Tentative identification and quantification of TAG core aldehydes as dinitrophenylhydrazones in autoxidized sunflowerseed oil using reversed-phase HPLC with electrospray lonization MS

The molecular species of TAG core aldehydes (aldehydes still esterified to parent molecules) were detected and quantified in dietary-quality sunflowerseed oil autoxidized for 0‐18 d at 60°C in the dark. The analyses were performed by reversed-phase HPLC with UV (358 nm) absorption or light scattering and electrospray ionization-MS (ESI/MS) detection following preparation of the dinitrophenylhydrazone derivatives. Aldehyde production, as estimated by UV and ESI/MS, increased gradually over the 18-d period following a rapid initial destruction of the core aldehydes accumulated during storage of the commercial oil at 10°C for 3 mon. The contents of hydroperoxides and hydroperoxide core aldehyde combinations were estimated to account for about 5% of total TAG, quantified as area in the chromatographic trace, after 18 d of autoxidation as estimated by an evaporative light scattering detector (ELSD). The major species of core aldehydes were tentatively identified as 9-oxononanoyl (70%)-, 12-oxo-9,10-epoxydodecenoyl (10%)-, and 13-oxo-9,11-tridecadienoyl (5%)-containing acylglycerols, plus smaller amounts of simple and mixed chain-length dialdehydes, and hydroxy and epoxy monoaldehyde-containing acylglycerols (15% of total). Quantitatively, the core aldehydes made up 2–12 g/kg of oil by UV detection and 2–9 g/kg of oil by EsI/MS detection, whereas the hydroperoxides measured in the unreduced state by HPLC with ELSD were estimated at 200 g/kg after 18 d of autoxidation. The major hydroperoxides of sunflowerseed oil were as previously identified.     

Liquid Chromatography–Light Scattering Detector–Mass Spectrometric Analysis of Digested Oxidized Rapeseed Oil

Rapeseed oil was oxidized chemically and thermally to produce two distinct oxidized oils. These oils, along with unoxidized oils, were subjected to an artificial digestion model to simulate the digestive processes in humans. Lipid digestion involves lipases that break down the intact triacylglycerol (TAG) molecules first to diacylglycerols, and eventually to sn-2-monoacylglycerols (MAG) and free fatty acids. A high performance liquid chromatography-evaporative light scattering detector-electrospray ionization-mass spectrometric (HPLC–ELSD–ESI–MS) method was developed to monitor the lipolysis and the presence of oxidized lipids. The HPLC–ELSD–ESI–MS analysis enabled the separation and detection of nearly all the lipid species present in the sample after TAG hydrolysis. The HPLC–MS analyses of digestion products revealed that oxidized triacylglycerols are hydrolyzed by the digestive enzymes in a manner similar to that of native, unoxidized molecules. Significant amounts of sn-1(3)-MAG were found in all the samples after lipolysis, however, more of these were found in unoxidized rapeseed oil samples than in the oxidized oils. Several oxidized molecules were identified with the aid of synthesized oxylipids. This novel method is scalable to small-scale preparative fractionation of oxidized lipid molecules from a complex digestion sample. Also, the fingerprint-like, diagnostic, MS profiles of oxidized oils, reference compounds, and digestion products may be a great aid in comprehensive analysis of lipid oxidation and lipolysis.     

Separation of Sunflower Oil from Hexane by Use of Composite Polymeric Membranes

Vegetable oil extraction, as performed today by the oilseed-crushing industry, usually involves solvent extraction with commercial hexane. After this step, the vegetable oil–hexane mixture (miscella) must be treated to separate its components by distillation. If solvent-resistant membranes with good permeation properties can be obtained, membrane separation may replace, or be used in combination with, conventional evaporation. Two tailor-made flat composite membranes, poly(vinylidene fluoride) (PVDF–Si and PVDF–CA) and a commercially available composite membrane (MPF-50), were used to separate a crude sunflower oil–hexane mixture. The effects of temperature, cross-flow velocity (v), transmembrane pressure (Δp), and feed oil concentration (C _f) on membrane selectivity and permeation flux were determined. The PVDF–Si membrane achieved the best results, being stable in commercial hexane and having promising permselectivity properties for separation of vegetable oil–hexane miscella. Improved separation performance was obtained at C _f = 25%, Δp = 7.8 bar, T = 30 °C, and v = 0.8 m s⁻¹; a limiting permeate flux of 12 Lm⁻² h⁻¹ and 46.2% oil retention were achieved. Low membrane fouling was observed under all the experimental conditions studied.     

Free Fatty Acids in Commercial Krill Oils: Concentrations,Compositions, and Implications for Oxidative Stability

The concentrations and pro-oxidative effects of free fatty acids in commercial krill oil are not well defined. We now report that krill oil free fatty acids account for 2–13% of total lipids in commercial krill oil (n = 8) that these compounds are enriched in eicosapentaenoic acid (+7.1%) and docosahexaenoic acid (+6.3%) relative to whole oils; and that this composition make them highly pro-oxidizing in marine triacylglycerol oils, but not in krill oil, which derives oxidative stability from both its phospholipids, and neutral lipids (the latter because of astaxanthin). Specific fatty acid esterification patterns showed that krill oil free fatty acids predominantly (88–93%) originated from phospholipids, mainly from the sn-2 position, which was eight-fold more hydrolyzed than the sn-1 position. Lipolysis was not ongoing in stored oils. Adding small amounts of krill oil (1–5%) to marine triacylglycerol oils significantly increased their oxidative stability and also their resistance to free fatty acid-mediated pro-oxidative effects.     

Direct sampling of orujo oil for determining residual hexane by using a chemsensor

Hexane is used to extract edible oils from oleaginous seeds. The detection of hexane in orujo oil is mandatory, as its presence in the final product may negatively affect human health. Headspace-GC is the technique of choice for determining residual solvent in foods. In the present work, a new instrument based on the headspace principle and mass spectrometric detection without chromatographic separation, ChemSensor, is proposed for the direct screening of orujo oil to determine residual hexane. This instrument provided an overall response, corresponding to the volatiles profile, including that of hexane, which could not be directly discriminated. By selecting the m/z values corresponding to n-hexane (major component of commercial hexane), the selectivity of the method was good enough to determine residual hexane in the range of 2.0–65 μg mL⁻¹ (corresponding to 2.3–75.6 mg of hexane per kg of oil) with high precision. The detection limit achieved (0.7 mg per kg of oil) was lower than the maximum residual limit established by the European Union (5 mg per kg of oil). Two multivariate techniques, partial least squares and principal components regression (PCR), were compared with univariate regression; PCR provided the best results.     

Increase in the γ-linolenic acid content by solvent winterization of fungal oil extracted fromMortierella genus

The fungal oil extracted fromMortierella ramanniana var.angulispora (IFO 8187) was solvent winterized in order to raise the content of γ-linolenic acid (GLA). Effects of winterization conditions (solvent, oil concentration in the solvent and temperature) and changes of glyceride compositions were discussed. The fungal oil was separated into four diglycerides and 17 triglycerides (TG) with high performance liquid chromatography. The predominant species were POO, POP and LOP, whose contents were 24.4, 22.9 and 9.4% of the total TG, respectively. Ethanol at 4°C gave the highest GLA content of 10.5% in spite of lower yield than with acetone at −20°C. The highest separation efficiency for GLA (η_GLA) was 0.27 with acetone at −20°C and 10% oil concentration, resulting in 8.3% of GLA from the fungal oil at 5.7% LGA. In case of lower oil concentration at 5–20%, η_GLA showed higher in the following order: acetone (−20°C)>n-hexane (−20°C)>acetone (4°C)>petroleum ether (−20°C). The winterization process also proved to be effective for the separation of TG type, Sa₂U (Sa; saturated fatty acid; U, unsaturated fatty acid) into the crystallized fraction and SaU₂ into the liquid fraction. Acetone at −20°C showed higher separation efficiency for triunsaturated TG than the other solvents.     

Solvent extraction of the oils of rubber,melon, pumpkin and oilbean seeds

Solvents of differing dielectric constant were used to extract oils from the seeds of: rubber [Hevea brasiliensis (Kunth) Muell. Arg.], melon [Colocynthis vulgaris Schrad], fluted pumpkin [Telfairia occidentalis Hook f.] and oilbean [Pentaclethra macrophylla Benth]. The aim was to examine the effect of solvent polarity on oil yield and oil properties. The oils were extracted under Soxhlet conditions with the following solvents: petroleum benzene (60–80°C), cyclohexane, isopropyl ether, ethyl acetate, tetrahydrofuran, propan-2-ol and acetone. The oils were characterized by acid number, iodine value and color intensity determinations. The oil yields of each seed in different solvents ranged as follows: 58.0–64.4% (pumpkin), 56.1–59.1% (melon), 40.6–48.8% (rubber) and 35.4–43.3% (oilbean). The equilibrium extracting capacity of each solvent was found to depend on two factors, namely, the nature of the oil and the polarity of the solvent. Both factors were found to determine the acid number, iodine value and color intensity of each oil.     

The effect of relative concentrations on the efficiency of separation of polar and nonpolar lipids by alumina column chromatography

Columns were packed with aluminas from various sources and of low and high adsorptive power. Polar and nonpolar lipids were eluted from them with strong and weak solvents, and the fractions were tested for phosphorus, nitrogen, sterols, and triglycerides. The conditions used by the 1956 Technical Committee of the N.S.P.A. for the chromatography of linseed oil gave the most complete separation of polar from nonpolar lipids when the former were present in low concentration. When the lipid phosphorus load approached 0.4 mg./gm. alumina however, none of the alumina systems tried was satisfactory although silicic acid columns resolved the mixtures completely. Gamma alumina had greater resolving power thanalpha alumina monohydrate. Brockmann Grade I alumina with a strong solvent system gave better speration than weaker aluminas with weaker solvent systems. Presented at the fall meeting, American Oil Chemists' Society, Los Angeles, Calif., September 28–30, 1959. Contribution No. 44 from the Genetics and Plant Breeding Research Institute, Research Branch, Canada Department of Agriculture, Ottawa, Ontario.     

Selectivity in interpolymer complex formation between phenolic copolymer, polyelectrolyte, non-ionic polymer and metal ions

Summary Multicomponent complexes of the phenolic copolymer with specific coordinating groups with polyethylene imine (PEI), polyvinyl pyrrolidone (PVP), Cu²⁺ and Ni²⁺ ions has been studied in dimethyl formamide – methanol solvent mixture by several experimental techniques e.g. by viscometry, potentiometry, conductometry, IR and UV spectrophotometry and selective complexation, and mutual compatibility of the complementary polymers attached to the phenolic copolymer chain have been studied.     

Elucidation of Phosphatidylcholine Composition in Krill Oil Extracted from <Emphasis Type="Italic">Euphausia superba</Emphasis>

High performance liquid chromatography-electrospray tandem mass spectrometry was used to elucidate the phospholipids in krill oil extracted from Euphausia superba, an emerging source for human nutritional supplements. The study was carried out in order to map the species of the choline-containing phospholipid classes: phosphatidylcholine and lyso-phosphatidylcholine. In addition, the prevalent phosphatidylcholine class was quantified and the results compared with prior analysis. The qualification was performed with separation on a reverse phase chromatography column, while the quantification was obtained with class separation on a normal phase chromatography column. An Orbitrap system was used for the detection, and pulsed-Q dissociation fragmentation was utilized for the identification of the species. An asymmetrical exclusion list was applied for detection of phospholipid species of lower concentration, significantly improving the number of species observed. A total of 69 choline-containing phospholipids were detected, whereof 60 phosphatidylcholine substances, among others seven with probable omega-3 fatty acids in both sn-1 and sn-2. The phosphatidylcholine concentration was estimated to be 34 ± 5 g/100 g oil (n = 5). These results confirm the complexity of the phospholipid composition of krill oil, and the presence of long chained, heavily unsaturated fatty acids.     

Variation in fatty acid composition of the different acyl lipids in seed oils from fourSesamum species

Seeds from different collections of cultivatedSesamum indicum Linn. and three related wild species [specifically,S. alatum Thonn.,S. radiatum Schum and Thonn. andS. angustifolium (Oliv.) Engl.] were studied for their oil content and fatty acid composition of the total lipids. The wild seeds contained less oil (ca. 30%) than the cultivated seeds (ca. 50%). Lipids from all four species were comparable in their total fatty acid composition, with palmitic (8.2–12.7%), stearic (5.6–9.1%), oleic (33.4–46.9%) and linoleic acid (33.2–48.4%) as the major acids. The total lipids from selected samples were fractionated by thin-layer chromatography into five fractions: triacylglycerols (TAG; 80.3–88.9%), diacylglycerols (DAG; 6.5–10.4%), free fatty acids (FFA; 1.2–5.1%), polar lipids (PL; 2.3–3.5%) and steryl esters (SE; 0.3–0.6%). Compared to the TAG, the four other fractions (viz, DAG, FFA, PL and SE) were generally characterized by higher percentages of saturated acids, notably palmitic and stearic acids, and lower percentages of linoleic and oleic acids in all species. Slightly higher percentages of long-chain fatty acids (20∶0, 20∶1, 22∶0 and 24∶0) were observed for lipid classes other than TAG in all four species. Based on the fatty acid composition of the total lipids and of the different acyl lipid classes, it seems thatS. radiatum andS. angustifolium are more related to each other than they are to the other two species.     

Effect of absorbent in solid‐phase extraction on quantification of phospholipids in palm‐pressed fiber

Palm‐pressed fiber (PPF) is a by‐product of palm oil milling and it has been found to contain a high percentage of phospholipids (PL). The aim of this work was to evaluate the best solid‐phase extraction (SPE) method to purify PL from PPF. The purified PL were analyzed using high‐performance liquid chromatography (HPLC) with an evaporative light‐scattering detector (ELSD). The oil was extracted from PPF using the Folch method and classes of PL were purified using SPE. Different solvent phases and normal‐phase SPE cartridges [silica (Si), aminopropyl‐bonded silica (NH₂) and diol‐bonded silica (2OH)] at the same ratio of oil to sorbent mass were used to study the separation of PL from the extracted oil. The recovery of each standard PL was performed in a model oil system with the same amount of standard PL being added, and the repeatability of the SPE method was investigated. The isolation of PL by SPE diol cartridge and subsequent analysis by HPLC/ELSD have shown to be an accurate and reproducible analytical method for the determination of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and lysophosphatidylcholine in PPF. This method provided a high yield and rapid separation of PL in PPF with less contamination from other lipid groups.     

Thermophilic fungi: IV. The lipid composition of six species

The lipid composition of six thermophilic fungi (Myriococcum albomyces, Mucor miehei, Papulaspora thermophila, Rhizopus sp.,Thielavia thermophila (+)Thielavia thermophila (−), andTorula thermophila) was examined. The relative per cent total lipids (4.9–26.3%), neutral lipids (55.5–88.3%), polar lipids (11.7–44.6%) and the fatty acid profile of each lipid fraction was determined. The predominant fatty acids were 16∶0, 18∶0 and 18∶2, and lesser amounts of 12∶0, 14∶0, 15∶0, 16∶1, 16∶2, 17∶0 and 18∶3 were present. The total lipids contained an average of 0.96 double bonds per mole fatty acid (unsaturation index [USI]) the neutral lipids 0.86 USI and the polar lipids 0.84 USI, excluding the values forTorula thermophila. These data show a high degree of saturation and are consistent with data reported for other fungal thermophiles.Torula thermophila possessed abnormally high USI values (1.15–1.50) and was cultured at three different temperatures (25, 45 and 51 C). As the culture temperature ofTorula thermophila increased, the USI decreased. The USI of the polar lipids ofTorula thermophila at 25, 45 and 51 C were 1.50, 1.28 and 1.11, respectively. Thus the membrane lipids of this fungus appear unusual for a thermophile.     

Supercritical fluid extraction of fungal oil using CO2, N2O,CHF3 and SF6

The extraction of oil from fungi (Mortierella ramanniana var.angulispora) was studied using carbon dioxide (CO₂), nitrous oxide (N₂O), trifluoromethane (CHF₃) and sulfur hexafluoride (SF₆) under supercritical conditions. The oil solubility was highest in SC-N₂O followed by SC-CO₂, while both SC-CHF₃ and SC-SF₆ showed poorer solvent power. The recorded oil solubilities at 333 K and 24.5 MPa were 2.3 wt% in N₂O, 0.48 wt% in CO₂, 0.0099 wt% in CHF₃ and 0.0012 wt% in SF₆. The oil solubilities in SC-N₂O and SC-CO₂ were measured over the pressure range 15.7–29.4 MPa and at temperatures ranging from 313–353 K. N₂O always showed greater solvent power than did CO₂ at the same temperature and pressure. The solvent power of a supercritical fluid increases with density at a given temperature, and increases with temperature at constant density. The change in neutral lipid composition of the extracted oil with the extraction ratio was measured. Free fatty acids or diglycerides were extracted more easily than triglycerides or sterol esters. The change in fatty acid composition was also measured. The proportion of γ-linolenic acid in the extract remained constant throughout the extraction.     

Characterization of Turkish Virgin Olive Oils Produced from Early Harvest Olives

Olives were collected from various districts of Turkey (North and South Aegean sub-region, Bursa-Akhisar, South East Anatolia region) harvested over seven (2001–2007) seasons. The aim of this study was to characterize the chemical profiles of the oils derived from single variety Turkish olives including Ayvalik, Memecik, Gemlik, Erkence, Nizip Yaglik and Uslu. The olive oils were extracted by super press and three phase centrifugation from early harvest olives. Chosen quality indices included free fatty acid content (FFA), peroxide value (PV) and spectrophotometric characteristics in the ultraviolet (UV) region. According to the FFA results, 46% (11 out of 24 samples) were classified as extra virgin olive oils; whereas using the results of PV and UV, over 83% (over 19 of the 24 samples) had the extra virgin olive oil classification. Other measured parameters included oil stability (oxidative stability, chlorophyll pigment, pheophytin-α), cis–trans fatty acid composition and color index. Oxidative stability among oils differed whereas the cis–trans fatty acid values were within the national and international averages. Through the application of two multivariate statistical methods, Principal component and hierarchical analyses, early harvest virgin olive oil samples were classified according to the geographical locations categorized in terms of fatty acid profiles. Such statistical clustering gave rise to defined groups. These data provide evidence of the variation in virgin olive oil quality, especially early harvest and cis–trans isomers of fatty acid profiles from the diverse agronomic conditions in the olive growing regions of Turkey.     

Lipid,sterol and fatty acid composition of antarctic krill (Euphausia superba Dana)

The lipid classes, fatty acids of total and individual lipids and sterols of Antarctic krill (Euphausia superba Dana) from two areas of the Antarctic Ocean were analyzed by thin layer chromatography (TLC), gas liquid chromatography (GLC) and gas liquid chromatography/mass spectrometry (GLC/MS). Basic differences in the lipid composition of krill from the Scotia Sea (caught in Dec. 1977) and krill from the Gerlache Strait (caught in Mar. 1981) were not observed. The main lipid classes found were: phosphatidylcholine (PC) (33–36%), phosphatidylethanolamine (PE) (5–6%), triacylglycerol (TG) (33–40%), free fatty acids (FFA) (8–16%) and sterols (1.4–1.7%). Wax esters and sterol esters were present only in traces. More than 50 fatty acids could be identified using GLC/MS, the major ones being 14∶0, 16∶0, 16∶1(n−7), 18∶1(n−9), 18∶1(n−7), 20∶5(n−3) and 22∶6(n−3). Phytanic acid was found in a concentration of 3% of total fatty acids. Short, medium-chain and hydroxy fatty acids (C≤10) were not detectable. The sterol fraction consisted of cholesterol, desmosterol and 22-dehydrocholesterol.     

Metabolic Effects of Krill Oil are Essentially Similar to Those of Fish Oil but at Lower Dose of EPA and DHA,in Healthy Volunteers

The purpose of the present study is to investigate the effects of krill oil and fish oil on serum lipids and markers of oxidative stress and inflammation and to evaluate if different molecular forms, triacylglycerol and phospholipids, of omega-3 polyunsaturated fatty acids (PUFAs) influence the plasma level of EPA and DHA differently. One hundred thirteen subjects with normal or slightly elevated total blood cholesterol and/or triglyceride levels were randomized into three groups and given either six capsules of krill oil (N = 36; 3.0 g/day, EPA + DHA = 543 mg) or three capsules of fish oil (N = 40; 1.8 g/day, EPA + DHA = 864 mg) daily for 7 weeks. A third group did not receive any supplementation and served as controls (N = 37). A significant increase in plasma EPA, DHA, and DPA was observed in the subjects supplemented with n-3 PUFAs as compared with the controls, but there were no significant differences in the changes in any of the n-3 PUFAs between the fish oil and the krill oil groups. No statistically significant differences in changes in any of the serum lipids or the markers of oxidative stress and inflammation between the study groups were observed. Krill oil and fish oil thus represent comparable dietary sources of n-3 PUFAs, even if the EPA + DHA dose in the krill oil was 62.8% of that in the fish oil.     

Lipid composition of perilla seed

The composition of lipids and oil characteristics from perilla [Perilla frutescens (L.) Britt.] seed cultivars are reported. Total lipid contents of the five perilla seed cultivars ranged from 38.6 to 47.8% on a dry weight basis. The lipids consisted of 91.2–93.9% neutral lipids, 3.9–5.8% glycolipids and 2.0–3.0% phospholipids. Neutral lipids consisted mostly of triacylglycerols (88.1–91.0%) and small amounts of sterol esters, hydrocarbons, free fatty acids, free sterols and partial glycerides. Among the glycolipids, esterified sterylglycoside (48.9–53.2%) and sterylglycoside (22.1–25.4%) were the most abundant, while monogalactosyldiacylglycerol and digalactosyldiacylglycerol were present as minor components. Of the phospholipids, phosphatidylethanolamine (50.4–57.1%) and phosphatidylcholines (17.6–20.6%) were the major components, and phosphatidic acid, lysophosphatidylcholine, phosphatidylserine and phosphatidylinositol were present in small quantities. The major fatty acids of the perilla oil were linolenic (61.1–64.0%), linoleic (14.3–17.0%) and oleic acids (13.2–14.9%). Some of the physicochemical characteristics and the tocopherol composition of perilla oil were determined.     

Argentation supercritical fluid chromatography for quantitative analysis of triacylglycerols

A method for quantitative analysis of neutral lipids has been developed. Four different techniques have been combined for this purpose—supercritical fluid chromatography (SFC), silver ion chromatography, packed microcolumns and miniaturized evaporative light-scattering detection (ELSD). The development and optimization of the method are discussed. The separation of a series of vegetable, fish and hydrogenated oils was demonstrated. Application of eluent composition programming resulted in excellent separation of complex samples. Packed microcolumn argentation SFC provides at least as high a separation power as corresponding high-performance liquid chromatography methods. The combination of packed microcolumn SFC and miniaturized ELSD constitutes a powerful analytical system for the quantitative analysis of triacylglycerols.     

The unusual occurrence of 14-methylhexadecanoic acid in pinaceae seed oils among plants

14-Methylhexadecanoic (14-MHD) acid has been identified in a sample of pine seed oil (Pinus contorta) by gas-liquid chromatography-mass spectrometry of its picolinyl ester derivative: Its identification (through its equivalent chain length) and its distribution in four conifer families have been checked. It occurred only in Pinaceae, where it was found in 72 species belonging to the genera Pinus, Abies, Cedrus, Tsuga, Pseudotsuga, Larix, and Picea, in the range 0.02–1.15%. 14-MHD acid could not be detected in the lipids of Taxaceae (Taxus baccata), Cupressaceae (Juniperus communis), or Taxodiaceae (Sciadopytis verticillata), even after a 10-fold concentration of the saturated acid fraction isolated by argentation thinlayer chromatography. It is concluded that Pinaceae, along with Ginkgo biloba seed lipids, are major exceptions in the plant kingdom with regard to 14-MHD acid, which otherwise occurs almost exclusively in lipids of animals and microorganisms. The biosynthesis and metabolic role of 14-MHD acid, which other-wise also occur in wood and leaf lipids, remain unknown.