemary extracts at concentrations of 100, 200, and 300 ppm. The present study suggests that the SL olive oil may be a suitable way to incorporate or deliver CLA into human diets. However, the addition of a proper antioxidant would be required for improving its oxidative stability.     

黑土农田大豆产量形成过程的模拟验证

以中国科学院海伦农业生态站长期定位水肥耦合试验数据为依据,模拟大豆产量形成过程。首先建立大豆品种遗传属性数据库和相应的模型参数,利用DSSAT模型系列中的CropGro-Soybean模型,对大豆品种遗传属性、作物产量和生长过程中土壤水分进行了模拟验证。模拟结果表明,CropGro-Soybean模型能够准确地模拟大豆生育期,相对误差在-2％～3％之间,均方根误差RMSE为2．3。对不同年际不同田间处理的大豆产量模拟结果分析的相对误差在-7％～9％之间,均方根误差RMSE为75．9,模型性能指数EF为0．8。模型对不同层次土壤水分变化的模拟效果也较好。     

Isomeric distribution of conjugated linoleic acids (CLA) in the tissues of layer hens fed a CLA diet

The isomeric distribution of conjugated linoleic acids (CLA) in the tissue lipids of hens in relation to that in the diet was examined. Silver-ion high-performance liquid chromatography was used to quantify individual CLA isomers in total tissue lipids, phospholipids, and triacylglycerols. It was found that the deposition of CLA isomers in hen tissues was selective. All tissues including serum, liver, heart, kidney, abdominal fat, and leg and breast muscles had lesser amounts of total cis/trans isomers ranging from 75.87 to 89.13% of total CLA, which was in contrast to the value of 92% of total CLA in the dietary lipids. Total trans/trans isomers in all tissue lipids ranging from 6.11 to 18.02% of total CLA were greater than that in the diet (4.19%). Among the individual trans/trans isomers, all tissues except for adipose tissue and brain incorporated greater amounts of t-12,t-14-18:2, t-11,t-13-18:2,t-10,t-12-18:2, t-9,t-11-18:2, and t-18,t-10-18:2 compared with the values of the diet. Within the cis/trans group, lesser amounts of c-10,t-12/t-10,c-12-18:2 were found to incorporate into all tissues compared with the value of the diet. Serum and liver had higher percentages of c-9,t-11/t-9,c-11, whereas the other tissues had similar levels of this isomer compared with that of the diet. It was also observed that supplementation of CLA in the diet of layer hens decreased the concentration of docosahexaenoic acid (22:6n-3) in all of the tissue lipids. It is concluded that dietary CLA can transfer to the tissue but that incorporation of CLA isomers into the tissue is selective in hens.     

Effects of conjugated linoleic acid (CLA) isomers on oxygen diffusion-concentration products in liposomes and phospholipid solutions

Conjugated linoleic acids (CLAs) are a group of octadecadienoic acids (18:2) that are naturally present in food products and may have beneficial health effects. Liposomes and ethanol solutions were prepared by mixing synthetic phosphatidylcholines (PCs) with c9,t11-CLA, t10,c12-CLA, and linoleic acid (LA) in the sn-2 position into natural PCs from soybean, egg yolk, rat brain, and rat heart at 5 mol %. The oxygen diffusion-concentration products were measured using electron spin resonance spin-label oximetry methods. Individual synthetic PCs, the phospholipid matrix, and the tested lipid systems all exhibited influence on oxygen diffusion-concentration products during lipid peroxidation. Incorporating 5 mol % PC(c9,t11-CLA) into soy and egg yolk PC increased oxygen consumption in liposome suspensions while it was decreased in rat heart and brain PCs. On the other hand, PC(t10,c12-CLA) increased oxygen consumption in mixtures with egg yolk and rat heart PC but decreased it in soybean and rat brain PC. By comparison, PC(LA) decreased oxygen consumption in every case. In ethanol solutions, all of the synthetic PCs suppressed the capacity to generate peroxide radicals in the order of LA > c9,t11-CLA > t10,c12-CLA. In addition, PCs containing individual CLA isomers and LA differed in their capacities to react with and quench DPPH radicals in both ethanol solution and liposome, suggesting differences between CLA isomers and LA in DPPH radical-fatty acid interactions. Incorporation of CLA isomers and LA into dimyristyl-PC reduced the phase transition temperature from 23.6 to 23.1 and 23.3 degrees C, respectively. The results of this study provide evidence that the behavior of CLA isomers differs in the microenvironment of membranes possibly due to structural differences that affect the permeability of membranes to oxygen and lipid peroxidation.     

Dietary combination of conjugated linoleic acid (CLA) and pine nut oil prevents CLA-induced fatty liver in mice

Conjugated linoleic acid (CLA) strongly prevents fat accumulation in adipose tissue of mice, even if hepatic fat deposition and insulin resistance are concomitantly observed. This study investigated the possibility of maintaining the antiadiposity properties of CLA while preventing adverse effects such as liver steatosis and hyperinsulinemia. To this end, mice were divided into three groups and fed a standard diet (control) or a diet supplemented with 1% CLA (CLA) or a mixture of 1% CLA plus 7.5% pine nut oil (CLA + P). The combination of CLA + P preserved the CLA-mediated antiadiposity properties (70% fat reduction), preventing hepatic steatosis and a sharp increase in plasmatic insulin starting from the eighth week of CLA treatment. The assay of both fatty acid synthesis and oxidation in the CLA + P mice revealed a time-dependent biphasic behavior of the corresponding enzymatic activities. A sudden change in these metabolic events was indeed found at the eighth week. A strong correlation between the changes in key enzymes of lipid metabolism and in insulin levels apparently exists in CLA-fed mice. Furthermore, lower levels of lipids, in comparison to values found in CLA-fed mice, were observed in the liver and plasma of CLA + P-fed animals.