Acylglycerols Containing Trihydroxy Fatty Acids in Castor Oil and the Regiospecific Quantification of Triacylglycerols

Ricinoleate, a monohydroxy fatty acid in castor oil, has many industrial uses. Dihydroxy and trihydroxy fatty acids can also be used in industry. We report here the identification of diacylglycerols (DAG) and triacylglycerols (TAG) containing trihydroxy fatty acids in castor oil. The C₁₈ HPLC fractions of castor oil were used for mass spectrometry of the lithium adducts of acylglycerols to identify trihydroxy fatty acids and the acylglycerols containing trihydroxy fatty acids. Two DAG identified were triOH18:1–diOH18:1 and triOH18:0–OH18:1. Four TAG identified were triOH18:1–OH18:1–OH18:1, triOH18:0–OH18:1–OH18:1, triOH18:1–OH18:1–diOH18:1 and triOH18:0–OH18:1–diOH18:1. The structures of these two newly identified trihydroxy fatty acids were proposed as 11,12,13-trihydroxy-9-octadecenoic acid and 11,12,13-trihydroxyoctadecanoic acid. The locations of these trihydroxy fatty acids on the glycerol backbone were almost 100% at the sn-1,3 positions or at trace levels at the sn-2 position. The content of these acylglycerols containing trihydroxy fatty acids was at the level of about 1% or less in castor oil.     

Rapid Quantitative Determination of Squalene in Shark Liver Oils by Raman and IR Spectroscopy

Squalene is sourced predominantly from shark liver oils and to a lesser extent from plants such as olives. It is used for the production of surfactants, dyes, sunscreen, and cosmetics. The economic value of shark liver oil is directly related to the squalene content, which in turn is highly variable and species‐dependent. Presented here is a validated gas chromatography‐mass spectrometry analysis method for the quantitation of squalene in shark liver oils, with an accuracy of 99.0 %, precision of 0.23 % (standard deviation), and linearity of >0.999. The method has been used to measure the squalene concentration of 16 commercial shark liver oils. These reference squalene concentrations were related to infrared (IR) and Raman spectra of the same oils using partial least squares regression. The resultant models were suitable for the rapid quantitation of squalene in shark liver oils, with cross‐validation r² values of >0.98 and root mean square errors of validation of ≤4.3 % w/w. Independent test set validation of these models found mean absolute deviations of the 4.9 and 1.0 % w/w for the IR and Raman models, respectively. Both techniques were more accurate than results obtained by an industrial refractive index analysis method, which is used for rapid, cheap quantitation of squalene in shark liver oils. In particular, the Raman partial least squares regression was suited to quantitative squalene analysis. The intense and highly characteristic Raman bands of squalene made quantitative analysis possible irrespective of the lipid matrix.     

Vaccenic and Elaidic Acid Equally Esterify into Triacylglycerols,but Differently into Phospholipids of Fed Rat Liver Cells

Elaidic acid (trans-9-C_18:1 or trans-9) is assumed to exert atherogenic effects due to its double bond configuration. The possibility that trans-9 and vaccenic acid (trans-11-C_18:1 or trans-11), its positional isomer, were biochemically equivalent and interchangeable compounds, was investigated by reference to their cis-isomers through esterification-related activities using rat liver cells and subcellular fractions. In hepatocytes, both trans-C_18:1 were incorporated to the same extent in triacylglycerols, but trans-9 was more esterified than trans-11 into phospholipids (P < 0.05). Glycerol-3-phosphate acyltransferase activity in microsomes was lower with trans-11 than with trans-9, while this activity in mitochondria was ~40% greater with trans-11 than with trans-9 (P < 0.05). Activity of 2-lysophosphatidic acid acyltransferase in microsomes was of comparable extent with both trans isomers, but activity of 2-lysophosphatidylcholine acyltransferase was significantly greater with trans-9 than with trans-11 at P < 0.01. Lipoproteins secreted by hepatocytes reached equivalent levels in the presence of any isomers, but triacylglycerol production was more elevated with trans-11 than with trans-9 at P < 0.05. Cholesterol efflux from previously labelled hepatocytes was lower with trans-11 than with trans-9. When these cells were exposed to either trans-C_18:1, the gene expression of proteins involved in fatty acid esterification and lipoprotein synthesis was unaffected, which indicates that the biochemical differences essentially depended on enzyme/substrate affinities. On the whole, vaccenic and elaidic acid were shown to incorporate cell phospholipids unequally, at least in vitro, which suggests they can differently affect lipid metabolic pathways in normal cells.     

Regiospecific analysis of triacylglycerols

Isobaric triacylglycerols which differ in the sn‐1(3) and sn‐2 positions of acyl chains are regioisomers. The objective of this review is to describe the resolution of regioisomers by silver‐ion chromatography, RP–HPLC, and by GLC. Furthermore, the aim of this review is to discuss the determination of regioisomers solely by mass spectrometry and by chromatography‐mass spectrometry (GLC and electron ionization MS, silver‐ion chromatography and APCI–MS or ESI–MS, RP–HPLC and APCI–MS, and NP–HPLC and ESI–MS).     

鲨鱼皮明胶

鲨鱼皮极易被酸或碱膨胀,所得产品明胶的冻力平均大于200Bloom-g,平均黏度高达9mPa.s(相当30°E)。     

Simple,Rapid Lipidomic Analysis of Triacylglycerols in Bovine Milk by Infusion-Electrospray Mass Spectrometry

Bovine milk is a complex mixture of lipids, proteins, carbohydrates, and other factors of which lipids comprise 3–5% of the total mass. Rapid analysis and characterization of the triacylglycerols (TAG) that comprise about 95% of the total lipid is daunting given the numerous TAG species. In the attached methods paper, we demonstrate an improved method for identifying and quantifying TAG species by infusion-based “shotgun” lipidomics. Because of the broad range of TAG species in milk, a single internal standard was insufficient for the analysis and required sectioning the spectrum into three portions based upon mass range to provide accurate quantitation of TAG species. Isobaric phospholipid interferences were removed using a simple dispersive solid-phase extraction step. Using this method, > 100 TAG species were quantitated by acyl carbon number and desaturation level in a sample of commercially purchased bovine milk.     

Triacylglycerols of evening primrose oenothera biennis seed oil

The triacylglycerol (TG) composition of evening primrose(Oenothera biennis) seed oil (EPO) was studied using a combination of silver nitrate thin layer chromatography (AgNO```3``-TLC), reverse phase high performance liquid chromatography (HPLC) and capillary gas liquid chromatography (GLC). The important TGs in EPO are LLL (24.4%), LLO (23.9%), LLP (11.5%), LOO (7.2%), LOP (6.8%), LLS (4.8%), γLnLp (3.7%), LOS (3.3%), γLnLS (2.0%), γLnLL (2.0%), LPP (1.9%), OOO (1.7%), LSP (1.3%) and γLnLO (1.0%).     

Analysis of Triacylglycerols and Free Fatty Acids in Algae Using Ultra-Performance Liquid Chromatography Mass Spectrometry

To assess the suitability of microalgal strains for biodiesel production the lipid content and composition, especially individual triacylglycerols (TAG) and free fatty acids (FFA) must be determined. In this study, the compositions and concentrations of TAG and FFA were analysed in four halophytic algal species, Dunaliella salina, D. tertiolecta, D. bardawil, and D. granulata. These species were selected as part of a larger screen to identify species suitable for biofuel feedstocks. An accelerated solvent extraction instrument was used for lipids and fatty acid extraction using a dichloromethane–hexane solvent system. Ultra-performance liquid chromatography coupled with mass spectrometry (MS) detection was optimized and applied to the quantitative analysis of TAG and FFA in the different algal extracts. Individual TAG were characterized structurally using direct electrospray ionization (ESI) MS and MS/MS techniques. Cationic adducts (NH₄ ⁺) of TAG were detected and quantified in the positive ESI MS and MS/MS modes, while the negative ESI mode was used for FFA analysis. Over 20 TAG were identified and quantified in the four Dunaliella strains. Analysis of FFA compositions demonstrated that the most abundant FFA in these four algal species were palmitic, linolenic, linoleic, and oleic acids.     

Rapid Preparation of Fluorescent 9-Anthrylmethyl Esters for Fatty Acid Analysis of Small Amount of Triacylglycerols

This paper proposes a one-step method for preparation of fluorescent 9-anthrylmethyl esters from triacylglycerols (TAG) ranging in amount from 0.1 to 5 μg. It involves base-catalyzed transesterification using potassium 9-anthracenemethoxide, prepared by proton exchange between 9-anthracenemethanol and potassium tert-butoxide. Transesterification for 10 min at room temperature gave the fatty acid 9-anthrylmethyl esters in nearly maximal yields (82–85%). The products could be analyzed by reversed-phase HPLC without purification. Excellent linear relationships were observed for standard curves of 10–250 pmol of TAG standards (16:0, 19:0, 18:2 and 22:6), and differences in the slopes were less than 5% among the standards. Almost consistent compositions of the esters were observed for the products formed from 0.5 to 5 μg or less of fish oils TAG, and they were similar to those obtained by HPLC of ordinary multi-step synthesis products and by GLC of methyl esters. The present method is a great improvement of derivatization time, and is powerful for fatty acid analysis of small amounts of natural TAG.     

Medium- and Long-Chain Triacylglycerols Reduce Body Fat and Blood Triacylglycerols in Hypertriacylglycerolemic, Overweight but not Obese, Chinese Individuals

In contrast to the consumption of long-chain triacylglycerols (LCT), consumption of medium- and long-chain triacylglycerols (MLCT) reduces the body fat and blood triacylglycerols (TAG) level in hypertriacylglycerolemic Chinese individuals. These responses may be affected by BMI because of obesity-induced insulin resistance. We aimed to compare the effects of consuming MLCT or LCT on reducing body fat and blood TAG level in hypertriacylglycerolemic Chinese subjects with different ranges of BMI. Employing a double-blind, randomized and controlled protocol, 101 hypertriacylglycerolemic subjects (including 67 men and 34 women) were randomly allocated to ingest 25–30 g/day MLCT or LCT oil as the only cooking oil for 8 consecutive weeks. Anthropometric measurements of body weight, BMI, body fat, WC, HC, blood biochemical variables, and subcutaneous fat area and visceral fat area in the abdomen were measured at week 0 and 8. As compared to subjects with BMI 24–28 kg/m² in the LCT group, corresponding subjects in the MLCT group showed significantly greater decrease in body weight, BMI, body fat, WC, ratio of WC to HC, total fat area and subcutaneous fat area in the abdomen, as well as blood TAG and LDL-C levels at week 8. Based upon our results, consumption of MLCT oil may reduce body weight, body fat, and blood TAG and LDL-C levels in overweight hypertriacylglycerolemic Chinese subjects but may not induce these changes in normal or obese hypertriacylglycerolemic subjects.     

Oxidative Stability of Polyunsaturated Triacylglycerols Encapsulated in Oleaginous Yeast

Oleaginous yeast cells have the ability to synthesize oil from carbon sources or to adsorb fatty acids from their growth medium. Fish oil or conjugated linoleic acid (CLA)-rich oils encapsulated in Cryptococcus curvatus were protected from oxidation for more than 7 weeks. Oil-containing dead and viable yeast as well as oils extracted from dead or viable yeast were incubated at 52 °C in the dark. Oils extracted from yeast at the beginning of the experiment began oxidizing almost immediately and exceeded peroxide values (PV) of 20 mequiv/kg within a few days and eventually reaching PV > 100 mequiv/kg. After 56 days of incubation the PV value of oil from viable cells grown on fish oil was 3.8 ± 0.1 and 5.5 ± 0.8 mequiv/kg from dead cells. After 42 days of incubation the PV of oil from viable CLA containing yeast was 1.1 ± 0.2 mequiv/kg and 1.7 ± 0.5 from dead CLA containing yeast. C. curvatus encapsulation significantly improved oxidative stability of long-chain polyunsaturated fatty acids (LCPUFA) and CLA. Yeast cell viability was not critical for oxidative stability of the encapsulated oil.     

Study of Some Experimental Parameters in the Synthesis of Triacylglycerols with CLA Isomers and Structural Analysis

The present research deals with the synthesis of structured triacylglycerols (TAG) by enzymatic treatment of sn-1,3-diacylglycerol (sn-1,3-DAG) with conjugated linoleic acid (CLA) isomers using the immobilized lipase from Rhizomucor miehei (Lipozyme^® IM) under different experimental conditions. In particular, the influence of reaction parameters, such as temperature, enzymatic load, reaction time and DAG/CLA ratio has been evaluated using an experimental design software with a screening objective. Two responses have been selected, they are the percentage of CLA isomers in total TAG and in the sn-2- position and a three-level-4-factor fractional factorial experimental design was used to screen the variables. The results showed that the selected experimental variables have an influence on the enzymatic reaction, in particular, the DAG/CLA substrate ratio and the temperature, both of which inversely correlated with CLA incorporation, but also the enzymatic load and the reaction time, both directly correlated with CLA incorporation. The best results for CLA isomer % content both in total TAG (46.3%) and in the sn-2- position (52.2%) were obtained at 40 °C for 96 h, with 20% enzymatic load and a 0.5 reactive ratio.     

Synthesis of Symmetrical Diacid Triacylglycerols from Glycerol-1,3-Ditosylate

The tosylate route has been explored for the preparation of symmetrical diacid triacylglycerols. In this route best conditions have been found for the preparation of glycerol-1,3-ditosylate and its subsequent conversion into 1,3-diacylglycerols by nucleophilic substitution. From the 1,3-diacylglycerols, five symmetrical diacid triacylglycerols have been prepared reaction with an appropriate acid chloride.     

Regiospecific analysis of triacylglycerols using allyl magnesium bromide

A method for the regiospecific analysis of triacylglycerols (TAG), using the Grignard reagent allyl magnesium bromide (AMB) to partially deacylate TAG, is described. 1,3-Distearoyl-2-oleoyl-glycerol (SOS) and 1,3-didecanoyl-2-palmitoyl-glycerol (CPC) were reacted with AMB. From the resulting mixture, the four different classes of partial acylglycerols and TAG were isolated, and the mole ratios between stearic acid and oleic acid, or decanoic acid and palmitic acid, respectively, were determined in each fraction. Different approaches of calculating the composition of the fatty acids in positionssn-1(3) andsn-2 of the original TAG were compared. For thesn-2 position, the best estimate was the direct determination of the fatty acid composition of 2-monoacylglycerol (MAG). Mole percentages of stearic acid and decanoic acid in thesn-1(sn-3) positions of SOS and CPC, respectively, were most accurately estimated from the fatty acid compositions of TAG and 2-MAG according to the formula: 1.5×TAG−0.5×2-MAG. Using AMB and the present method of calculation, the results obtained were more accurate and showed smaller standard derivations than those obtained using other common deacylating agents, such as ethyl magnesium bromide or pancreatic lipase.     

Structure and Biological Significance of Triacylglycerols Containing Cyclopropene Acyl Moieties

Cyclopropene fatty acids are constituents of seed lipids of various plants, including sources of edible fats, such as cottonseed and kapok. Physiological and toxicological properties of lipids containing cyclopropene acyl moieties are reviewed and the difficulties encountered in the elucidation of the structures of triacylglycerols containing cyclopropene acyl moieties are discussed. A novel method for the analysis of such triacylglycerols is presented.     

Separation of Enantiomeric Triacylglycerols by Chiral‐Phase HPLC

Liquid chromatography–atmospheric pressure chemical ionization mass spectrometry (LC–MS/APCI) with a chiral phase was used for separation of triacylglycerols (TAG) obtained either by organic synthesis or isolated from different algal species. We present chromatographic characteristics and tandem mass spectra of enantiomers and positional isomers (regioisomers) of C16, C18 and C20 polyunsaturated fatty acids. The retention time was found to depend on the structure of the given TAG, increasing with increasing number of double bonds and decreasing with increasing number of the carbons in TAG, with the exception of dieicosapentaenoyl‐palmitoyl‐glycerols.     

Triacylglycerols and regiospecific fatty acid analyses of philippine seed oils

Three Philippine seed oils, namely coconut (Cocos nucifera Linn.), pilinut (Canarium ovatum Engl.), and cashew (Anacardium occidentale Linn.), which were selected for their local abundance and availability, were examined for their triacylglycerol profiles and fatty acid compositions. Triacylglycerol molecular species in terms of carbon number and partition number were determined by gas chromatography and liquid chromatography, respectively. The distribution of fatty acids in the primary and secondary positions of the glycerol backbones for the three oils were examined by regiospecific analysis by using pancreatic lipase. Coconut oil had high concentrations of lauric and myristic acids, while the other two oils did not have such fatty acids. Lauric acid in coconut oil and linoleic acid in pilinut oil were distributed mainly in the primary positions (sn-1,3) of the glycerol backbone. Trilaurin and dioleylpalmitoylglycerol were the major triglycerides in coconut and pilinut oils, respectively.     

Glass Transition and Water Activity of Freeze-Dried Shark

Glass transition temperatures and water activity as a function of moisture content were determined for freeze-dried shark to further compare the two distinct criteria of food stability. The adsorption experiments were carried out at controlled water activity, at 21, 40, and 50°C using an isopiestic method and they were modeled using BET and GAB equations. Glass transition temperatures were measured by dynamic oscillation on shear. Results indicate that there is a difference in the temperature-related stability criteria predicted by the concepts of water activity (a _w ) and the glass phenomenon (T _g ). The glass transition concept tends to accommodate higher levels of moisture within the framework of a safe storage of a dried product.     

Lipidomic Profiling of Chylomicron Triacylglycerols in Response to High Fat Meals

Using lipidomic methodologies the impact that meal lipid composition and metabolic syndrome (MetS) exerts on the postprandial chylomicron triacylglycerol (TAG) response was examined. Males (9 control; 11 MetS) participated in a randomised crossover trial ingesting two high fat breakfast meals composed of either dairy-based foods or vegetable oil-based foods. The postprandial lipidomic molecular composition of the TAG in the chylomicron-rich (CM) fraction was analysed with tandem mass spectrometry coupled with liquid chromatography to profile CM TAG species and targeted TAG regioisomers. Postprandial CM TAG concentrations were significantly lower after the dairy-based foods compared with the vegetable oil-based foods for both control and MetS subjects. The CM TAG response to the ingested meals involved both significant and differential depletion of TAG species containing shorter- and medium-chain fatty acids (FA) and enrichment of TAG molecular species containing C16 and C18 saturated, monounsaturated and diunsaturated FA. Furthermore, there were significant changes in the TAG species between the food TAG and CM TAG and between the 3- and 5-h postprandial samples for the CM TAG regioisomers. Unexpectedly, the postprandial CM TAG concentration and CM TAG lipidomic responses did not differ between the control and MetS subjects. Lipidomic analysing of CM TAG molecular species revealed dynamic changes in the molecular species of CM TAG during the postprandial phase suggesting either preferential CM TAG species formation and/or clearance.