Free Sterol Composition Research Articles

Characterization of Seed Oil from Six In Situ Collected Wild Amaranthus Species

Six Amaranthus species (A. cruentus, A. hybridus, A. hypochondriacus, A. muricatus, A. tuberculatus, and A. viridis) were collected in Italy (wild habitats) from crops and roadsides. Amaranth seed oil was extracted to obtain fractions rich in squalene. Squalene, free fatty acid, tocopherol, and sterol composition and content were investigated in detail. An analysis of variance and principal components was performed. The oil content in the seed ranged from 5.17% (A. muricatus) to 12.20% (A. tuberculatus). The quantity of squalene in the oil varied from 3.43% (A. muricatus) to 6.09% (A. hypochondriacus). The primary sterols were beta-sitosterol, brassicasterol, campesterol, and stigmasterol. The main tocopherols in all the samples were alfa-tocopherol, beta-tocopherol, and delta-tocopherol. Our results exhibited that the smallest seeds (A. tuberculatus) have the highest percentages of oil and squalene, whereas the largest seeds size (A. muricatus) show the lowest percentages. There is also evidence that the samples growing at lower altitudes show the highest concentration of fatty acids. According to our results, the six wild Amaranthus species exhibited similar characteristics to commercial species. This study confirms that the site of the collection has an impact on the oil and squalene content of the Amaranthus species.

Heat stress leads to rapid lipid remodeling and transcriptional adaptations in Nicotiana tabacum pollen tubes.

After reaching the stigma, pollen grains germinate and form a pollen tube that transports the sperm cells to the ovule. Due to selection pressure between pollen tubes, pollen grains likely evolved mechanisms to quickly adapt to temperature changes to sustain elongation at the highest possible rate. We investigated these adaptions in tobacco (Nicotiana tabacum) pollen tubes grown in vitro under 22°C and 37°C by a multi-omics approach including lipidomic, metabolomic, and transcriptomic analysis. Both glycerophospholipids and galactoglycerolipids increased in saturated acyl chains under heat stress (HS), while triacylglycerols (TGs) changed less in respect to desaturation but increased in abundance. Free sterol composition was altered, and sterol ester levels decreased. The levels of sterylglycosides and several sphingolipid classes and species were augmented. Most amino acid levels increased during HS, including the noncodogenic amino acids γ-amino butyrate and pipecolate. Furthermore, the sugars sedoheptulose and sucrose showed higher levels. Also, the transcriptome underwent pronounced changes with 1,570 of 24,013 genes being differentially upregulated and 813 being downregulated. Transcripts coding for heat shock proteins and many transcriptional regulators were most strongly upregulated but also transcripts that have so far not been linked to HS. Transcripts involved in TG synthesis increased, while the modulation of acyl chain desaturation seemed not to be transcriptionally controlled, indicating other means of regulation. In conclusion, we show that tobacco pollen tubes are able to rapidly remodel their lipidome under HS likely by post-transcriptional and/or post-translational regulation.

Analysis of Free and Esterified Sterol Content and Composition in Seeds Using GC and ESI-MS/MS.

Total sterol content and composition in plant tissues can be easily determined by gas chromatography (GC) after saponification of the total lipid extract. However, in oleogenic tissues a significant proportion of the sterol is esterified to fatty acids, with GC methodologies unable to provide information about the proportion and the molecular species composition of intact steryl esters (SEs). Here we describe an electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and Multiple Reaction Monitoring (MRM) method which, in parallel with GC analysis, allows for the accurate determination of both free and esterified sterol content and composition in seeds. After extraction of seed oil with hexane, free sterols are derivatized with undecanoyl chloride, total steryl esters are then purified from triacylglycerol (TAG) by liquid chromatography, infused and ionized as ammonium adducts, with molecular species identified and quantified by fragmentation in the presence of internal standards.

Characterization and determination of free phytosterols and phytosterol conjugates: The potential phytochemicals to classify different rice bran oil and rice bran

Phytosterols are important beneficial compounds found in rice bran (RB) and rice bran oil (RBO). Although relationships have been confirmed between the forms of phytosterols and their bioactivities, the analysis of different forms of phytosterols in RB and RBO has been lacking. In this study, high temperature gas chromatography-mass spectrometry (HTGC-MS) was combined with the single standard to determine multi-components (SSDMC) method to determine free sterols (FSs) and steryl glycosides (SGs) in RB and RBO. High-performance liquid chromatography (HPLC) was used to determine steryl ferulates (SFs). There was clear variation in the composition of FS, SF and SG, indicating that different forms of phytosterols can discriminate between different RB and RBO. The developed method may be also useful for the detection of other compounds of interest in oils, oil seeds or cereals.

Nutrient quality of vertebrate dung as a diet for dung beetles

At the basis of a trophic web, coprophagous animals like dung beetles (Scarabaeoidea) utilize resources that may have advantages (easy gain and handling) as well as drawbacks (formerly processed food). Several studies have characterized the nutrients, e.g. C/N ratios and organic matter content, for specific types of dung. However, a comparative approach across dung types and feeding guilds of dung producers, and relationships between dung nutrients and preferences by coprophages, have been missing. Hence, we analyzed water content, C/N ratio, amino acid, neutral lipid fatty acid, free fatty acid and sterol composition and concentrations in dung from 23 vertebrates, including carnivore, omnivore and herbivore species. Our analyses revealed significant differences among the three vertebrate feeding guilds for most nutritional parameters. Although formerly processed, dung grants sufficient amounts of essential nutrients for insects. We tested whether nutrients can explain the dung beetles’ preferences in a field experiment, using 12 representative dung types in baits that were installed in 27 forests and 27 grasslands. Although consistent preferences for specific dung types were pronounced, the nutritional composition did not predict the variation in attractiveness of these dung diets, suggesting a primary role of dung volatiles irrespective of food quality.

The Content and Composition of Total, Free, and Esterified Sterols of Lotus Plumule Oil by GC–MS/FID

AbstractThis study investigated the content and composition of total, free, and esterified sterols of three varieties of lotus plumule oil (Hunan lotus, Jiangxi lotus, and Fujian lotus) using GC–MS/FID. The fatty acid composition of sterol fatty acid esters (SFAE) was also analyzed and compared with that of triglycerides. Results showed that total sterol of lotus plumule oil (12.10–14.21 g/100 g) was higher than that of other plant oils (corn germ oil, 1.11 g/100 g; rapeseed oil, 0.78 g/100 g). No significant difference was found among the total sterol contents of the three types of lotus plumule oils (p > 0.05). Most sterol existed in ester forms (81.8–89.1%) rather than in free forms (8.4–10.1%). β‐Sitosterol (71.4–73.4%), and campesterol (6.2–7.5%) were the predominant fractions of free sterols. β‐Sitosterol (41.3–53.7%) and ∆5‐avenasterol (27.1–31.1%) were the predominant fractions of esterified sterols, followed by campesterol (12.1–13.0%) and ∆7‐avenasterol (3.4–3.7%). Linoleic acid (63.6–65.8%), oleic acid (8.3–10.4%), and behenic acid (9.0–9.9%) were the main fatty acids of SFAE, which were different from those of triglycerides. The results from this study suggest that lotus plumule oil may be a good resource of SFAE and can be used as a supplemental ingredient in functional foods.

Designing of high voltage electric field for soybean and sunflower oil bleaching

Abstract To the best of our knowledge, this work for the first time in the world carried out at the Tarbiat Modares University. In the present research, high voltage electric field (HVEF) method was utilized for bleaching of soybean and sunflower oil. For optimization of the bleaching process, the effect of different independent variables such as clay percentage (0.5–2%), voltage (0–24 kV), distance (8–12 mm), different numbers of cathode–anode (4–8) and inert-electrolyte (HCl; 0–50 mM) was studied. In order to compare the properties of the bleached samples using HVEF and control (industrial condition) bleaching methods, the indexes of Lovibond, Hunter lab, carotenoid, chlorophyll, bleaching efficiency (BE), peroxide (PV), tiobarbituric acid (TBA), free fatty acid (FFA), tocopherol, sterol and fatty acid composition were measured. According to the results, the BE value of HVEF technique in bleached soybean (88.35%) and sunflower oil (75.41%) were higher in comparison with the soybean (85.80%) and sunflower (58.14%) oils bleached using control method. Consequently, the HVEF bleaching method seems to be an alternative to the industrial method as it reduced clay percentage (50 and 66%), temperature (28% and 28%) and process time (35% and 35%) in soybean and sunflower oil bleaching, respectively. Industrial relevance High voltage electric field (HVEF) method for bleaching of soybean and sunflower oil was done, for the first time in the world to the based of our knowledge, and was optimized under different conditions including clay percentage (0.5–2%), voltage (0–24 kV), distance (8–12 mm) with different numbers of cathode–anode (0–8), inert electrolyte (HCl; 0–50 mM). HVEF could be an alternative for bleaching method in the oil industry; HVEF could reduce %clay, temperature and time consumption in oil industry with low energy consumption; HVEF could produce micro and Nano clay by pulsed corona and pulsed spark type systems. -To Patent No: A23D ; C11B ; C11C ; H03 , “Design of high voltage electric field for the oil bleaching”, Tehran-Iran (2015).

Plant Sterol Diversity in Pollen from Angiosperms.

Here we have examined the composition of free sterols and steryl esters of pollen from selected angiosperm species, as a first step towards a comprehensive analysis of sterol biogenesis in the male gametophyte. We detected four major sterol structural groups: cycloartenol derivatives bearing a 9β,19-cyclopropyl group, sterols with a double bond at C-7(8), sterols with a double bond at C-5(6), and stanols. All these groups were unequally distributed among species. However, the distribution of sterols as free sterols or as steryl esters in pollen grains indicated that free sterols were mostly Δ(5)-sterols and that steryl esters were predominantly 9β,19-cyclopropyl sterols. In order to link the sterol composition of a pollen grain at anthesis with the requirement for membrane lipid constituents of the pollen tube, we germinated pollen grains from Nicotiana tabacum, a model plant in reproductive biology. In the presence of radiolabelled mevalonic acid and in a time course series of measurements, we showed that cycloeucalenol was identified as the major neosynthesized sterol. Furthermore, the inhibition of cycloeucalenol neosynthesis by squalestatin was in full agreement with a de novo biogenesis and an apparent truncated pathway in the pollen tube.

Characterisation of the geographical origin of Western Greek virgin olive oils based on instrumental and multivariate statistical analysis

In this work, measurements of free acidity, peroxide content, spectrophotometric parameters, chlorophyll content, phenol, sterol, fatty acid and triacylglycerol composition, were carried out on samples of virgin olive oils (VOOs) coming from four different Greek Ionian islands, i.e. Zakynthos, Kefalonia, Lefkada and Kerkyra. An analysis of variance (ANOVA) highlighted statistically significant differences (p<0.01) in the values of 26 analytical parameters amongst the VOOs produced in the four different geographical regions but a post-hoc test showed that no variable was able to distinguish all four origins. Analogously, a Principal Component Analysis (PCA) showed a modest grouping of VOOs according to geographical origin except for Kerkyra samples which were more distinct from others. Applying discriminant function analysis (DFA) a good separation of the four geographical groups was achieved with classification and prediction abilities equal to 97.7% and 95.3%, respectively. Moreover, the analysis of the standardised coefficients showed that the fatty acids and triacylglycerols were the most discriminant variables. This last outcome was confirmed by comparison of the prediction performances obtained applying DFA on four subdatasets containing fatty acids (69.8%), triacylglycerols (76.7%), sterols (62.8%), and remaining parameters (65.1%) together, respectively. As the results showed, the multidisciplinary approach that combines different types of analytical determinations improved the discrimination of geographical origin for Greek virgin olive oils.

Free Sterol Composition of Species in the Dinoflagellate GenusPyrocystis: A Spectrum of Sterol Diversity

The dinoflagellate genus Pyrocystis includes a small number of marine species, which spend the majority of their life cycles as nonmotile cells within a carbohydrate sheath, and which are found ubiquitously throughout the world's oceans. The biochemistry of this model dinoflagellate genus has been widely studied due to its ability to bioluminesce. However, Pyrocystis has been comparatively understudied with respect to its lipid biochemistry, in particular that of sterols. To date, examination of the sterols of Pyrocystis has focused primarily upon Pyrocystis lunula, which produces cholesterol and 4,24-dimethyl-5α-cholestan-3β-ol as its predominant sterols, while it lacks the common dinoflagellate sterol, dinosterol. We have examined the sterol composition of the two other commercially available species of Pyrocystis, Pyrocystis fusiformis and Pyrocystis noctiluca. Pyrocystis noctiluca possesses dinosterol as its most abundant sterol, while P. fusiformis possesses dinosterol and 4,24-dimethyl-5α-cholestan-3β-ol as the predominant sterols, placing it at an intermediate position between P. lunula and P. noctiluca, as based on sterol composition. The potential limitations of the dinoflagellate sterol biomarker dinosterol are also explored in this study due to its notable absence in P. lunula.

Cholesterol Substitution Increases the Structural Heterogeneity of Caveolae

Caveolin-1 binds cholesterol and caveola formation involves caveolin-1 oligomerization and cholesterol association. The role of cholesterol in caveolae has so far been addressed by methods that compromise membrane integrity and abolish caveolar invaginations. To study the importance of sterol specificity for the structure and function of caveolae, we replaced cholesterol in mammalian cells with its immediate precursor desmosterol by inhibiting 24-dehydrocholesterol reductase. Desmosterol could substitute for cholesterol in maintaining cell growth, membrane integrity, and preserving caveolar invaginations. However, in desmosterol cells the affinity of caveolin-1 for sterol and the stability of caveolin oligomers were decreased. Moreover, caveolar invaginations became more heterogeneous in dimensions and in the number of caveolin-1 molecules per caveola. Despite the altered caveolar structure, caveolar ligand uptake was only moderately inhibited. We found that in desmosterol cells, Src kinase phosphorylated Cav1 at Tyr(14) more avidly than in cholesterol cells. Taken the role of Cav1 Tyr(14) phosphorylation in caveolar endocytosis, this may help to preserve caveolar uptake in desmosterol cells. We conclude that a sterol C24 double bond interferes with caveolin-sterol interaction and perturbs caveolar morphology but facilitates Cav1 Src phosphorylation and allows caveolar endocytosis. More generally, substitution of cholesterol by a structurally closely related sterol provides a method to selectively modify membrane protein-sterol affinity, structure and function of cholesterol-dependent domains without compromising membrane integrity.

Lipid composition of the pacific lion-paw scallop, Nodipecten subnodosus, in relation to gametogenesis : 2. Lipid classes and sterols

Knowledge on the energy distribution during the reproductive cycle of the giant lion-paw scallop, Nodipecten subnodosus is limited, but necessary for aquaculture purposes. In a previous work, we found that natural food availability can sustain the full cost of gametogenesis in optimal environmental conditions. The present study analyses the lipid and sterol classes’ composition of several tissues during gametogenesis. In female gonads, the quantity of triacylglycerol (TG) increased parallel to gametogenesis from December (spawned gonads, 0.7 mg g − 1 or 7% of total lipids) to June (ripe gonads, 52 mg g − 1 or 66% of total lipids). In male gonads sampled in June, the concentration of TG (0.7 mg g − 1 or 3.5% of total lipids) and esterified sterols was significantly lower than in female gonads. Total phospholipids (PL) increased two-fold in female gonads of scallops sampled in June compared to the other months, which could be a result of oocyte production and growth, but no differences were found in the concentration of PL or in free sterol composition between male and female gonads. TG concentration in the digestive gland was similar in December and February (37–38 mg g − 1 or 68% of total lipids), but increased eight-fold by April and remained high in June (252–277 mg g − 1 or 90% of total lipids). Together with the increase in esterified sterols, these data suggest an increase in food availability during these months. A lack of decrease of TG levels in digestive gland during gonad development is in accordance to sufficient food availability in this location to satisfy the energetic demands of gametogenesis. Cholesterol was the principal sterol found in all tissues; its concentration, as well as the concentration of several phytosterols, remained constant throughout the maturation process, except in digestive gland where they increased from December to June, further supporting an increased availability of food. However, the presence of several phytosterols in substantial concentrations, mainly brassicasterol and 24-methylenecholesterol in gonad and somatic tissues, further supports a specific physiological role of these sterols in pectinids.

Effects of temperature and nutrient availability on plasma membrane lipid composition in Scots pine roots during growth initiation.

We studied the effects of root zone temperature (RZT) and nutrient availability on free sterols and phospholipids in the plasma membrane (PM) and on PM-ATPase activity in roots of 1-year-old Scots pine (Pinus sylvestris L.) seedlings during growth initiation in the spring. Seedlings were grown for 6 weeks in hydroponic cultures with low (0.5 mM N; LN) or high (3 mM N; HN) nutrient availability. The root zone was subjected to slow warming (SW) and fast warming (FW) treatments while maintaining similar air temperatures in both treatments. Decreases in the amount of phospholipids and in the phospholipid/free sterol ratio, an increase in the degree of saturation of phospholipid fatty acids and changes in free sterol composition were observed during root growth initiation. Changes in lipid composition of the PM associated with the cold deacclimation process were detected at RZTs above 9 degrees C. Nutrient availability affected the lipid composition of the PM only when RZT was increased slowly. When RZT increased from 4 to 6 degrees C in the SW treatment, the degree of saturation of phospholipid fatty acids decreased, especially in HN seedlings. The sitosterol/stigmasterol ratio remained higher in HN seedlings than in LN seedlings. After an RZT of 9 degrees C had been reached in the SW treatment, HN caused increases in the saturation of phospholipid fatty acids and root PM-ATPase activity, and a decrease in the phospholipid/free sterol ratio. Possible effects of changes in PM lipid composition on root growth and PM-ATPase activity are discussed.

SUT1 suppresses sec14-1 through upregulation of CSR1 in Saccharomyces cerevisiae.

SUT1 constitutive expression in aerobiosis suppressed the ts phenotype of the sec14-1 mutation, restored growth of the sec14-null mutant and corrected the translocation defect of the vacuolar carboxypeptidase Y. Therefore SUT1 was shown to be a novel potent sec14-1 suppressor. Further, the hypoxic gene CSR1 (YLR380W), a Sec14 homolog, was upregulated upon SUT1 constitutive expression. In addition, SUT1 effects on both sec14-1 suppression and on free sterol composition were abolished in a csr1-null background, showing that this gene acts downstream of SUT1.

SUT1 suppresses sec14-1 through upregulation of CSR1 in Saccharomyces cerevisiae

SUT1 constitutive expression in aerobiosis suppressed the ts phenotype of the sec14-1 mutation, restored growth of the sec14-null mutant and corrected the translocation defect of the vacuolar carboxypeptidase Y. Therefore SUT1 was shown to be a novel potent sec14-1 suppressor. Further, the hypoxic gene CSR1 (YLR380W), a Sec14 homolog, was upregulated upon SUT1 constitutive expression. In addition, SUT1 effects on both sec14-1 suppression and on free sterol composition were abolished in a csr1-null background, showing that this gene acts downstream of SUT1.

Inherent and growth stage-related differences in growth and lipid and sterol composition of algal species sensitive and tolerant to sterol-inhibiting fungicides

Fourteen algal species representing five algal classes were evaluated as to the effects of four sterol-inhibiting fungicides (SIFs) on growth. Sensitivity to SIFs was related to species, chemical, and concentration. Species of Chlorella sensitive and tolerant to SIFs were further compared relative to inherent and growth stage-related differences in lipid, free sterol (FS), and steryl ester (SE) composition. Inherent and growth stage-related qualitative and quantitative differences in FS and SE composition were apparent between the two species examined. The qualitative FS and SE composition was particularly different. The importance of lipids, FS, and SE in membrane structure and control of membrane function is well established. Species differences and environmentally (growth stage, light, temperature, and nutrition) induced changes in lipid composition may be part of the basis for sensitive and tolerant responses to SIFs and xenobiotics in general. The ability of lipophilic xenobiotics to partition into membranes most likely depends on the qualitative and quantitative composition of membrane components, at any given time, and seems to be highly dynamic. Although the biochemical mechanism of action of SIFs is well established, the ability for passive bioaccumulation of such compounds in algae may be equally as important.

Free and Esterified Sterol Composition of Edible Oils and Fats

The free and esterified sterol concentrations in 31 edible oils and fats were determined, including individual values for sitosterol, campesterol, stigmasterol, brassicasterol, Δ5-avenasterol, sitostanol, campestanol, and cholesterol. Free and esterified sterols were separated by solid-phase extraction (SPE), saponified, and quantified as trimethylsilyl ether derivatives using capillary gas–liquid chromatography. Considerable variability in the proportion of free and esterified sterols was observed among different oils and fats, with free sterols ranging from 32 to 94% of total sterols. Refining or hydrogenation tended to decrease total sterols and increase esterified sterols. Differences in total phytosterol content and the proportion of free and esterified sterols also were evident for different samples of some oils. Among four soybean oils, total phytosterols ranged from 205 to 287 mg/100 g, and free sterols constituted 68–81% of total sterols; two different shortenings differed 62% in total sterol content (185–301 mg/100 g). Such variability in phytosterol composition likely reflects differences in processing, growing season, or variety of a particular plant source.

Changes in the composition of fatty acids and sterols of membrane lipids during induction and differentiation of Brassica napus (var. oleifera L.) callus

Changes in the membrane lipid and sterols content and composition were studied during induction and differentiation in callus cultures of Brassica napus var. oleifera. Callus induction was associated with an increase of DGDG content, significant changes in fatty acids composition of all lipid fractions and increased degree of lipid unsaturation. The membrane lipid composition of tissue at different degrees of differentiation was found to vary significantly, particularly two weeks after transfer of callus to regeneration medium. The main differences concerned the content and composition of galactolipids. Curiously in many cases, these differences declined during subsequent culture, in spite of the morphogenesis process which was in progress. Another result of differentiation was the change in free sterol composition: in shoot regenerating calli the content of stigmasterol had rose whereas the accumulation of campesterol decreased. Even though observed changes in membrane properties may not play a role in morphogenesis they are nevertheless useful as developmental markers and can be invaluable in understanding biochemical basis of morphogenesis.

The sterols of Astragalus species from Iran: GLC separation and quantification

This paper presents the content and composition of free sterols in a number of Astragalus species from Iran. Free sterols of roots and seeds were analysed quantitatively and qualitatively by gas-liquid chromatography. Four major 3-beta-hydroxysterols (cholesterol, campesterol, stigmasterol and beta-sitosterol) were identified in roots of 18 species and in seeds of five species of Astragalus. beta-Sitosterol was the main free sterol in both roots and seeds. The cholesterol content in seeds was higher than that in roots. Trace amounts of cholesterol were detected in roots of only four species. beta-Sitosterol accounted for 34.4-69.2 and 76.8-81.8% of the total lipid content in roots and seeds, respectively. The beta-sitosterol content in roots and seeds of Astragalus species varied from 6.3 to 35.7 and 93.3 to 164.5mg/100g dry weight, respectively. Total free sterol content in seeds varied from 112.09 to 204.04mg/100g dry weight, significantly higher than those in roots (7.5 to 43.4mg/100g dry weight). The chemotaxonomic values of free sterols and total lipid profiles of some of the representative species are discussed.

Free and sulfated sterols of two far-east Leptasterias starfish

NMR spectroscopy, capillary GLC, and GLC-MS are used to study the composition of free and sulfated sterols from the far-east starfishLeptasterias alaskensis asiatica(Fischer) andL. Fisheri(Djakonov). The total free sterols of both species are shown to have similar qualitative and quantitative compositions and contain mainlyΔ7-sterols. Sterol sulfate fractions contain cholesterol sulfate as the main component but differ in the ratios ofΔ5:δ0:δ7-sterol derivatives. Possible reasons for these differences are discussed. A new steroid, 3β-hydroxycholest-5-en-7-one sulfate, was isolated.