Mixture Of Free Fatty Acids Research Articles

Development of a quantitative GC-FID method for the determination of stearoyl-lactylates (E481/482) in foods

Sodium and calcium salts of stearoyl-lactylates (SLs) are food emulsifiers especially used in bread and bakery products to improve texture. They should be used at the lowest level at which the desired technological effect is achieved in a specific food category and at amounts not exceeding the maximums set by European Commission Regulation No. 1129/2011. In order to be able to evaluate whether these emulsifiers are used correctly but also to evaluate whether the commercial additive formulations comply with legislation, a quantitative GC-FID method was developed. An internal standard (nonadecanoyl-1-lactylate) was synthesized in-house and pure ester standards were isolated from commercial additive formulations. The method showed a limit of detection of 0.04 and a limit of quantification of 0.12 mg esters ml−1. The commercial additive formulations analysed proved to be complex mixtures of free lactic and fatty acids together with only 50–60% esters. Besides SLs important amounts of palmitoyl-lactylates were present. Different food matrices (with low- and high-fat contents) were spiked with commercial SL formulations and recoveries ranged between 85% and 109%. Determination of SLs in commercial foods (such as bakery and bread) indicated that pre-treatment with amylase was essential to determine accurately the SL content due to the interaction of SL with the amylose.

Liraglutide reduces lipid accumulation in steatotic L-02 cells by enhancing autophagy

Simple hepatic steatosis is the early stage of non-alcoholic fatty liver disease and is recognized as a benign process. Previous studies show that glucagon-like peptide-1 has great potential in improving hepatic steatosis. Recent data have revealed that inhibiting autophagy exacerbates lipid accumulation in hepatocytes. Therefore, the present study aimed to determine the effects of liraglutide (LG) on simple hepatic steatosis and the possible role of autophagy. Firstly, steatotic L-02 cells were induced by incubating L-02 cells with 1 mmol/l free fatty acid (FFA) mixture (oleic acid and palmitic acid at a molar ratio of 2:1) for 24 h. Intracellular lipid accumulation, cell viability, oxidative stress and apoptosis were evaluated. Secondly, steatotic L-02 cells were treated with 10 or 100 nmol/l LG, 100 nmol/l LG plus 3-methyladenine (3-MA), or rapamycin for 24 h, and then lipid accumulation was measured. Next, the degree of lipid accumulation and the intensity of autophagy were assessed. Oil red O staining and triglyceride quantification demonstrated notable steatosis in L-02 cells following exposure to 1 mmol/l FFA mixture for 24 h. There was no significant cytotoxicity, oxidative stress or apoptosis in steatotic L-02 cells. Treatment with 100 nmol/l LG reduced lipid accumulation in steatotic L-02 cells and increased the mRNA levels of microtubule-associated protein 1 light chain 3B. Additionally, it enhanced the autophagic flux in steatotic L-02 cells, as measured by western blot analysis and shown by electron microscopy. Additionally, 3-MA weakened the ability of LG to improve hepatic steatosis and enhance autophagy. Our data indicate that LG reduces the lipid accumulation in steatotic L-02 cells, and the activation of autophagy plays a significant role in this process.

One-week high-fat diet leads to reduced toll-like receptor 2 expression and function in young healthy men

Toll-like receptor 2 (TLR2) is implicated in inflammatory responses to high-fat diet (HFD)–induced obesity in rodents, but human HFD studies examining TLR2-mediated immune responses are lacking. Our aim was to determine whether HFD affected TLR2 function in humans. We hypothesized that a short-term HFD in humans would impair TLR2-mediated immune function. Fasting blood samples were obtained from healthy young men (N = 9) before and after a 7-day HFD. Toll-like receptor 2 function was assessed in ex vivo whole blood cultures stimulated with the TLR2 agonist N-palmitoyl-S-[2,3-bis[palmitoyloxy]-[2RS]-propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine (Pam3-Cys-SK4). Peripheral blood mononuclear cells (PBMCs) were isolated to examine TLR2, TLR4, and p47 subunit of nicotinamide adenine dinucleotide phosphate oxidase (p47phox) protein expression via Western blotting. Pam3-Cys-SK4–stimulated secretion of interleukin-1β (−35%, P = .005), interleukin-6 (−32%, P = .01), and tumor necrosis factor–α (−33%, P = .06) was reduced following the HFD. High-fat diet resulted in decreased TLR2 (P = .049) and p47phox (P = .037) protein expression from PBMCs. To mimic lipid overload ex vivo, follow-up experiments were performed in whole blood cultures exposed to a mixture of free fatty acids for 24 hours; and surface protein expression of TLR2 and TLR4 on CD14+ monocytes was measured by flow cytometry. Free fatty acid exposure for 24 hours ex vivo reduced monocyte TLR2 levels by about 20% (P = .028). A 7-day HFD in young healthy men resulted in impaired TLR2 function. Decreased TLR2 and p47phox protein expression in PBMCs, possibly due to excess free fatty acids, may mediate this response. Our current findings indicate that impaired TLR2 response after HFD might be partially responsible for increased risk of infection in diet-induced obesity.

Cholesterol efflux from THP-1 macrophages is impaired by the fatty acid component from lipoprotein hydrolysis by lipoprotein lipase

Lipoprotein lipase (LPL) is an extracellular lipase that primarily hydrolyzes triglycerides within circulating lipoproteins. Macrophage LPL contributes to atherogenesis, but the mechanisms behind it are poorly understood. We hypothesized that the products of lipoprotein hydrolysis generated by LPL promote atherogenesis by inhibiting the cholesterol efflux ability by macrophages. To test this hypothesis, we treated human THP-1 macrophages with total lipoproteins that were hydrolyzed by LPL and we found significantly reduced transcript levels for the cholesterol transporters ATP binding cassette transporter A1 (ABCA1), ABCG1, and scavenger receptor BI. These decreases were likely due to significant reductions for the nuclear receptors liver-X-receptor-α, peroxisome proliferator activated receptor (PPAR)-α, and PPAR-γ. We prepared a mixture of free fatty acids (FFA) that represented the ratios of FFA species within lipoprotein hydrolysis products, and we found that the FFA mixture also significantly reduced cholesterol transporters and nuclear receptors. Finally, we tested the efflux of cholesterol from THP-1 macrophages to apolipoprotein A-I, and we found that the treatment of THP-1 macrophages with the FFA mixture significantly attenuated cholesterol efflux. Overall, these data show that the FFA component of lipoprotein hydrolysis products generated by LPL may promote atherogenesis by inhibiting cholesterol efflux, which partially explains the pro-atherogenic role of macrophage LPL.

Anti-lipoapoptotic effect of Artemisia capillaris extract on free fatty acids-induced HepG2 cells.

BackgroundArtemisia capillaris (AC) has been recognized as one of the promising candidates for hepatoprotective, hypoglycemic, hypolipidemic, antiobesitic and anti-inflammatory therapeutic effectiveness. This study evaluated the inherent mechanism and anti-apoptotic activity of 30% ethanol extract of AC (AC extract) 100 μg/ml on free fatty acids (FFAs)-induced HepG2 cellular steatosis and lipoapoptosis.MethodsHepatic steatosis was induced by culturing HepG2 cells with a FFAs mixture (oleic and palmitic acid at the proportion of 2:1) for 24 h, thus ultimately giving rise to lipoapoptosis. Cell viability and lipid accumulation were detected by MTT assay and Oil Red O staining method respectively and Caspase-3, −9, Bax, Bcl-2, p-JNK and PUMA were measured for lipoapoptosis after 24 hours.ResultsAC extract significantly improved the FFAs-induced steatosis without cytotoxicity and Caspase-3, −9, Bax and Bcl-2 were modulated profitably to HepG2 cells after AC treatment. In addition, AC extract inhibited the activation of c-Jun NH2 terminal kinase (JNK) and PUMA, which mechanism is related to non-alcoholic steatohepatitis (NASH).ConclusionsCombined together, AC extract exerted an obvious hypolipidemic and anti-apoptotic effect, indicating that AC extract might have potential therapeutic herb against NASH.

Novel Biobased Nonionic Surfactants: Synthesis, Surface Activity and Corrosion Inhibition Efficiency Against Aluminum Alloy Dissolution in Acidic Media

AbstractFour nonionic surfactants were prepared from the reaction of propylene oxide with oleic acid, linoleic acid, and the free fatty acid mixture from hydrolysis of jatropha oil. The chemical structures of the prepared surfactants were confirmed using IR and NMR spectroscopy. The surface activities of the prepared surfactants were dependent on the polypropylene oxide chain length and also on the nature of the alkyl chains. The nonionic surfactants were evaluated at different concentrations as corrosion inhibitors against the corrosion of Al 6061 aluminum in 2 M HCl solution. The corrosion inhibition tendencies of the surfactants were completely dependent on the fatty acid ratio in the jatropha oil and also on the polypropylene oxide chain length. The corrosion inhibition efficiencies of the surfactants were correlated to their chemical structure and their surface activities.

The effect of the chain length distribution of free fatty acids on the mixing properties of stratum corneum model membranes

The stratum corneum (SC) plays a fundamental role in the barrier function of the skin. The SC consists of corneocytes embedded in a lipid matrix. The main lipid classes in the lipid matrix are ceramides (CERs), cholesterol (CHOL) and free fatty acids (FFAs). The aim of this study was to examine the effect of the chain length of FFAs on the thermotropic phase behavior and mixing properties of SC lipids. Fourier transform infrared spectroscopy and Raman imaging spectroscopy were used to study the mixing properties using either protonated or deuterated FFAs. We selected SC model lipid mixtures containing only a single CER, CHOL and either a single FFA or a mixture of FFAs mimicking the FFA SC composition. The single CER consists of a sphingoid base with 18 carbon atoms and an acyl chain with a chain length of 24 carbon atoms. When using lignoceric acid (24 carbon atoms) or a mixture of FFAs, the CER and FFAs participated in mixed crystals, but hydration of the mixtures induced a slight phase separation between CER and FFA. The mixed crystalline structures did not phase separate during storage even up to a time period of 3months. When using palmitic acid (16 carbon atoms), a slight phase separation was observed between FFA and CER. This phase separation was clearly enhanced during hydration and storage. In conclusion, the thermotropic phase behavior and the mixing properties of the SC lipid mixtures were shown to strongly depend on the chain length and chain length distribution of FFAs, while hydration enhanced the phase separation.

Features of the enzymatic hydrolysis of castor oil

Ricinoleic acid, which is used in medicine and veterinary science and is the initial material in the organic synthesis of various valuable products, is obtained via the hydrolysis of castor oil. The enzymatic hydrolysis of castor oil, which allows us to conduct the process under mild conditions (in the temperature range of 35–45°C and without high pressures) is a promising method for obtaining ricinoleic acid. This work demonstrates the feasibility of the enzymatic hydrolysis of castor oil with lipase from Candida rugosa in oil-water systems without an emulsifier. We propose a method for hydrolysis without emulsifiers, simplifying the process of isolating the target product (a mixture of free fatty acids in which ricinoleic acid predominates) and thus the relevant technology. The catalyst that we use ensures the environmental friendliness of the process. Our selection of the experimental conditions for hydrolysis resulted in a 47% yield of fatty acids.

Chemoenzymatic Method for Producing Stearidonic Acid Concentrates from Stearidonic Acid Soybean Oil

AbstractThe aim of this study was to produce stearidonic acid (SDA, 18:4n‐3) omega‐3 concentrates from 25 % SDA soybean oil (SDASO) by enzymatic acidolysis. Substrates were prepared by chemical and enzymatic hydrolysis of SDASO. A 62 % SDA free fatty acid mixture (SDA‐FFA) was obtained by low temperature crystallization of the chemical hydrolyzate while partial hydrolysis of SDASO by non‐immobilized lipase AY 30 (Candida rugosa) yielded a 51 % SDA acylglycerol mixture (SDA‐GLY). Reaction conditions for acidolysis between SDA‐FFA and SDA‐GLY were optimized using response surface methodology (RSM). Incorporation of SDA into SDA‐GLY by immobilized Lipozyme RM IM (Rhizomucor miehei) and non‐immobilized Lipomod 34P‐LO34P (C. cylindracea [rugosa]) lipases were modeled under varying levels of the substrate molar ratio (Sr), temperature (T), and time (t). Optimal conditions for production of a 60 % SDA concentrate were predicted to be Sr = 4.8, T = 65 °C and t = 8 h for Lipozyme RM IM and Sr = 5.0, T = 43 °C and t = 48 h for Lipomod 34P‐L034P. The model was verified experimentally by gram scale synthesis under optimal conditions which resulted in the production of 59.98 and 58.98 % SDA concentrates (≥96 % triacylglycerols) by Lipozyme RM IM and Lipomod 34P‐L034P, respectively.

Differently Saturated Fatty Acids Can Be Differentiated by 31P NMR Subsequent to Derivatization with 2-Chloro-4,4,5,5-tetramethyldioxaphospholane: A Cautionary Note

The analysis of free fatty acid (FFA) mixtures is a very important but, even nowadays, challenging task. This particularly applies as the so far most commonly used technique-gas chromatography/mass spectrometry (GC/MS)-is tedious and time-consuming. It has been convincingly shown ( Spyros, A.; Dais, P. J. Agric. Food Chem. 2000, 48, 802 - 5) that FFA may be analyzed by (31)P NMR subsequent to derivatization with 2-chloro-4,4,5,5-tetramethyldioxaphospholane (CTDP). However, it was also indicated that differently unsaturated FFAs result in the same (31)P NMR chemical shift and cannot be differentiated. Therefore, only the overall fatty acid content of a sample can be determined by the CTDP assay. In contrast, we will show here by using high-field NMR (600 MHz spectrometer, i.e., 242.884 MHz for (31)P) that the CTDP assay may be used to differentiate FFAs that have pronounced differences in their double bond contents: saturated fatty acids (16:0), moderately unsaturated (18:1, 18:2), highly unsaturated (20:4), and extremely unsaturated fatty acids (22:6) result in slightly different chemical shifts. The same applies for oxidized fatty acids. Finally, it will also be shown that the CTDP derivatization products decompose in a time-dependent manner. Therefore, all investigations must adhere to a strict time regime.

Separation of cis-Fatty Acids from Saturated and trans-Fatty Acids by Nanoporous Polydicyclopentadiene Membranes

This article describes the separation of mixtures of fatty acid salts using a new organic solvent nanofiltration membrane based on polydicyclopentadiene (PDCPD). Mixtures of free fatty acids could not be separated by the membranes because they permeated at similar rates. When triisobutylamine was added to the fatty acids, the cis-fatty acid salts (oleic, petroselinic, vaccenic, linoleic, and linolenic acid) had slower permeation though the membranes than saturated (stearic acid) and trans-fatty acid (elaidic acid) salts. The reason for the difference in permeation was due to the formation of stable salt pairs between the amine and fatty acids that increased their cross-sectional areas. The fatty acid salts derived from saturated and trans-fatty acids were smaller than the critical area cutoff for the PDCPD membranes, so they readily permeated. In contrast, the fatty acid salts derived from the cis-fatty acids had critical areas larger than critical area cutoff of the PDPCD membranes and had slowed permeation. The partitioning coefficients of fatty acids and fatty acid salts were investigated to demonstrate that they were not responsible for the difference in permeation. The use of pressure was investigated to greatly accelerate the permeation through the membranes. For a solvent mixture of 35/65 (v/v) toluene/hexanes, the permeation of solvent was approximately 39 L m(-2) h(-1). This value is similar to values reported for permeation through membranes used in industry. The separation of a mixture of fatty acids based on the composition of soybean oil was investigated using pressure. The saturated fatty acid salts were almost completely removed from the cis-fatty acid salts when iBu(3)N was used as the amine to form the salt pairs. The separation of the cis-fatty acids found in soybean oil was investigated with Pr(3)N as the amine. The oleic acid salt (oleic acid has one cis double bond) preferentially permeated the membrane while the linoleic (two cis double bonds) and linolenic (three cis double bonds) salts were partly retained. The separation of fatty acids using membranes may have real applications in industry to purify fatty acids on a large scale.

Novel homodimer model of the β-adrenergic receptor in complex with free fatty acids and cholesterol: first-principles calculation studies.

We propose a theoretical novel homodimer model of the β- adrenergic receptor (βAR) in complex with a heterogeneous mixture of free fatty acids (FFAs) and cholesterol based on first-principles calculations. We used the density-functional-based tight binding with dispersion (DFTB-D) method, which accurately evaluates van der Waals interactions between FFAs and amino acid residues in the receptor. The calculations suggest that a stable homodimer of bAR can form a complex with FFAs and cholesterol by extensive van der Waals interactions in the cell membrane, and that the heterogeneous composition of the FFAs is important for the stability of the homodimer complex. The stable van der Waals interactions propagate from one of the bAR to the other through the cholesterol and FFAs in the homodimer complex. The energy propagation in the complex has the potential to enhance molecular signaling in adipocytes, because the stability of the complex can influence anti-adiposity effects after oral treatment of the FFA components.

Simultaneous conversion of triacylglycerides and fatty acids into fatty acid methyl esters using organometallic tin(IV) compounds as catalysts

Three Sn(IV) complexes, named butyl stannoic acid (BTA), di-n-butyl-oxo-stannane (DBTO) and dibutyltin dilaurate (DBTDL), were initially tested as catalysts for esterification reaction of fatty acids in the presence of methanol as alcoholysis agent. Parameters like reaction time, temperature, and catalyst amount were systematically evaluated in this work. All complexes were active at relative high reaction temperatures, but BTA displayed the highest activity. Former studies have already demonstrated that these same complexes display good catalytic activity in methanolysis of triacylglycerides (TAGs). These results prompted us to test BTA catalyst also in simultaneous esterification/transesterification reactions from a mixture of free fatty acids (FFAs) and TAGs in the presence of methanol. BTA was able to convert the mixtures in to fatty acid methyl esters (FAMEs) with good yields, with simple isolation processes. The results obtained and discussed in this work can help the development of new catalytic systems to biodiesel production from oils with very high acid content.

Oxidative stress impairs HIF1α activation: a novel mechanism for increased vulnerability of steatotic hepatocytes to hypoxic stress

Steatosis increases the sensitivity of hepatocytes to hypoxic injury. Thus, this study was designed to elucidate the role of hypoxia-inducible factor-1α (HIF1α) in steatotic hepatocytes during hypoxia. AML12 hepatocytes and isolated rat hepatocytes were treated with a free fatty acid mixture of oleate and palmitate (2:1, 1mM) for 18h, which generated intrahepatocyte fat accumulation. The cells were then exposed to hypoxia (1% oxygen, 6–24h). After hypoxia, a further increase in cellular fat accumulation was seen. In steatotic hepatocytes, a decreased HIF1α activation by hypoxia was observed. The capacity of these cells to express HIF1α-dependent genes responsible for the utilization of nutrients for energy was also impaired. This resulted in significantly lower intracellular ATP levels and greater cell death in steatotic hepatocytes compared with control hepatocytes. In contrast, overexpression of constitutively active HIF1α significantly increased cell viability as well as ATP and GLUT1 mRNA levels in steatotic hepatocytes under hypoxia. Hypoxia significantly enhanced HIF1α mRNA levels in control but not in steatotic hepatocytes. Concomitantly, an increase in oxidative stress was found in steatotic hepatocytes under hypoxic conditions compared with control cells. This included higher reactive oxygen species generation, lower cellular and nuclear GSH levels, and higher accumulation of 4-hydroxynonenal protein adducts. Hypoxia-mediated oxidative stress was accompanied by inactivation of basal nuclear factor-κB (NF-κB) DNA binding. Treatment with N-acetyl-l-cysteine, a reducing agent, improved NF-κB DNA-binding capacity and restored HIF1α induction. Conversely, overexpression of an NF-κB super-suppressor in control hepatocytes (IκBαΔN-transfected cells) resulted in complete inhibition of HIF1α expression, confirming that indeed NF-κB regulates HIF1α expression in hypoxic hepatocytes. In conclusion, hypoxia in combination with hepatic steatosis was shown to promote augmented oxidative stress, leading to NF-κB inactivation and impaired HIF1α induction and thereby increased susceptibility to hypoxic injury.

Dynamic monitoring of β-cell injury with impedance and rescue by glucagon-like peptide-1

β-Cell injury plays an important role in the development of type 1 and type 2 diabetes. Most of the β-cell bioassays depend on labeling or endpoint assessments that might not capture the true physiology or pathology of the injury process. In the current study, we dynamically detected a broad range of pathological and pharmacological responses to four toxicants (cytokine mixture, free fatty acid mixture, streptozotocin, and hydrogen peroxide) in living β-cells (INS-1E and MIN6) by a label-free, cell-based assay system named xCELLigence, codeveloped by ACEA Biosciences and Roche Diagnostics. Our results suggest that the impedance readout is highly sensitive and provides more information than some of the conventional endpoint cytotoxicity assays for β-cell injury such as the Cell Counting Kit-8 (CCK-8) and morphology measurements. Furthermore, this system was used to evaluate the anti-injury effects of glucagon-like peptide-1 (GLP-1) and its nonpeptidic mimetic Boc5 by monitoring responses to four toxicants in two β-cell lines. This study confirms that the protective property of Boc5 on β-cells is similar to that of GLP-1.

Catalytic Transformation of Tall Oil into Biocomponent of Diesel Fuel

One of the conventional kraft pulp mills produce crude tall oil which is a mixture of free fatty acids, resin acids, sterols, terpenoid compounds, and many others. This study is devoted to the issue of direct transformation of crude tall oil in a mixture with straight-run atmospheric gas oil to liquid fuels using three different commercial hydrotreating catalysts. Diesel fuel production is an alternative to incineration of these materials. High catalytic activity was achieved for all tested catalysts in temperature range 360–380°C, under 5.5 MPa hydrogen pressure and ratio H2/feedstock 500–1000 l/l. Crude tall oil can be converted to diesel oil component via simultaneous refining with straight-run atmospheric gas oil on NiMo/Al2O3and NiW/Al2O3-zeolite catalysts. All tested catalysts had very good hydrodenitrogenation activity and high liquid yield were at tested conditions.

Optimal Design of Biodiesel Production Process from Waste Cooking Palm Oil

Abstract A design methodology for biodiesel production from waste cooking palm oil is proposed. The proposed method is flexible to the biodiesel process using various catalyst types: alkali and acid catalyst in homogenous and heterogeneous forms, and different process: enzyme process and supercritical process. A two-step approach of hydrolysis and esterification processes is also considered. Waste cooking palm oil consists of a mixture of triglyceride (e.g., trilaurin, tripalmitin, triolein, tristearin, trilinolein and trilinolenin) and free fatty acids (e.g., lauric acid, palmitic acid, stearic acid, oleic acid, linoleic and linolenic acid). A driving force approach and thermodynamic insight are employed to design separation units (e.g., flash separator and distillation) minimizing the energy consumption. Steady-state simulations of the developed biodiesel processes are performed and economic analysis is used to find a suitable biodiesel process. The results show that based on a net present value, the heterogeneous acid catalyzed process is the best process for biodiesel production. With the design methodology, the proposed biodiesel process can save the energy requirement of 41.5%, compared with a conventional process.

Inhibition of fetal bone development through epigenetic down‐regulation of HoxA10 in obese rats fed high‐fat diet

Epidemiological studies show that maternal obesity during intrauterine and early postnatal life increases the risk of low bone mass and fracture later in life. Here, we show that bone development is inhibited in gestational embryonic day 18.5 (E18.5) embryos from rat dams made obese by feeding a high-fat diet (HFD). Moreover, fetal rat osteogenic calvarial cells (FOCCs) from these obese dams have significantly less potential to develop into mature osteoblasts compared to cells from AIN-93G diet-fed controls. Profiling of transcriptional genes for osteogenesis revealed a profound decrease in the homeodomain-containing factor A10 (HoxA10) in FOCCs from fetuses of HFD-induced obese dams. Significant methylation of the HoxA10 promoter was found in those FOCCs, as well as in mouse ST2 cells treated with a mixture of free fatty acids similar to that found in serum from HFD-induced obese rats. This was accompanied by lower expression of osteogenic markers, but higher levels of PPARγ. Control FOCCs depleted of the HoxA10 gene (shRNA) ex vivo behave similarly to cells from fetuses of obese dams; conversely, overexpression of HoxA10 gene in FOCCs from HFD rats exhibit the same phenotype as controls. Treatment of FOCCs from control rats or of ST2 cells with an artificial mixture of free fatty acids significantly down-regulated HoxA10 protein expression, and cells exhibited adipocyte-like properties. These results suggest that maternal obesity impairs fetal skeletal development through down-regulation of the HoxA10 gene, which may lead to an increase in the prevalence of low bone mass in the offspring later in life.

Enrichment of palm olein with long chain polyunsaturated fatty acids by enzymatic acidolysis

Long-chain polyunsaturated fatty acids (LCPUFA) such as docosahexaenoic (DHA) and arachidonic (ARA) acids have great benefits for the development and maintenance of human brain functions. The production of structured lipid (SL) by acidolysis of palm olein with a free fatty acid mixture obtained from DHASCO® and ARASCO®, catalyzed by Novozym 435® in hexane was optimized by response surface methodology (RSM). Three independent factors chosen were substrate mole ratio of total free fatty acids to palm olein (6–18 mol/mol), reaction temperature (55–65 °C), and reaction time (12–24 h). Good quadratic models were obtained for the incorporation of DHA + ARA (response 1) and their incorporation at the sn-2 position (response 2) by multiple regression and backward elimination. The models were verified by carrying out acidolysis reaction at several random combinations of the independent factors. The optimal condition generated from the models resulted in 25.25 g/100 g total incorporation of DHA + ARA and 17.20 g/100 g DHA + ARA incorporation at the sn-2 position. The fatty acid composition of SL was comparable to that of human milk fat. The SL produced in this study has potential for use in infant formulas as well as in nutraceutical applications for pregnant women.

Impact of the carotenoid astaxanthin on phagocytic capacity and ROS/RNS production of human neutrophils treated with free fatty acids and high glucose

BackgroundThe purpose of the present study was to evaluate the effect of carotenoid astaxanthin (ASTA) on human neutrophils treated with high glucose and free fatty acids (FFA) on the phagocytic capacity and reactive oxygen/nitrogen species production. MethodsThe following parameters were evaluated: phagocytic capacity of neutrophils by using zymosan particles, intracellular and extracellular superoxide anion (lucigenin and DHE probes), hydrogen peroxide (H2O2 — phenol red), nitric oxide (Griess reagent) production, and maximal activity of G6PDH. ResultsThere was a decreased phagocytic capacity of human neutrophils treated with high glucose (30mM) or FFA (0.1mM) and a partial restoring of the phagocytic capacity after ASTA-treatment was observed. ROS and RNS production was increased in neutrophils due to both high glucose and FFA. This increase in ROS/RNS production was also partially prevented by ASTA treatment. Both glucose and FFA increased the G6PDH activity. We show that ASTA provides a modest improvement of cellular functions after cells have been treated with high glucose and FFA. ConclusionsIn summary, this study showed that both high glucose and a mixture of FFA are potent inducers of ROS/RNS production on neutrophils as observed by higher levels of superoxide anion, hydrogen peroxide and NO production. Also, these metabolites decrease the phagocytic capacity of neutrophils and increase the G6PDH activity. Overall, ASTA-treatment was able to reduce partially ROS/RNS production by reducing the availability of NADPH and recover phagocytic capacity of neutrophils.