Molecular weight distribution and regioisomeric structure of triacylglycerols in some common human milk substitutes

Triacylglycerol (TAG) molecular weight distribution and regioisomeric structure of selected molecular weight species in human milk and in 32 human milk substitutes was determined. Negative ion chemical ionization mass spectrometry was used to determine the molecular weight distribution and collisionally induced dissociation tandem mass spectrometry applied to identify the sn-2 and sn-1/3 positions of fatty acids in TAG. The main molecular weight species of human milk TAG in decreasing order of abundance were 52∶2, 52∶3, 52∶1, 54∶3, 50∶2, 50∶1, 54∶4, 48∶1, 54∶2, 48∶2, 46∶1, 52∶4, and 50∶3 (acyl carbon number/number of double bonds), constituting 83 mol% of total TAG molecular species. In human milk substitutes, the proportion of the corresponding molecular weight species varied from 33 to 87 mol%. The main TAG regioisomers within the molecular weight species 52∶2, 52∶3, and 50∶1 in human milk were 18∶1-16∶0-18∶1 (83 mol%), 18∶1-16∶0-18∶2 (83 mol%), and 18∶1-16∶0-16∶0 (80 mol%), respectively. In human milk substitutes, the corresponding proportions varied in a wide range of 0–82 mol%, 0–100 mol%, and 0–73 mol%, respectively. Although TAG structures in some human milk substitutes closely resembled those in human milk, the great variation among samples leads to the conclusion that it is still possible to improve the TAG composition in human milk substitutes by applying novel methods to synthesize structured TAG.     

Positional distribution of fatty acids in the triacylglycerols of developing oil palm fruit

The pattern of accumulation of triacylglycerols, their fatty acid compositions and the positional distribution of the fatty acids at thesn-2- andsn-1,3-positions of the triacylglycerol molecules at progressive stages of oil palm fruit development were determined. There was an exponential rate of increase of triacylglycerols and their fatty acids toward the end of fruit development. The fatty acid composition of the triacylglycerols in the early stages of development, prior to active accumulation, was more or less similar, but differed appreciably from the later stages, and the transition of fatty acid composition toward that of normal palm oil occurred at around 16 wk after anthesis (WAA) and stabilized at 20 WAA. All fatty acids increased in terms of absolute quantity. There was an overall consistency in fatty acid positional distribution, irrespective of development stage. More saturated fatty acids were found to be esterified at thesn-1,3-positions and more unsaturated fatty acids at thesn-2-position of triacylglycerol. Higher rate of incorporation of 16:0 at the 1,3-positions during the active phase of triacylglycerol synthesis was observed, while 18:1 acid exhibited a reverse trend.     

Synthesis of triacylglycerol from polyunsaturated fatty acid by immobilized lipase

More than 95% of polyunsaturated acid (PUFA) was converted to triacylglycerol by immobilized lipase fromCandida antarctica orRhizomucor miehei. The esterification was carried out at 50–60°C with shaking and dehydration for 24 h. The substrates consisted of glycerol and free fatty acid or ethyl esters of the fatty acid at a 1∶3 molar ratio. The docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA) in the substrate polymerized during the reaction, and they required 5–10% more than the stoichiometric amount to compensate for the PUFA loss. On the contrary, ethyl esters of DHA and EPA were not polymerized. Pure tridocosahexaenoyl, trieicosapentaenoly and triarachidonoyl glycerol were isolated after passing the product through a basic aluminum oxide column. Industrial feasibility of this process was discussed for the ethyl ester as substrate. Portions of this article were presented at the annual meeting of the Japan Society for Bioscience, Biotechnology, and Agrochemistry and at the annual meeting of The Japan Oil Chemists' Society held in Kyoto, Japan, March 31, 1991, and in Hamamatsu, Japan, October 4, 1991, respectively.     

13C nuclear magnetic resonance spectroscopic analysis of the triacylglycerol composition ofBiota orientalis and carrot seed oil

¹³C nuclear magnetic resonance (NMR) spectroscopic analysis of the whole oil (triacylglycerols) ofBiota orientalis seeds confirms the presence of oleate [18:1(9Z)], linoleate [18:2(9Z, 12Z)], linolenate [18:3((9Z, 12Z, 15Z)], 20:3 (5Z, 11Z, 14Z), 20:4(5Z, 11Z, 14Z, 17Z), and saturated fatty acids in the acyl groups by comparing the observed carbon shifts with previously established shift data for model triacylglycerols. This technique shows that the saturated, 20:3 and 20:4 fatty acids are distributed mainly in the α-acyl positions, whereas oleate, linoleate, and linolenate are randomly acylated to the α- and β-positions of the glycerol “backbone”. Stereospecific hydrolysis of theBiota oil with pancreatic lipase, followed by chromatographic analysis of fatty esters, reveals the presence of trace amounts of 16:0(0.7%), 18:0(0.5%), 20:3 (0.4%), and 20:4 (1.3%) in the β-position of the glycerol “backbone”, which are undetectable by¹³C NMR technique on the whole oil. Semiquantitative assessment of the¹³C NMR signal intensities gives the relative percentages of the fatty acid distribution as: saturated 16:0, 18:0 (12.0% α-acyl), oleate (7.7% α-acyl 8.7% β-acyl), total linoleate and linolenate (31.7% α-acyl; 24.2% βacyl), total 20:3 and 20:4 (15.7% α-acyl). The¹³C NMR spectroscopic analysis of carrot seed oil identifies the presence of saturated (18:0), 18:1(6Z), 18:1(9Z), and 18:2(9Z, 12Z). The saturated fatty acid is found in the α-acyl positions. Semi-quantitative assessment of the signal intensities gives the relative percentages of the fatty acids as: 18:0 (4.5% α-acyl), 18:1(6Z) (49.6% α-acyl; 19.7% β-acyl), oleate (6.5% α-acyl; 8.6% β-acyl) and linoleate (5.2% α-acyl; 6.9% β-acyl).     

Mass spectrometric analysis of long-chain lipids

Electrospray and matrix assisted laser desorption ionization generate abundant molecular ion species from all known lipids that have long chain fatty acyl groups esterified or amidated to many different polar headgroup features. Molecular ion species include both positive ions from proton addition [M+H](+) and negative ions from proton abstraction [M-H](-) as well as positive ions from alkali metal attachment and negative ions from acetate or chloride attachment. Collisional activation of both MALDI and ESI behave very similarly in that generated molecular species yield product ions that reveal many structural features of the fatty acyl lipids that can be detected in tandem mass spectrometric experiments. For many lipid species, collision induced dissociation of the positive [M+H](+) reveals information about the polar headgroup, while collision induced dissociation of the negative [M-H](-) provides information about the fatty acyl chain. The mechanisms of formation of many of these lipid product ions have been studied in detail and many established pathways are reviewed here. Specific examples of mass spectrometric behavior of several molecular species are presented, including fatty acids, triacylglycerol, phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, ceramide, and sphingomeylin.     

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.