Synthesis of regioisomerically pure triacylglycerols containing n‐3 very long‐chain polyunsaturated fatty acids

There is growing scientific evidence that consumption of n‐3 very long‐chain polyunsaturated fatty acids (n‐3 VLC‐PUFA) helps in brain and eye development, and protects against a range of common degenerative diseases. This has provided the impetus to the scientists to develop new and renewable sources for these important fatty acids so that the food industry is able to produce and market products fortified with n‐3 VLC‐PUFA. The bioactive efficacy and stability of food products containing n‐3 VLC‐PUFA may be determined not only by the amount of n‐3 VLC‐PUFA present but also by the positional distribution of these acids within the triacylglycerol (TAG) molecules (regiopurity). Studies of the effects of positional distribution on the functionality of n‐3 VLC‐PUFA containing oils have been hampered by a general lack of pure TAG regioisomers for experimentation. This paper reviews methods that have been used for the synthesis of TAG regioisomers containing n‐3 VLC‐PUFA, with special reference to those in which one n‐3 VLC‐PUFA occurs in combination with two long‐chain saturated acids.     

Mechanism Controlling High-Temperature Degradation of Sunflower Oil Triacylglycerols in the Absence of Oxygen

The aim of the present study is to describe the mechanism controlling heat-induced formation of sunflower oil triacylglycerol and fatty acid methyl ester oligomers. The unique combination of high-performance size-exclusion chromatography with hyphenated electrospray ionization mass spectrometry (MS), atmospheric pressure chemical ionization-MS, and high-temperature gas chromatography-MS techniques allows differentiating between radical coupling species and Diels–Alder cycloadducts. Targeted analysis of thermally degraded sunflower oils confirms the exact structures of various acyclic oligomers accompanied by less-abundant products of pericyclic transformations. A series of model experiments simulate the impact of dienophile nature on the course of Diels–Alder reactions. Thus, α-tocopherylquinone, δ-tocopherylquinone, and methyl-(E)-11-oxoundec-9-enoate are synthesized as naturally occurring dienophiles bearing electron-withdrawing groups. The geometry of poor dienophiles does not affect concerted cyclization, while the structure of electron deficient dienophiles can overcome low reactivity. Practical Application: In the absence of oxygen, heat-induced degradation of polyunsaturated triacylglycerols proceed predominantly via a radical pathway, whereas concerted reactions represent minor mechanisms. Sunflower oil triacylglycerol molecules in the system without propagation stage can be effectively protected by natural and/or synthetic antioxidants. Application of chelates is also recommended. However, antioxidant-derived quinones, such as α-tocopherylquinone, can enter the Diels–Alder reaction even more easily than dienophiles without electron-withdrawing groups. Unsaturated core aldehydes possess the same reactivity. Examination of the mechanism controlling high-temperature degradation of triacylglycerols is especially important for processing engineers in edible oil refineries and food technologists. New perspective may help them to minimize undesirable changes in polyunsaturated species.     

Triacylglycerols and Polar Lipids in Cow and Goat Milk are Differentially Affected by Various Lipid Supplemented Diets

This study characterizes milk triacylglycerol (TAG) and polar lipid (PL) fractions from cows and goats fed various lipid supplements modulating milk fat content. Twelve Holstein cows and 12 Alpine goats, at 86 ± 24.9 and 61 ± 1.8 days in milk, respectively, are allocated to one of 4 groups to receive diets supplemented with either corn oil [5% dry matter intake (DMI)] plus wheat starch (COS), marine algae powder (MAP; 1.5% DMI) or hydrogenated palm oil (HPO; 3% DMI), or a no-added-lipid control diet (CTL), according to a 4 × 4 Latin square design with 28 d experimental periods. Milk TAG and PL contents are determined by liquid chromatography-mass spectrometry (LC-MS). Multivariate analysis and ANOVA demonstrate major between-species differences in diet effects. In cows, COS specifically increases TAG 54:3 and 54:4 associated with milk fat depression (MFD), and increases the sum of phosphatidylcholines (PC) and phosphatidylinositols (PI). In addition to causing a MFD, MAP diet increases long-chain polyunsaturated TAG in both species, with higher magnitude in cows than in goats, and decreases the sum of PI in goats. HPO increases TAG 52:1 and the sum of PI in cows, but not in goats. Practical applications: Feed strategies can quickly and efficiently modulate the ruminant milk fat production and composition to improve nutritional quality for consumers. Certain starch-rich diets supplemented with polyunsaturated fatty acids (PUFA)-rich vegetable oils and diets supplemented with marine products (long-chain PUFA) reduce milk fat secretion and modify the milk fatty acid (FA) profile in cows, but not—or less so—in goats. Advanced analysis of both the TAG and PL fractions of milk fat is required to unravel these differences in lipid metabolism between cows and goats fed various lipid-supplemented diets. This study brings new insight on using nutritional strategies to control milk lipid composition according to ruminant species.     

Quantitative determination of triacylglycerol profile of structured lipid by capillary supercritical fluid chromatography and high-temperature gas chromatography

Two analytical methods have been developed for the qualitative and quantitative analyses of triacylglycerol profile of structured lipid (SL)-containing medium-chain and long-chain fatty acids. Supercritical fluid chromatography (SFC) was used in the first method. The SL was dissolved in chloroform/methanol, 95:5 (vol/vol), and analyzed directly using a super-critical fluid chromatograph equipped with temperature and density programming capabilities. No derivatization was required for sample preparation. An SB-methyl-100 capillary column (10 m, 100 μ i.d., 0.25 μ film thickness) was used for the separation of the triacylglycerol species and a flame-ionization detector (FID) was used for the detection. Supercritical fluid carbon dioxide was used as the mobile phase. In the second method, the SL was hydrogenated to complete saturation prior to analysis using gas chromatography at high temperatures of up to 375°C. A DB-5HT capillary column (30 m × 0.32 mm i.d., 0.1 μ film thickness) was used for the separation. FID was used for the detection and helium gas was used as mobile phase. The triacylglycerol species were separated and identified based on their equivalent carbon number (ECN), the total carbon number of the acyl side chains. A calibration curve was constructed using a triacylglycerol mixture containing known amounts of monoacyltriacylglycerol standard materials ranging from ECN 18 (trihexanoin) to ECN 66 (tridocosanoin). The novel triacylglycerol species, ECN 32–43, created by the interesterification of medium-chain triacylglycerol (MCT) and long-chain triacylglycerol (LCT) were separated and identified based on their retention times. These triacylglycerols, ECN 32–43, were absent in the physical mixture of MCT and LCT. The unique triacylglycerol specieis, ECN 32-43, were therefore selected as the fingerprinting region for the qualitative identification of the SL. Quantitation of the novel triacylglycerol species in the SL was achieved by using the integrated peak area of the new species. Both methods were employed successfully to distinguish the physical mixture from the corresponding interesterified SL. Results generated by the two methods were compared and found to be in good agreement.     

A rapid method for enzymatic synthesis and purification of the structured triacylglycerol, 1,3-dilauroyl-2-oleoyl-glycerol

A rapid method for synthesis and purification of the structured triacylglycerol (TAG), 1,3-dilauroyl-2-oleoylglycerol (LaOLa), has been developed. A fraction containing 70% LaOLa was obtained by enzymatic transesterification between triolein and lauric acid using Lipozyme IM, which has 1,3-regioselectivity on the glycerol moiety of TAG. The fraction was passed through a Bond Elut SI Column to remove mono-and diacylglycerols produced during the transesterification. The TAG fraction thus obtained was applied to a reversed-phase column, eluted with acetonitrile/tetrahydrofuran (8:2, vol/vol) to separate different TAG species. The LaOLa fraction obtained by this method was of greater than 99% purity. The concentrations of total fatty acids and fatty acids bound to the sn-2 position of this LaOLa fraction were determined by gas-liquid chromatography after hydrolysis by pancreatic lipase, to confirm the purity of stereospecific isomers in this fraction. The final purity of LaOLa was found to be greater than 95%, which was in good agreement with the result obtained by high-performance liquid chromatography using a Lichrosorb Si60 argentated column.     

A trimer plus a dimer-gallate reproduce the bioactivity described for an extract of grape seed procyanidins

The relationship between grape seed-derived procyanidin extract components and their bioactivity was explored. The monomeric and dimeric structures only acted as anti-inflammatory agents. Similarly, pure C1 trimer was highly effective on LPS-activated macrophages. To reproduce all of the bioactivities of the total extract, a fraction enriched with trimeric structures was needed. This trimeric-enriched fraction was divided into subfractions, the most bioactive of which contained two compounds with a molecular weight equal to a trimer (865) and a dimer-gallate (729), according to spectrometric analysis. Thus, it may be concluded that a mixture of both molecules reproduces the bioactivity in glucose metabolism (3T3-L1), lipid metabolism (HepG2) and macrophage functionality (RAW 264.6).     

Effects of dietary flaxseed oil on cholesterol metabolism of hamsters

High-fat/cholesterol diets (HFCD) formulated by addition of butter (BU), coconut oil (CO), or flaxseed oil (FX) enhanced (P < 0.05) serum lipids of hamsters compared to the low-fat/cholesterol diet (Control). However, FX groups showed a hypocholesterolaemic effect compared to CO and BU groups. Lower (P < 0.05) hepatic triacylglycerol and cholesterol contents were measured in FX groups than those of CO and BU groups; whereas, higher (P < 0.05) faecal triacylglycerol and cholesterol contents were observed in FX groups. HMG-CoA reductase mRNA expression was upregulated (P < 0.05) by HFCD, whilst FX groups showed no (P > 0.05) influence on LDL-receptor mRNA expression compared to that of Control groups; however, higher (P < 0.05) than those of CO and BU groups. Meanwhile, there was a tendency towards higher CYP7A1 expression in the CO or FX group than the BU group. Thus, the hypocholesterolaemic effect of FX might result from increases of LDL-receptor mRNA expression, and cholesterol catabolism/output.     

Chemical synthesis of tricaproin, trienantin and tricaprylin

In the present work, we optimised the glycerol + caproic acid/enantic acid/caprylic acid esterification conditions, using chemical routes, in order to get tricaproin, trienantin and tricaprylin in a medium solely composed of reagents. The option for cleaner conditions, without catalyst and solvent was preferred. The best conditions for tricaprylin and trienantin chemical synthesis were 6 h of contact, with a temperature around 160 ± 5 °C, followed by a 20 h of contact using higher temperatures (around 200 ± 5 °C). The volatility of caproic acid impeded the prolonged use of higher temperatures and this limited the yield of tricaproin that could be obtained. The synthesis of these triacylglycerols in the absence of both solvent and catalyst is of importance in consideration of the possible nutritional use of the products so formed.     

Two Triacylglycerol Lipases Are Negative Regulators of Chilling Stress Tolerance in Arabidopsis

Cold stress is one of the abiotic stress conditions that severely limit plant growth and development and productivity. Triacylglycerol lipases are important metabolic enzymes for the catabolism of triacylglycerols and, therefore, play important roles in cellular activities including seed germination and early seedling establishment. However, whether they play a role in cold stress responses remains unknown. In this study, we characterized two Arabidopsis triacylglycerol lipases, MPL1 and LIP1 and defined their role in cold stress. The expression of MPL1 and LIP1 is reduced by cold stress, suggesting that they may be negative factors related to cold stress. Indeed, we found that loss-of-function of MPL1 and LIP1 resulted in increased cold tolerance and that the mpl1lip1 double mutant displayed an additive effect on cold tolerance. We performed RNA-seq analysis to reveal the global effect of the mpl1 and lip1 mutations on gene expression under cold stress. The mpl1 mutation had a small effect on gene expression under both under control and cold stress conditions whereas the lip1 mutation caused a much stronger effect on gene expression under control and cold stress conditions. The mpl1lip1 double mutant had a moderate effect on gene expression under control and cold stress conditions. Together, our results indicate that MPL1 and LIP1 triacylglycerol lipases are negative regulators of cold tolerance without any side effects on growth in Arabidopsis and that they might be ideal candidates for breeding cold-tolerant crops through genome editing technology.     

甘三酯指纹图谱相似度在芝麻油掺混检测中的应用

探索了HPLC-ELSD测定油脂甘三酯的方法,并测定了纯芝麻油与不同掺入比例的大豆油-芝麻油,葵花籽油-芝麻油的甘三酯指纹图谱,建立了纯芝麻油的甘三酯指纹图谱数据库。通过计算掺混油脂和纯芝麻油数据库的指纹图谱相似度,对掺混油脂进行检验。当大豆、葵花籽油的掺入量达到3%时,即可做出是否为掺混油脂的准确判定。