Supercritical carbon dioxide extraction (SC-CO2) technology was used to extract oil from Eucommia ulmoides seed. The optimum conditions and significant parameters in SC-CO2 were obtained using response surface methodology (RSM). The qualities of the extracted oil were evaluated by physicochemical properties, fatty acid composition, vitamin E composition. It was found that the optimum extraction parameters were at pressure of 37 MPa, temperature of 40°C, extraction time of 125 min and CO2 flow rate of 2.6 SL/min. Pressure, temperature and time were identified as significant parameter effecting on extraction yield. The importance of evaluated parameters decreased in the order of pressure > extraction time > temperature > CO2 flow rate. GC analysis indicated that E. ulmoides seed oil contained about 61% of linolenic acid and its fatty acid composition was similar with that of flaxseed oil and perilla oil. The content and composition of vitamin E was determined using HPLC. The E. ulmoides seed oil was rich in vitamin E (190.72 mg/100 g), the predominant vitamin E isomers were γ- tocopherol and δ- tocopherol, which accounted for 70.87% and 24.81% of the total vitamin E, respectively. The high yield and good physicochemical properties of extracted oil support the notion that SC-CO2 technology is an effective technique for extracting oil from E. ulmoides seed.
High-Performance Liquid Chromatography (HPLC) methods via evaporative light scattering (ELS) and refractive index (RI) detectors are used by the local palm oil industry to monitor the TAG profiles of palm oil and its fractions. The quantitation method used is based on area normalization of the TAG components and expressed as percentage area. Although not frequently used, peak-area ratios based on TAG profiles are a possible qualitative method for characterizing the TAG of palm oil and its fractions. This paper aims to compare these two detectors in terms of peak-area ratio, percentage peak area composition, and TAG elution profiles. The triacylglycerol (TAG) composition for palm oil and its fractions were analysed under similar HPLC conditions i.e. mobile phase and column. However, different sample concentrations were used for the detectors while remaining within the linearity limits of the detectors. These concentrations also gave a good baseline resolved separation for all the TAGs components. The results of the ELSD method’s percentage area composition for the TAGs of palm oil and its fractions differed from those of RID. This indicates an unequal response of TAGs for palm oil and its fractions using the ELSD, also affecting the peak area ratios. They were found not to be equivalent to those obtained using the HPLC-RID. The ELSD method showed a better baseline separation for the TAGs components, with a more stable baseline as compared with the corresponding HPLC-RID. In conclusion, the percentage area compositions and peak-area ratios for palm oil and its fractions as derived from HPLC-ELSD and RID were not equivalent due to different responses of TAG components to the ELSD detector. The HPLC-RID has a better accuracy for percentage area composition and peak-area ratio because the TAG components response equally to the detector.
The intake of trans fatty acids (TFAs) in foods changes the ratio of low density lipoprotein (LDL) to high density lipoprotein (HDL) cholesterol in blood, which causes cardiovascular disease. TFAs are formed by trans isomerization of unsaturated fatty acids (UFAs). The most recognized formation mechanisms of TFAs are hydrogenation of liquid oil to form partially hydrogenated oil (PHO,) and biohydrogenation of UFAs to form TFA in ruminants. Heating oil also forms TFAs; however, the mechanism of formation, and the TFA isomers formed have not been well investigated. In this study, the trans isomerization mechanism of unsaturated fatty acid formation by heating was examined using the model compounds oleic acid, trioleate, linoleic acid, and trilinoleate for liquid plant oil. The formation of TFAs was found to be suppressed by the addition of an antioxidant and argon gas. Furthermore, the quantity of formed TFAs correlated with the quantity of formed polymer in trioleate heated with air and oxygen. These results suggest that radical reactions form TFAs from UFAs by heating. Furthermore, trans isomerization by heating oleic acid and linoleic acid did not change the original double bond positions. Therefore, the distribution of TFA isomers formed was very simple. In contrast, the mixtures of TFA isomers formed from PHO and ruminant UFAs are complicated because migration of double bonds occurs during hydrogenation and biohydrogenation. These findings suggest that trans isomerization by heating is executed by a completely different mechanism than in hydrogenation and biohydrogenation.
In this study, we measured the quantity of marine-derived tocopherol (MDT), a monounsaturated vitamin E (VE), stored in the body tissue of mice fed with a diet containing a VE-rich fraction extracted from salmon roe. We first prepared the calibration curves for the MDT concentration using an HPLC-fluorescence system. Ranging from 0.016 to 50 μg/mL, the slope was expressed as first-order equations, with R2 values = 0.99. The mice were fed with an AIN-93 based diet containing MDT in doses of 21.4 mg/kg for 4 weeks, and the storage in the heart, lung, liver, stomach, small intestine, large intestine, kidney, pancreas, spleen, testis, skeletal muscle, visceral white adipose tissue (WAT), subcutaneous WAT and brain was quantified. MDT was widely distributed in tissues throughout the whole body, with higher accumulations observed in the adipose tissue, liver and kidney. These results demonstrate means to estimating the MDT concentration in natural products and in the bodies of animals and contribute to the understanding of the physiological functions of MDT in relation to human health.
An improved colorimetric method for determination of cyanide content in bitter almond oil was developed. The optimal determination parameters were as follows: volume ratio of hydrochloric acid to bitter almond oil (v/v), 1.5:1; holding time for hydrolysis, 120 min; and volume ratio of distillation solution to bitter almond oil (v/v), 8:1. Analytical results showed that the relative standard deviations (SDs) of determinations were less than 10%, which satisfies the test requirements. The results of high-performance liquid chromatography and measurements exhibited a significant correlation (R = 0.9888, SD = 0.2015). Therefore, the improved colorimetric method can be used to determine cyanide content in bitter almond oil.
Ion-pair amphiphiles (IPAs, also known as catanionic surfactants) are lipid-like double-chained molecules potentially used for fabricating liposome-like vesicular drug and gene carriers. Frequently ethanol and cholesterol are added to modulate the properties of their bilayer membranes. Effects of ethanol and cholesterol on the fundamental properties of IPA bilayers such as thermotropic phase behavior, however, is not known. In this work, the bilayer phase transition behavior of two IPAs (decyltrimethylammonium-tetradecyl sulfate, DeTMA-TS, and dodecyltrimethylammonium-dodecyl sulfate, DTMA-DS) in tris buffer with various amounts of ethanol was studied by using differential scanning calorimetry (DSC). Effect of cholesterol (CHOL) addition on bilayer phase transition of IPAs with 20 vol% ethanol was thereafter systematically investigated. The experimental results showed that the main phase transition temperature (Tm) was monotonously decreased with the increase of ethanol concentration up to 30 vol%. The degree of Tm depression by ethanol is essentially the same for the two IPAs regardless of different symmetry in the hydrocarbon chains. Further addition of CHOL, however, caused a slight decrease in Tm on the one hand and a significant decrease in the enthalpy of phase transition on the other hand. When the added CHOL exceeded a specific amount, the phase transition disappeared. More hasty disappearance of phase transition was found for IPA with asymmetric structure than the symmetric one. Possible mechanisms of ethanol effect based on binding in the headgroup region of the bilayers and CHOL effect based on opposite (condensing and disordering) interactions with IPA molecules in bilayers, respectively, were proposed.
A new emulsification technique using porous silica particles was studied as a facile and instantaneous formation method for thermodynamically unstable emulsions. In this study, oil encapsulated in silica particles was released instantly upon the addition of a phosphate buffer, forming an oil-in-water (O/W) emulsion. Emulsion formation was inhibited or promoted using lipophilic or hydrophilic surfactant additives, respectively. We concluded that this phenomenon is affected by the wettability of the soybean oil on the silica surface, which is controlled by the surfactant. We prepared submicron size emulsions in a simple method involving the addition of the silica particles to an aqueous solution. This spontaneous emulsification technique could be applied to the formation of solid-in-oil-in-water (S/O/W) emulsions for oral delivery of hydrophilic peptide medicine.
Adding fatty acids to an oil-in-water (O/W) emulsion changes the stability of the emulsion. In this study, we prepared a series of O/W emulsions consisting of oil (triolein/fatty acid mixture), water and a range of surfactants (sucrose fatty acid esters) with varying hydrophilic–lipophilic balance (HLB) in order to determine the effects of alkyl chain length and the degree of unsaturation of the fatty acid molecules on the stability of the emulsions. As a result, sucrose fatty acid esters with HLB = 5–7 were suitable for obtaining O/W emulsions. In addition, the creaming phenomenon was inhibited for 30 days or more when fatty acids having a linear saturated alkyl chain with 14 or more carbon atoms were added. These findings are useful for designing stable O/W emulsions for food and cosmetic products.
The effects of four detergent enzymes on the Orange II decoloration reaction by horseradish peroxidase (HRP) were investigated. Stainzyme, Lipex, Celluclean, and Savinase did not affect the decoloration reaction up to a concentration of 0.1% Under weak alkaline to alkaline conditions, the presence of four enzymes did not affect the decoloration reaction, but affected the decoloration reaction under neutral to acidic conditions. The effect of Ca2+ and Mg2+ concentrations on Orange II decoloration reactions in the presence of four enzymes was also investigated. Ca2+ and Mg2+ did not affect the decoloration reaction up to 500 ppm.
Fatty acid methyl esters were synthesized from palm fatty acid distillate (PFAD) and methanol in a packed bed reactor via lipase-catalyzed esterification. The PFAD consisted of 91 wt% of free fatty acids, 2 wt% monoacylglycerides, 3 wt% diacylglycerides, and 4 wt% triacylglycerides. t-Butanol was employed as a reaction medium and a mixed enzyme consisting of Lipozyme TL IM from Thermomyces lanuginosus and Novozym 435 from Candida antarctica was employed as the biocatalyst. The effect of mixed enzyme was investigated and the optimum blending ratio (w/w) of Novozym 435 to Lipozyme TL IM was 5:95. Using the mixed enzyme, the optimum molar ratio (PFAD to methanol) and temperature were determined to be 1:6 and 30°C, respectively. Under the optimized conditions, the maximum yield of ca. 96% was achieved.
The biological activities of phospholipids (PLs) have attracted people’s attention, especially marine phospholipids with omega-3 polyunsaturated fatty acids DHA and EPA. In this study, we investigated the immunity activation of macrophages in vitro by phospholipids from skipjack brain. The phospholipids were extracted with hexane and ethanol ultrasonication instead of the traditional method of methanol and chloroform. The content of phospholipids from Skipjack brain was 19.59 g/kg by the method (the ratio of hexane and ethanol 2:1, 40 min, 35°C, 1:9 of the ratio of material to solvent, ultrasonic power 300W, ultrasonic extraction 2 times). The RAW264.7 macrophages were stimulated by the phospholipids from the Skipjack, by which the volume, viability and phagocytosis of macrophages were increased. The concentration of NO and the activity of SOD of the cells were also enhanced. The gene expressions of IL-1β, IL-6, iNOS and TNF-α mRNA assayed by RT-PCR were up-regulated. Phospholipids from brain of Skipjack Tuna could activate macrophages immunity which displayed to induce pro-inflammatroy cytokines mRNA expression.
Ulcerative colitis is a well-known inflammatory bowel disease. Although there are drugs that are effective against this disease, the prevention and attenuation of ulcerative colitis by food rich in functional ingredients without side effects is desired because some drugs have side effects. In this study, we investigated the effects of yuzu (Citrus junos Tanaka), a citrus fruit native to northeast Asia, on a mouse dextran sulfate sodium (DSS)-induced colitis model. Mice given drinking water containing DSS showed significant weight loss, colon shortening, diarrhea, and visible fecal blood. In contrast, mice fed a diet containing 5% yuzu peel for 14 d before receiving DSS showed significant attenuation of these phenotypes. To clarify the mechanism underlying the attenuation, we investigated the anti-inflammatory and antioxidant effects of yuzu peel. We found that yuzu peel extract suppressed tumor necrosis factor-α (TNF-α) production in lipopolysaccharide (LPS)-stimulated mice and murine macrophage cell line through suppression of nuclear factor-κB (NF-κB) activation. In addition, we confirmed that yuzu peel extract had a moderate antioxidant effect. These results suggest that yuzu peel attenuates the pathologies of DSS-induced colitis by coordinately suppressing inflammation and oxidative stress against lipids in vivo.
The aim of the present research was to investigate the effect of fish oil, crude Nigella sative oil and combined fish oil/Nigella sative volatile oil as hepato-regenerative and renal protective supplements. The oils were administered as emulsions to rat model with liver injury induced by CCl4. Plasma activities of transaminases (AST and ALT) were evaluated as liver function indicators, while plasma creatinine and urea and creatinine clearance were determined as markers of kidney function. Plasma malondialdehyde (MDA), nitrite (NO) and tumor necrosis factor-α (TNF-α) were estimated to assess the exposure to oxidative stress and subsequent inflammation. Liver fat was extracted and their fatty acids´ methyl esters were determined using gas chromatography. Results showed that plasma activities of AST and ALT were significantly higher in CCl4 control group compared to control healthy group. Plasma levels of creatinine and urea increased significantly in CCl4 control, while creatinine clearance was reduced significantly in the same group. All rat treated groups given the three oil emulsions showed improvement in liver function pointing to the initiation of liver regeneration. The combination of fish oil/Nigella sative volatiles showed the most promising regenerative activity. Oxidative stress and inflammation which were increased significantly in CCl4 control group showed improvement on administration of the three different oil emulsions. Fatty acids methyl ester of liver fat revealed that rats treated with fish oil/Nigella sative volatile oil presented the highest content of unsaturated fatty acids (45.52% ± 0.81) while fish oil showed the highest saturated fatty acids (53.28% ± 1.68). Conclusion; Oral administration of oil emulsions of native fish oil, Nigella sative crude oil and combined fish oil/Nigella sative volatile oil reduced liver and kidney injury in rat model of CCl4 through exerting anti-inflammatory and antioxidant activity. Fish oil/Nigella sative volatile oil emulsion was the most promising hepato-regenerative and reno-protective formula among the different groups.
A new method for utilization of hydrogel is proposed here for the preparation of solid base catalysts for the transesterification of vegetable oil. When a solution of KF is mixed with a solution of Ca(NO3)2, CaF2 is obtained and inactive as a catalyst in the transesterification of vegetable oils. The catalysts were synthesized by the sequential incorporation of KF and/or Ca(NO3)2 solutions into the hydrogel upon microwave irradiation and then the as-obtained hydrogel was calcined at 800°C for 5 hours to eliminate the template and yield catalysts for the biodiesel productions. The prepared catalysts obtained by the different ways in the incorporation of ions into the hydrogel showed different physical properties and catalytic activities in the transesterification of soybean oil. All catalysts, except the low concentration of Ca(NO3)2, exhibiting the high activity yielding more than 90% FAME after 1 hour at 65°C, using oil to methanol molar ratio of 1:15 and 10 wt% of catalyst amounts.