Bovine Milk Triacylglycerol Regioisomer Ratio Shows Remarkable Inter-Breed and Inter-Cow Variation.

Regioisomers (or positional isomers) of triacylglycerols (TAGs) of milk are known to show differential outcome in relation to human absorption. Quantitation of TAG regioisomers remains a big challenge due to the lack of facile chromatographic separation technique. The feasibility of using fragment ion intensity ratio to determine the ratio of co-eluting AAB/ABA-type regioisomer pairs was confirmed in this study. The ability of C30 stationary phase in resolving interfering TAG isomers was demonstrated for the first time. This allowed us to reveal the complexity of using fragment ion intensity to quantify 1,2-olein-3-palmitin (OOP), 1,3-olein-2-palmitin (OPO), 1,2-olein-3-stearin (OOS), and 1,3-olein-2-stearin (OSO) regioisomers in milk samples. A novel algorithm was proposed to consider the contribution of OPO/OOP and OSO/OOS double bond (DB)-isomers and to eliminate the interference of isobaric ions from other isomers, an aspect overlooked in previous studies. This liquid chromatography-mass spectrometry method that requires no pre-fractioning and a moderate chromatographic separation time of 36 min is simple and, thus, suitable for screening a large number of samples for genetic analysis of this trait. Preliminary results using a small cohort of animals showed that OPO/OOP ratio differs significantly between Jersey and Holstein cows, and a large variation was also observed across individual Holstein cows.

Lipid profile of bovine grade-1 blastocysts produced either in vivo or in vitro before and after slow freezing process.

Currently, in vitro embryo production (IVP) is successfully commercially applied in cattle. However, the high sensitivity of embryos to cryopreservation in comparison to in vivo (IVD) embryos slows the dissemination of this biotechnology. Reduced cryotolerance is frequently associated with lipid accumulation in the cytoplasm mainly due to in vitro culture conditions. The objective of this study was to evaluate the lipid composition of biopsied and sexed embryos, produced either in vivo or in vitro from the same Holstein heifers before and after a slow freezing protocol. Lipid extracts were analysed by liquid chromatography-high resolution mass spectrometry, which enabled the detection of 496 features. Our results highlighted a lipid enrichment of IVP embryos in triglycerides and oxidised glycerophospholipids and a reduced abundance in glycerophospholipids. The slow freezing process affected the lipid profiles of IVP and IVD embryos similarly. Lysophosphatidylcholine content was reduced when embryos were frozen/thawed. In conclusion, the embryonic lipid profile is impacted by IVP and slow freezing protocols but not by sex. Lysophosphatidylcholine seemed highly sensitive to cryopreservation and might contribute to explain the lower quality of frozen embryos. Further studies are required to improve embryo freezability by modulating the lipidome.

Obese Mice with Dyslipidemia Exhibit Meibomian Gland Hypertrophy and Alterations in Meibum Composition and Aqueous Tear Production.

BACKGROUND: Dyslipidemia may be linked to meibomian gland dysfunction (MGD) and altered meibum lipid composition. The purpose was to determine if plasma and meibum cholesteryl esters (CE), triglycerides (TG), ceramides (Cer) and sphingomyelins (SM) change in a mouse model of diet-induced obesity where mice develop dyslipidemia. METHODS: Male C57/BL6 mice (8/group, age = 6 wks) were fed a normal (ND; 15% kcal fat) or an obesogenic high-fat diet (HFD; 42% kcal fat) for 10 wks. Tear production was measured and meibography was performed. Body and epididymal adipose tissue (eAT) weights were determined. Nano-ESI-MS/MS and LC-ESI-MS/MS were used to detect CE, TG, Cer and SM species. Data were analyzed by principal component analysis, Pearson's correlation and unpaired t-tests adjusted for multiple comparisons; significance set at p ≤ 0.05. RESULTS: Compared to ND mice, HFD mice gained more weight and showed heavier eAT and dyslipidemia with higher levels of plasma CE, TG, Cer and SM. HFD mice had hypertrophic meibomian glands, increased levels of lipid species acylated by saturated fatty acids in plasma and meibum and excessive tear production. CONCLUSIONS: The majority of meibum lipid species with saturated fatty acids increased with HFD feeding with evidence of meibomian gland hypertrophy and excessive tearing. The dyslipidemia is associated with altered meibum composition, a key feature of MGD.

Determination of biodiesel and used cooking oil in automotive diesel/green diesel fuels through high-performance liquid chromatography.

This study reports a simple and convenient analytical method for the simultaneous determination of biodiesel and vegetable oils or used cooking oils in petrodiesel and green diesel (hydrotreated vegetable oils or paraffinic diesel). The approach is based on normal-phase high-performance liquid chromatography with refractive index detection. It employed silica stationary phase, n-hexane mobile phase with isopropanol modifier to achieve optimum separation between hydrocarbons (petrodiesel or green diesel), fatty acid methyl esters (biodiesel) and triglycerides (vegetable oils and used cooking oil). In addition to determining vegetable oils or used cooking oils as adulterants in diesel, this method is also proposed as a better alternative to the standard method ASTM D7371, which is currently recommended for determining fatty acid methyl esters in petrodiesel. The method development involved screening of various stationary and mobile phases, with and without modifiers, to achieve acceptable chromatographic resolutions between analytes. Under the optimized method conditions, silica column, and n-hexane containing 0.6% isopropanol as the mobile phase provided the best results. The real-world scenario was simulated for the method validation carried out by fortifying Jatropha seed oil, soybean oil, and used cooking oil in the biodiesel blended petrodiesel and green diesel. Measurement of all analytes was accompanied by high precision, low limit of detection/quantification and linear response range of 0.05 to 50% for biodiesel, and 0.05 to 30% for vegetable oils. The proposed method is simple, fast (runtime 7 min), and does not require sample pre-treatment and backflushing.

Multidimensional gas chromatographic‒Mass spectrometric method for separation and identification of triacylglycerols in olive oil.

A 'heart-cut' multidimensional gas chromatography‒mass spectrometry (H/C MDGC‒MS) method for separation and identification of triacylglycerols (TAGs) in extra virgin olive oil was developed. A GC configuration, comprising a non-polar first dimension (1D) column (15 m length) and a mid-polarity second dimension (2D) column (9 m length), was employed. Standard TAGs were used to test and demonstrate the H/C MDGC method, for identification of TAG components and to validate the method. Various chromatographic conditions such as column flow and temperature program were evaluated. The 1D separation resulted in overlap of some standard TAG peaks. These overlapped 1D regions of the standard TAGs were H/C to 2D for further separation and resulted in clearly distinguished individual TAG component peaks. The 1D separation of olive oil TAGs displayed three major peaks and four minor peaks. The application of the H/C MDGC method to olive oil TAGs resulted in the separation of each sampled 1D region into two or more TAG peaks. TAG components in olive oil resolved on the 2D column were identified based on characteristic mass fragment ions such as [M-RCO2]+, [RCO+128]+, [RCO+74]+ and RCO+ and comparison of their mass spectra with that of the standard TAGs. Sixteen olive oil TAGs were identified by MS after 2D separation. The repeatability of the H/C method was evaluated in terms of retention time shift and area response in the 2D and found to be <0.02% and <8% RSD respectively.

Thladiantha Seed Oils - New Source of Conjugated Fatty Acids: Characterization of Triacylglycerols and Fatty Acids.

In this study, seed oils of Thladiantha nudiflora and Thladiantha dubia were found to contain 55.5 and 44.4% mole of conjugated octadecatrienoic fatty acids, respectively. The presence of moieties of conjugated fatty acids was confirmed by a series from physical methods: UV, IR, 1H and 13C NMR. The triacylglycerols (TAGs) isolated of the seed oils were studied by RP-HPLC with diode array and mass spectrometric detections. It was shown that all 15 TAGs of Thladiantha dubia contain moieties of conjugated fatty acids - punicic, (9Z,11E,13Z)-octadeca-9,11,13-trienoic acid (35.6% mole) and 8.9% mole α-eleostearic, (9Z,11E,13E)-octadeca-9,11,13-trienoic acid. Meanwhile, 24 TAGs of Thladiantha nudiflora seed oil contain both acids in approximately equal proportions (27.4:28.2 % mole). The enrichment for polyunsaturated fatty acids of the hydrolysis product of the seed oils due to urea inclusion complex formation was discussed.

Enzymatic Preparation and Oxidative Stability of Human Milk Fat Substitute Containing Polyunsaturated Fatty Acid Located at sn-2 Position.

The development of human milk fat substitutes (HMFSs), rich in palmitic acid (16:0) at the sn-2 position of triacylglycerol (TAG) and rich in unsaturated fatty acids (FAs) (oleic acid, 18:1 and linoleic acid, 18:2) at the sn-1(3) positions, has gained popularity. In this study, HMFSs containing polyunsaturated fatty acids (PUFAs) predominantly at the sn-2 position were prepared, and their oxidation stabilities were compared. First, a non-PUFA-containing HMFS (NP-HMFS) was produced by enzymatic reactions using Novozyme® 435 and Lipozyme® RM-IM as the enzymes and lard as the raw material. Second, HMFSs, containing 10 % PUFA at the sn-2 or sn-1(3) position, were individually prepared by enzymatic reactions using lard and fish oil as raw materials. Here, sn-2-PUFA-monoacylglycerol (MAG) was extracted from the reaction solution using a mixture of hexane and ethanol/water (70:30, v/v) to produce high-purity sn-2-PUFA-MAG with 78.1 % yield. For the PUFA-containing HMFS substrates, comparable oxidation stability was confirmed by an auto-oxidation test. Finally, HMFSs containing 10 % or 2 % sn-1,3-18:1-sn-2-PUFA-TAG species were prepared by enzymatic reactions and subsequent physical blending. The oxidative stability of sn-1,3-18:1-sn-2-PUFA-HMFS was two-fold higher than that of 1/2/3-PUFA-HMFS in which each PUFA was located without stereospecific limitations in TAG. The removal of PUFA-TAG molecular species with higher concentrations of unsaturated units had a significant effect. In addition, the oxidation stability increased with the addition of tocopherol as an antioxidant. Thus, the combined use of two strategies, that is, the removal of PUFA-TAG molecular species with high concentrations of unsaturated units and the addition of antioxidants, would provide a PUFA-containing HMFS substrate with high oxidative stability.

miRNA/Target Gene Profile of Endothelial Cells Treated with Human Triglyceride-Rich Lipoproteins Obtained after a High-Fat Meal with Extra-Virgin Olive Oil or Sunflower Oil.

SCOPE: The effects of triglyceride-rich lipoproteins (TRLs) on the miRNA expression of endothelial cells, which are very involved in atherosclerosis, according to the type of diet are not known. METHODS AND RESULTS: The differences between the effects of TRLs isolated from blood of subjects after a high-fat meal with extra-virgin olive oil (EVOO) and sunflower oil (SO) on the microRNA-Seq profile related to atherosclerosis in human umbilical vein endothelial cells are analyzed. 28 upregulated microRNAs with EVOO-derived TRLs, which can regulate 22 genes related to atherosclerosis, are found. 21 upregulated microRNAs with SO-derived TRLs, which can regulate 20 genes related to atherosclerosis, are found. These microRNAs are mainly involved in angiogenesis, with a predominance of an anti-angiogenic effect with EVOO-derived TRLs. Other microRNAs upregulated with SO-derived TRLs are involved in cardiovascular diseases. Pathways for the target genes obtained from the upregulated microRNA with EVOO-derived TRLs are involved in lipid metabolism and inflammatory and defense response, while those with SO-derived TRLs are involved in lipid metabolic process. CONCLUSION: EVOO-derived TRLs seem to produce a more atheroprotective profile than SO-derived TRLs. This study provides alternative mechanisms on the protective role of EVOO against the atherogenic process through microRNA regulation in endothelial cells.

Triacylglycerols Profiles Established by UHPLC-ESI-MS in Developing Sweet, Semi-sweet and Bitter Seeds from Tunisian Oilseeds Apricot (Prunus armeniaca L.).

The aim of the present research is to investigate the effect of three harvest date on the composition of apricot seed. Indeed, triacylglycerols (TAGs) content and composition were studied in developing Tunisian apricot varieties bitter (Bargoug), semi-sweet (Oud Rhayem) and sweet (Chechi Bazza) cultivars at intervals of early (14 DAP), mid phase (28 DAP) and full phase (55 DAP) of oil accumulation by UHPLC-ESI-MS method. Eleven molecular species of triacylglycerols were detected and identified as LLL, LLO, LLP, LOO, LLS/LOP, LPP, OOO, LOS, OOP, POP and OOS. At 14 DAP, LLO was the major TAGs molecular species with 35.4-52.6% (maximum reached in semi-sweet apricot). Others major TAGs were founded at lower content as LOO (17.5-40.3%) and OOO (5.7-12.7%). However, among maturity, three distinct profiles of TAGs molecular species were observed: bitter apricot was significantly richer in OOO molecular species than cultivars ones. However, semi-sweet and sweet cultivars were richer in LLO and LOO molecular species at different time-dates. These latter may provide a schedule for harvesting Tunisian apricot seeds with high quality of oil content.

Separation of triacylglycerols containing allenic and acetylenic fatty acids by enantiomeric liquid chromatography-mass spectrometry.

Triacylglycerols (TAGs) containing less common fatty acids (FAs) were isolated from the seeds of three plants (Santalum album, Crepis foetida, and Leucas aspera). These FAs had allenic (laballenic acid, Lb) and acetylenic (crepenynic, C; ximenynic acids, Xi) bonds. TAGs were analyzed on reversed-phase and chiral columns. High-resolution tandem mass spectrometry identified TAGs by positive electrospray ionization (ESI+). Twenty-two molecular species of TAGs isolated from the seed oil of Santalum album were separated by RP-HPLC and chiral HPLC methods and identified by positive electrospray ionization tandem MS detection (ESI+-MS). Two major enantiomers, i.e., sn-OOLb and sn-LLLb (O represents oleic acid; and L represents linoleic acid), were synthesized from the appropriate phosphatidylcholines. This allowed the identification of enantiomers after separation by chiral chromatography by tandem mass spectrometry. Similarly, TAGs from the seeds of Crepis foetida, and Leucas aspera were analyzed by reversed-phase chromatography and identified by mass spectrometry. Four enantiomers (sn-OOC, sn-LLC, sn-OOXi, and sn-LLXi) were synthesized. A total of six and three enantiomers of TAGs containing crepenynic and ximenynic acids, respectively, were identified by chiral column analysis. The retention times of TAGs containing allenic and acetylenic bonds were always greater on the reversed-phase column than TAGs with the same number of carbon atoms and the same unsaturation (e.g., LLL versus LLLb). From the chiral column, the regioisomers and enantiomers were eluted in the order of symmetric-asymmetric-asymmetric (i.e., sn-OCO, sn-COO, and sn-OOC). Through tandem mass spectrometry, we were able to identify and distinguish regioisomer [DAG]+-type ions, i.e., [MNH4NH3RCOOH]+, that can be considered diagnostic. Unfortunately, enantiomers and TAGs with the same numbers of carbon atoms and the same unsaturation levels have identical mass spectra, such as LLL and LLLb.

Lipidomic characterization of exosomes isolated from human plasma using various mass spectrometry techniques.

Ultrahigh-performance supercritical fluid chromatography - mass spectrometry (UHPSFC/MS), ultrahigh-performance liquid chromatography - mass spectrometry (UHPLC/MS), and matrix-assisted laser desorption/ionization (MALDI) - MS techniques were used for the lipidomic characterization of exosomes isolated from human plasma. The high-throughput methods UHPSFC/MS and UHPLC/MS using a silica-based column containing sub-2 µm particles enabled the lipid class separation and the quantitation based on exogenous class internal standards in <7 minute run time. MALDI provided the complementary information on anionic lipid classes, such as sulfatides. The nontargeted analysis of 12 healthy volunteers was performed, and absolute molar concentration of 244 lipids in exosomes and 191 lipids in plasma belonging to 10 lipid classes were quantified. The statistical evaluation of data included principal component analysis, orthogonal partial least square discriminant analysis, S-plots, p-values, T-values, fold changes, false discovery rate, box plots, and correlation plots, which resulted in the information on lipid changes in exosomes in comparison to plasma. The major changes were detected in the composition of triacylglycerols, diacylglycerols, phosphatidylcholines, and lysophosphatidylcholines, whereby sphingomyelins, phosphatidylinositols, and sulfatides showed rather similar profiles in both biological matrices.

Separation of postprandial lipoproteins: improved purification of chylomicrons using an ApoB100 immunoaffinity method.

Elevated levels of triglyceride-rich lipoproteins (TRLs), both fasting and postprandial, are associated with increased risk for atherosclerosis. However, guidelines for treatment are defined solely by fasting lipid levels, even though postprandial lipids may be more informative. In the postprandial state, circulating lipids consist of dietary fat transported from the intestine in chylomicrons (CMs; containing ApoB48) and fat transported from the liver in VLDL (containing ApoB100). Research into the roles of endogenous versus dietary fat has been hindered because of the difficulty in separating these particles by ultracentrifugation. CM fractions have considerable contamination from VLDL (purity, 10%). To separate CMs from VLDL, we produced polyclonal antibodies against ApoB100 and generated immunoaffinity columns. TRLs isolated by ultracentrifugation of plasma were applied to these columns, and highly purified CMs were collected (purity, 90-94%). Overall eight healthy unmedicated adult volunteers (BMI, 27.2 ± 1.4 kg/m2; fasting triacylglycerol, 102.6 ± 19.5 mg/dl) participated in a feeding study, which contained an oral stable-isotope tracer (1-13C acetate). We then used this technique on plasma samples freshly collected during an 8 h human feeding study from a subset of four subjects. We analyzed fractionated lipoproteins by Western blot, isolated and derivatized triacylglycerols, and calculated fractional de novo lipogenesis. The results demonstrated effective separation of postprandial lipoproteins and substantially improved purity compared with ultracentrifugation protocols, using the immunoaffinity method. This method can be used to better delineate the role of dietary sugar and fat on postprandial lipids in cardiovascular risk and explore the potential role of CM remnants in atherosclerosis.

Antidiabetic components from the hexane extract of red kidney beans (Phaseolus vulgaris L.): isolation and structure determination.

Red kidney beans (Phaseolus vulgaris L.) contain bioactive compounds that are known to exhibit antidiabetic effects via inhibition of α-glucosidase. However, information on the nonpolar components that exhibit antidiabetic activity is limited. Here, we report the isolation and structure determination of components with α-glucosidase inhibitory activity, which were obtained from the hexane extract of red kidney beans. Triacylglycerols (TAGs) were identified as the major components exhibiting inhibitory activity against α-glucosidase. The chemical structure of TAGs was determined by a combination of GC-MS and UPLC-MS/MS. The primary TAGs identified were LnLnLn (trilinolenin) and LnLLn (1,3-dilinolenoyl-2-linoleoyl glycerol). The major fatty acids present in these TAGs were α-linolenic acid (ω-3) and linoleic acid (ω-6). These TAGs were also found to inhibit the α-glucosidase activity in a similar fashion as acarbose. These results suggest that TAGs have potency as antidiabetics and support the potential suitability of red kidney beans for diabetes treatment.

Relationship between gut microbiota and circulating metabolites in population-based cohorts.

Gut microbiota has been implicated in major diseases affecting the human population and has also been linked to triglycerides and high-density lipoprotein levels in the circulation. Recent development in metabolomics allows classifying the lipoprotein particles into more details. Here, we examine the impact of gut microbiota on circulating metabolites measured by Nuclear Magnetic Resonance technology in 2309 individuals from the Rotterdam Study and the LifeLines-DEEP cohort. We assess the relationship between gut microbiota and metabolites by linear regression analysis while adjusting for age, sex, body-mass index, technical covariates, medication use, and multiple testing. We report an association of 32 microbial families and genera with very-low-density and high-density subfractions, serum lipid measures, glycolysis-related metabolites, ketone bodies, amino acids, and acute-phase reaction markers. These observations provide insights into the role of microbiota in host metabolism and support the potential of gut microbiota as a target for therapeutic and preventive interventions.

Effect of Fractionation and Chemical Characteristics on the Crystallization Behavior of Milk Fat.

The experiments reported in this study provided a more comprehensive insight into the effect of chemical composition on the crystallization behavior of milk fat (MF). MF was fractionated between 20 and 40 °C into nine fractions with different melting points and was first subjected to the heating step (L20, L30, L40, and S40) followed by the cooling phase (SS40, SL40, SS30, SL30, and LL40). Furthermore, the species of fatty acids (FAs) and triglycerides (TAGs) of the MF fractions were identified. The thermodynamics, crystallization behavior, and polymorphs were determined using differential scanning calorimetry, pulsed nuclear magnetic resonance, and X-ray diffraction, respectively. The results indicated that L40 yielded the highest percentage (∼35% of the total MF) of all the fractions. Enthalpies of the melting and crystallization processes of solid fat content in this study were related to the different FA and TAG compositions of MF and its fractions. High melting fractions (HMFs) were enriched with long-chain saturated fatty acids and tri-saturated (S3) TAGs, and low melting fractions (LMFs) were enriched with short-chain unsaturated FAs and tri-unsaturated (U3) TAGs. Moreover, the various nucleation mechanisms of MF fractions were identified according to the Avrami equation. The polymorphic transformation from a ß' form of double chain length structures to a ß form of triple chain length occurred in the native MF and HMFs, whereas the LMFs displayed almost no crystals. PRACTICAL APPLICATION: This study represented the first time that nine fractions were obtained using MF fractionation via a heating step, followed by a cooling phase. Furthermore, the chemical composition of MF fractions was investigated. The results obtained from this study might be of specific value in understanding the functional properties of fat-based dairy food in both storage conditions and real-time applications.

Effect of Heating on Compositional Characteristics and Oxidative Stability of Crude and Refined Rice Bran Oil.

The compositional characteristics and oxidative stability of rice bran oil were determined by observing the formation of oxidative products and alteration in chemical composition of oils during microwave or oven heating. The values of oxidative indicators such as free acidity, peroxide, p-anisidine, total oxidation, thiobarbituric acid and color values, increased faster in refined oils compared to crude ones during heating. In gas chromatography analysis, the percentages of total saturated, monounsaturated and polyunsaturated fatty acids in the studied oils such as lab extracted crude rice bran oil, lab extracted and refined rice bran oil, crude rice bran oil from commercial mill and refined rice bran oil from commercial mill were: 23.07 to 23.56, 41.15 to 42.38 and 34.38 to 35.88, respectively. The heating caused the reduction of polyunsaturated fatty acids content with increasing saturated fatty acids content, and these changes were greater in refined rice bran oil indicating extensive lipid oxidation occurred in refined oil. The change in triacylglycerol species content as determined by High-performance liquid chromatography, was lower in crude oil; the higher stability of these species in crude oil could have contribution to reduce oxidation. During thermal treatment, the generation of hydroperoxides, their degradation and formation of secondary oxidative products evaluated by Fourier-transform infrared spectroscopy, were lower in crude oils. However, the rate of formation of oxidative products in lab prepared samples was lower compared to that in the samples collected from commercial mill. Under extreme thermal condition, the order of oxidative stability: lab extracted crude rice bran oil > crude rice bran oil from commercial mill>lab extracted and refined rice bran oil > refined rice bran oil from commercial mill. The present results will be useful to oil seed processing mills in refining of rice bran oil for economic feasibility and better marketability.

Simultaneous Separation of Triacylglycerol Enantiomers and Positional Isomers by Chiral High Performance Liquid Chromatography Coupled with Mass Spectrometry.

The rapid and simultaneous separation of triacylglycerol (TAG) enantiomers and positional isomers was achieved using chiral high performance liquid chromatography (HPLC). TAGs composed of two fatty acids, which were both saturated (P: palmitic acid or S: stearic acid) and unsaturated (O: oleic acid or L: linoleic acid; e.g., sn-PPO/sn-OPP/sn-POP: 1,2-dipalmitoyl-3-oleoyl-sn-glycerol/1-oleoyl-2,3-dipalmitoyl-sn-glycerol/1,3-dilpalmitoyl-2-oleoylglycerol), were resolved into three peaks using CHIRALPAK IF-3 without recycling on the HPLC system. For example, the mixture of sn-PPO/sn-OPP/sn-POP was resolved in 30 min, although it took 150 min to resolve sn-PPO/sn-OPP using CHIRALCEL OD-RH in a previous study using a recycling HPLC system. This novel chiral HPLC method was applicable for the separation of other TAG isomers, including sn-OOP/sn-POO/sn-OPO, sn-PPL/sn-LPP/sn-PLP, sn-LLP/sn-PLL/sn-LPL, sn-SSO/sn-OSS/sn-SOS, sn-OOS/sn-SOO/sn-OSO, sn-SSL/sn-LSS/sn-SLS, and sn-LLS/sn-SLL/sn-LSL. For TAGs composed of three fatty acids containing both saturated and unsaturated fatty acids, the POL isomers were not sufficiently separated but the PSO and SOL isomers were partially separated into several peaks. Their elution order could be estimated by the fragment ions generated in the ion source of the mass spectrometer. However, TAGs consisting of only saturated or unsaturated fatty acids (e.g., sn-PSP/sn-PPS/sn-SPP and sn-OLO/sn-OOL/sn-LOO) were not separated. This novel chiral HPLC method is especially applicable for the analysis of TAG composition of semi-solid fats such as palm oil.

Timed relay contact closure controlled system for parallel second dimensions in multi-dimensional liquid chromatography.

OBJECTIVE: Short-chain triacylglycerols (TAGs) in lipid extracts of biological samples are not sufficiently resolved using conventional reversed-phase separation on two C18 columns in series, or using a two-dimensional chromatographic separation with a silver ion column as the second dimension (2D). An additional dimension of separation was required. RESULTS: The hardware and software components to allow a second second-dimension (2D) separation and three total separation dimensions were developed. Two contact closure (CC) activated 4-port, 2-position valves (4P2PVs) for ultra-high performance liquid chromatography (UHPLC) were joined together and used for one of two second dimensions in comprehensive two-dimensional liquid chromatography (2D-LC) coupled to four mass spectrometers simultaneously in parallel in an LC1MS2 × (LC1MS1 + LC1MS1) = LC3MS4 configuration. A timed contact closure circuit (TCCC) controlled the two UHPLC valves, operated by repetitive CCs for the 4P2PVs. The TCCC-controlled 4P2PVs were used to direct a portion of the 1D eluent to one of the two 2D's for separation by a quaternary UHPLC system that was not allowed by the commercial 2D-LC system. The 1D separation was a non-aqueous reversed-phase HPLC instrument used for separation of TAGs; the commercial 2D-LC 2D binary UHPLC was used for silver-ion chromatography of unsaturated TAGs; and the CC-controlled second 2D was used for separation of short-chain (SC) saturated TAGs.

Microwave assisted high performance liquid chromatography for the separation of triacylglycerols in vegetable oils using an evaporative light scattering detector.

Microwave (MW) radiation was applied to perform the separation of triacylglycerols (TGs) in oil samples. The novelty of the work lies in the application of MW radiation to assist the separation of several non-polar compounds employing a totally organic mobile phase. Once the influence of the evaporative light scattering detector (ELSD) variables on the sensitivity was optimized, the TGs separation was compared conditioning the column with either a conventional HPLC or a MW oven. Contrary to previous applications in which the mobile phase contained water, the improvement in sensitivity using MW was not as significant in comparison with conventional heating but it allowed a shortening in the retention times of several TGs in about 50% respect elution at room temperature. The method was finally applied for the quantification of most common TGs in almond, tiger nut, and argan oil.

Lipid Characteristics of Camellia Seed Oil.

Camellia oleifera, C. japonica and C. sinensis are three representative crops of the genus Camellia. In this work, we systematically investigated the lipid characteristics of these seed oils collected from different regions. The results indicated significant differences in acid value (AV), peroxide value (PV), iodine value (IV), saponification value (SV) and relative density of the above-mentioned camellia seed oils (p < 0.05). The C. japonica seed oils showed the highest AV (1.7 mg/g), and the C. sinensis seed oils showed the highest PV (17.4 meq/kg). The C. japonica seed oils showed the lowest IV (79.9 g/100 g), SV (192.7 mg/g) and refractive index (1.4633) of all the oils, while the C. sinensis seed oils showed the lowest relative density (0.911 g/cm3). The major fatty acids in the camellia seed oils were palmitic acid (16:0), oleic acid (18:1) and linoleic acid (18:2); the oleic acid in C. oleifera and C. japonica seed oils accounted for more than 80% of the total fatty acids. The oleic acid levels in the C. oleifera and C. japonica oils were higher than those in the C. sinensis seed oils, while the linoleic acid levels in the former were lower than those in the latter one. Differences also exist in the triacylglycerol (TAG) composition, although the most abundant TAG molecular species in the camellia seed oils was trioleoylglycerol (OOO). Seven sterol species, squalene and α-tocopherol were detected in the camellia seed oils, however, the contents of tocopherol and unsaponifiable molecules in the C. oleifera and C. japonica seed oils were significantly lower than those in the C. sinensis seed oil. These results demonstrated that the varieties of Camellia affected the seed oil lipid characteristics.