Sterol concentration and distribution in sunflower seeds (Helianthus annuus L.) during seed development

Sunflower seeds are currently used for edible oil production. Among oil minor compounds, phytosterols are of special interest due to their cholesterol reducing properties. This paper reports studies on their accumulation and distribution in the embryo and hull, and the effects of temperature on phytosterol contents in sunflower seed produced under both conventional and organic field conditions. An optimised method of sterol determination, adapted to studies on small samples of seed, is presented. Seventy-two percent of phytosterols were found in the embryo, 28% in the hull. The periods of phytosterols concentration varied according to sterol category and seed part. Application of these results to improve production of natural sterols for functional food use is discussed.     

Characteristics and composition of Washingtonia filifera (Linden ex André) H. Wendl. seed and seed oil

Characteristics of seeds and oil extracted from Washingtonia filifera seeds are evaluated. The percentage composition of the W. filifera seeds is: ash 1.37%, oil 16.30%, protein content 3.46%, total carbohydrate 77.19% and moisture 3.22%. The major nutrients (mg/100 g of seeds) found in the seeds are: potassium (67.33), magnesium (34.35), calcium (187.85) and phosphorus (23.26). Physicochemical properties of the oil include: iodine value 67.33 g/100 g of oil; saponification value, 191.63 mg KOH/g of oil; refractive index (25 °C), 1.469; unsaponifiable matter, 0.83%; acidity, 0.41%; p-anisidine value, 0.87; peroxide value, 7.60 mEq O₂/kg of oil; carotenoid content 14.8 mg/100 g and the chlorophyll content = 0.13 mg/100 g. W. filifera seed oil shows some absorbance in the UV-B and UV-C ranges with potential use as a broad spectrum UV protectant. The oil contains high levels of oleic acid (40.60%) followed by lauric acid (17.87%), linoleic acid (16.26%), myristic acid (11.43%) and palmitic acid (9.23%). The triacylglycerols (TAGs) with equivalent carbon number ECN 44 (20.47%) are dominant, followed by TAGs ECN 46 (16.71%), TAGs ECN 42 (15.43%) and TAGs ECN 48 (15.41%). The DSC melting curves reveal that: melting point = 2.25 °C and melting enthalpy = 82.34 J/g. γ-Tocotrienol is the major tocol (72%) with the rest being δ-tocotrienol and α-tocotrienol. The results of the present analytical study show that W. filifera seed oil could be used in cosmetic, pharmaceutical and food products.     

Enzymatic browning and after-cooking darkening of Jerusalem artichoke tubers (Helianthus tuberosus L.)

Jerusalem artichoke tubers (Helianthus tuberosus L.) undergo enzymatic browning when peeled or cut, and turn grey after boiling, due to after-cooking darkening reactions between iron and phenolic acids. In an attempt to reveal the components responsible for these discolouration reactions, sensory evaluation and instrumental colour measurements were related to contents of total phenolics, phenolic acids, organic acids and iron in three varieties of raw and boiled Jerusalem artichoke tubers harvested in the autumn and the spring. No differences were found between varieties in sensory evaluated enzymatic browning, but Rema and Draga had higher scores than Mari in after-cooking darkening. Jerusalem artichoke tubers had higher contents of total phenolics, phenolic acids and citric acid in the autumn and low contents in the spring, while it was the opposite for malic acid. None of the chemical parameters investigated could explain the discolouration of the Jerusalem artichoke tubers.     

Identification and quantification of phenolic compounds from sunflower (Helianthus annuus L.) kernels and shells by HPLC-DAD/ESI-MS

Phenolic compounds were extracted from defatted sunflower (Helianthus annuus L.) kernels and shells and characterised by HPLC with diode array and electrospray ionisation (ESI) mass spectrometric detection in the negative mode. Quantification of individual compounds was carried out by external calibration. Among the eleven compounds analysed 5-O-caffeoylquinic acid was predominant amounting up to 59.1 mg/100 g in the shells and 3050.5 mg/100 g in the kernels. The specific fragmentation patterns of mono- and dihydroxycinnamoylquinic acids allowed the unambiguous distinction of several stereoisomers which have not been described for sunflower seeds and seed shells so far. The total phenolic content of about 4200 mg/100 g on a dry matter basis revealed defatted sunflower meal to be a promising source of phenolic compounds that might be recovered and used as natural antioxidants. Furthermore, the press residues originating from sunflower oil extraction were shown to be still rich in phenolic antioxidants, thus, providing the opportunity to valorize these by-products in terms of sustainable agricultural production.     

Characterisation of fatty acids and bioactive compounds of kachnar (Bauhinia purpurea L.) seed oil

Information concerning the exact composition of kachnar (Bauhinia purpurea) seed oil is scare. In the present contribution, a combination of CC, GC, TLC and normal-phase HPLC were performed to analyse lipid classes, fatty acids and fat-soluble bioactives of kachnar seed oil. n-Hexane extract of kachnar oilseeds was found to be 17.5%. The amount of neutral lipids in the crude seed oil was the highest (ca. 99% of total lipids), followed by glycolipids and phospholipids, respectively. Linoleic, followed by palmitic, oleic and stearic, were the major fatty acids in the crude seed oil and its lipid classes. The ratio of unsaturated fatty acids to saturated fatty acid, was higher in neutral lipid classes than in the polar lipid fractions. The oil was characterised by a relatively high amount of phytosterols, wherein the sterol markers were β-sitosterol and stigmasterol. β-Tocopherol was the major tocopherol isomer with the rest being δ-tocopherol. In consideration of potential utilisation, detailed knowledge of the composition of kachnar (B. purpurea) seed oil is of major importance.     

Effect of pectinase on the oil solvent extraction from different genotypes of sunflower (Helianthus annuus L.)

Pectinase-assisted solvent extraction of oil and total tocopherol from sunflower hybrids with different structural characteristics was studied by means of laboratory tests carried out in a stirred batch extractor at 50 °C. From the oil yield results obtained for the two hybrids studied, black-hull (P20) and striped-hull hybrids (ACA 884), it could be observed that oil was extracted at different rates in both hybrids. Analyzing the performance data versus time in the control samples, ACA 884 oil was released more easily and remained invariant with time, whereas for P20 the extraction was slower and the maximum value was reached at 60 min. The fiber wall structure collapsed by the enzymatic action, improving oil release and allowing a faster extraction. The efficiency of the treatment was measured in terms of the percentage of increase in oil yield compared with the control samples. It was observed that in hybrid ACA 884 the mean increases were not significantly different (p = 0.9295) over time (4.12% d.b.). Hybrid P20 showed a significant increase in the percentage of oil yield (p = 0.0001) at 10 min (10.85% d.b.), and then it remained virtually constant over time (4.89% d.b.). The black-hull hybrid (P20) presented significant differences in fatty acid composition due to the enzymatic action. Pectinase treatment was highly effective in the tocopherol extraction from hybrid P20 (p < 0.001), obtaining a 32.3% increase on average. The maximum values were 662 and 220 μg g dried ground seed⁻¹ at 60 and 120 min for black-hull and striped-hull hybrids, respectively. The enzymatic treatment effectively produced an increase in oil and tocopherol yields for the black-hull hybrid.     

Supercritical CO2 extraction optimization of pomegranate (Punica granatum L.) seed oil using response surface methodology

This study examined extraction of pomegranate seed oil by using supercritical carbon dioxide. Response surface methodology was used to evaluate the effects of the process parameters, namely extraction pressure, temperature and CO₂ flow rate on the yield of pomegranate seed oil. The extraction parameters were optimized with a central composite design experiment. The linear term of pressure, followed by the linear term of CO₂ flow rate, the quadratic terms of pressure, temperature and CO₂ flow rate and the interactions between pressure and temperature, as well as CO₂ flow rate and temperature, had significant effects on the oil yield (p < 0.05). Maximum yield of pomegranate seed oil from the mathematical model was predicted to be 156.3 g/kg dry basis under the condition of pressure 37.9 MPa and temperature 47.0 °C with CO₂ flow rate of 21.3 L/h. The fatty acid composition and the tocopherols' content of pomegranate seed oil extracted using supercritical CO₂ were compared with those obtained by Soxhlet method. Minor difference was found in the fatty acid composition of the oils extracted by the two methods. The content of total tocopherols was about 14% higher in the oil extracted with supercritical CO₂ than that obtained by Soxhlet extraction.     

Characterization of wax esters,free fatty alcohols and free fatty acids of crude wax from sunflower seed oil refineries

The sunflower seed wax (10–12%) from oil refineries was purified through two steps, namely, extraction using solvents and precipitation with chilled acetone. Fatty esters, free fatty alcohols and free fatty acids were separated by thin-layer chromatography (TLC). The fractions of fatty esters and free fatty alcohols were separated and identified by gas chromatography (GC) using a Dexil-300 column and OV-101 column, respectively, at temperatures from 150–290 °C and were further confirmed by Gas chromatography–mass spectrometry (GC–MS), using a capillary silica column SPB™-1 coated with polydimethyl siloxane. Among the long chain fatty esters (C₃₈–C₅₄), comprised of one unit of fatty alcohol and one unit of fatty acid through an ester linkage, the major esters were C₄₀–C_44, of which C₄₂ was predominant. These fatty esters, upon hydrolysis, followed by methylation and subsequent GC–MS analysis, showed the presence of C₁₆–C₃₀ fatty acids in the bound form, whereas methylation of native wax indicated the presence of C₁₈–C₃₀ fatty acids in the free form. Acetylation of hydrolysis product showed the presence of C₁₈–C₃₀ fatty alcohols in the bound form, where as acetylation of native wax indicated that the C₁₈–C₃₂ fatty alcohols were in the free form. n-Triacontanol (C₃₀), a plant growth regulator, was found in both free and bound forms.     

The in vitro and in vivo antioxidant properties of seabuckthorn (Hippophae rhamnoides L.) seed oil

The antioxidant capacity of seabuckthorn (Hippophae rhamnoides L.) seed oil was investigated with a number of established in vitro assays and in an in vivo study of carbon tetrachloride (CCl₄)-induced oxidative stress in mice. The results showed that DPPH radical scavenging activity, ferrous ion chelating activity, reducing power and inhibition of lipid peroxidation activity all increased with increasing concentrations of seabuckthorn seed oil. Moreover, the EC₅₀ values of seabuckthorn seed oil from the hydrogen peroxide, superoxide radical, hydroxyl radical scavenging assays were 2.63, 2.16 and 0.77 mg/ml, respectively. In the in vivo study, seabuckthorn seed oil inhibited the toxicity of CCl₄, as seen from the significantly increased activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The GSH content in the liver was also increased, whereas hepatic malondialdehyde was reduced. Taken together, these results clearly indicate that seabuckthorn seed oil has significant potential as a natural antioxidant agent.     

Sterol content in sunflower seeds (Helianthus annuus L.) as affected by genotypes and environmental conditions

Phytosterols play essential roles in many plant cell mechanisms. They are of industrial interest since, as part of the diet, they can reduce low density lipoprotein cholesterol. An increase in plant sterol contents, by improved crop varieties or crop management, could help to answer industrial demands and also to develop environmentally friendly extraction methods. The aim of this study was to evaluate genotypic variability of sterol content in cultivated sunflower and, in particular, effects of sowing date. Results showed large variability among a collection of sixteen sunflower inbred lines and hybrids. Total sterols varied almost twofold between extreme genotypes. A delay of sowing, giving higher temperatures during seed formation, induced a general increase in total sterol concentration by up to 35%, as well as variation in sterol composition according to genotype. These results are considered with an aim of improving sterol content by sunflower breeding programmes.     

Free radical scavenging activities and bioactive substances of Jerusalem artichoke (Helianthus tuberosus L.) leaves

The total phenolic content and radical scavenging activities of Jerusalem artichoke (Helianthus tuberosus L.) leaves were investigated. Results indicated that the ethyl acetate fraction contained the highest total phenolic content (266.69 ± 2.51 mg GAE/g dry extract) accompanied with strongest free radical scavenging abilities. Following an in vitro radical scavenging activity-guide fractionation procedure, six phenolic compounds which strongly quenched free radicals were separated from ethyl acetate fraction. Among them, 3-O-caffeoylquinic acid and 1,5-dicaffeoylquinic acid played a dominant role due to their strong free radical scavenging abilities and their high contents. The content of 3-O-caffeoylquinic acid in n-butanol fraction was 74.58 ± 1.05 mg/g, while 1,5-dicaffeoylquinic acid in ethyl acetate fraction was 104.51 ± 2.86 mg/g. The results imply that the leaves of Jerusalem artichoke might be a potential source of natural antioxidants.     

The most important bioactive components of cold pressed oil from different pumpkin (Cucurbita pepo L.) seeds

The composition and content of certain bioactive components of the cold pressed oil obtained from six samples of pumpkin seeds (Cucurbita pepo L.) cultivated in Serbia were analyzed by GC and GC/MS. The composition and content of fatty acids, tocopherols and phytosterols, and the total content of squalene were determined. The results indicate oil's excellent quality, with high contents of monounsaturated fatty acids (37.1 ± 0.70–43.6 ± 0.69 g/100 g of total fatty acids), total tocopherols (38.03 ± 0.25–64.11 ± 0.07 mg/100 g of oil), sterols (718.1 ± 6.1–897.8 ± 6.8 mg/100 g of oil) and especially squalene (583.2 ± 23.6–747 ± 16 mg/100 g of oil). High content of squalene, phytosterols and monounsaturated fatty acids recommend the use of this type of the oil in the nutritional and medical purposes.     

Fat and fatty acids of white lupin (Lupinus albus L.) in comparison to sesame (Sesamum indicum L.)

The study was undertaken to compare fat and fatty acid profiles in white lupin (Lupinus albus ssp. albus) and sesame (Sesamum indicum L.), representing two different families, Fabaceae and Pedaliaceae. Fat levels were 10.74% and 55.44% in seeds of white lupin and sesame, respectively. The results indicated that oleic, linolenic and arachidic acids in seed fat were higher in white lupin than in sesame cultivars. Oleic acid was the predominant fatty acid in white lupin, whereas linoleic acid was predominant in sesame. Fat content (%) was statistically significantly correlated with linoleic, linolenic and arachidic acids at the genotypic level. The fatty acid composition of white lupin is useful for human consumption. Although oil content of white lupin was lower than that of sesame, white sweet lupin could be improved.     

Inhibition of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) development with essential oil extracted from Cymbopogon schoenanthus L. Spreng. (Poaceae), and the wasp Dinarmus basalis (Rondani) (Hymenoptera: Pteromalidae)

The control of the development of Callosobruchus maculatus was studied using a method that combined exposure to essential oil extracted from Cymbopogon schoenanthus and the introduction of a pteromalid natural enemy of the bruchid, Dinarmus basalis. The effect of the essential oil used was evaluated on all developmental stages of C. maculatus and on adults of D. basalis. At the highest concentration tested (33.3 μl/l) all adults of C. maculatus were killed within 24 h of exposure to the oil and the development of newly laid eggs and neonate larvae was also inhibited. However, the oil had variable efficacy against the bruchid instars developing inside the seeds: 5-day-old larvae (63% LI and 37% LII) of C. maculatus developing inside the seeds proving to be highly susceptible while 15-day-old insects (84% of pupae and 16% of larvae) were tolerant. Under the same conditions (33.3 μl/l), adults of D. basalis were very susceptible to oil vapours and to the residual activity of the oil after 3 or 6 days. However, the introduction of 10 pairs of adult D. basalis into a jar containing 100 hosts aged 10 days, 3 or 6 days before the oil application, gave respectively an emergence of 26 or 18 adults of the parasitoid compared to 28 in the control and there was no adult emergence of the host. The possibility of an integrated pest management strategy by using allelochemicals such as essential oils and indigenous natural enemies to control C. maculatus development in cowpea stocks is discussed.     

Insecticidal activities of constituents identified in the essential oil from leaves of Chamaecyparis obtusa against Callosobruchus chinensis (L.) and Sitophilus oryzae (L.)

Analysis by gas chromatography-mass spectrometry led to the identification of 20 volatiles from the steam distilled oil of the leaves from Chamaecyparis obtusa Siebold et Zuccarni. Seven constituents of the oil were tested for contact and fumigant activity against adults of Callosobruchus chinensis (L.) and Sitophilus oryzae (L.). Responses varied with compound and dose rather than increasing exposure time beyond 1 or 2 days. In the impregnated-paper test with the constituents [bornyl acetate, (+)-limonene, myrcene, α-phellandrene, α-pinene, sabinene and terpinolene], at 0.1 mg/cm², bornyl acetate, α-phellandrene and terpinolene caused 97%, 97% and 87% mortality, respectively, against C. chinensis adults 1 day after treatment. At 0.05 mg/cm², moderate activity was achieved with bornyl acetate (56% mortality), α-phellandrene (75%) and terpinolene (55%). Against S. oryzae adults, at 0.26 mg/cm², terpinolene caused 93% mortality 2 days after treatment, whereas 80% mortality 4 days after treatment was observed with bornyl acetate and α-phellandrene. The toxicity of these compounds was significantly decreased at 0.18 mg/cm². In a fumigation test with S. oryzae adults, bornyl acetate and terpinolene were much more effective in sealed containers than in open ones, indicating that the insecticidal activity of these compounds was largely attributable to fumigant action. These naturally occurring materials derived from leaves of Chamaecyparis could be useful for managing populations of C. chinensis and S. oryzae.     

Changes in fatty acid and essential oil composition of sage (Salvia officinalis L.) leaves under NaCl stress

Fatty acids and essential oils from hydroponically cultivated Salvia officinalis leaves were analyzed by GC and GC–MS. Four different levels of NaCl (25, 50, 75 and 100 mM) were applied. The first results showed that salt treatment reduced significantly the plant growth by 61% and the total fatty acids (TFA) content by 32% at 100 mM NaCl. Alpha-linolenic, gadoleic, palmitic and oleic acids were the major fatty acids. Moreover, the polyunsaturated fatty acids decreased, while the monounsaturated ones increased with respect to increasing salinity. Regarding the essential oil composition, the main compounds were α-and β-thujone, 1,8-cineole, camphor, α-humulene, viridiflorol and manool at all salt treatments. The yield had a maximum increase at 75 mM NaCl. Hence, sage can be considered as moderately salt sensitive.     

Sacha inchi (Plukenetia volubilis): A seed source of polyunsaturated fatty acids,tocopherols, phytosterols,phenolic compounds and antioxidant capacity

Fatty acids (FA), phytosterols, tocopherols, phenolic compounds, total carotenoids and hydrophilic and lipophilic ORAC antioxidant capacities were evaluated in 16 cultivars of Sacha inchi (SI) seeds with the aim to valorise them and offer more information on the functional properties of SI seeds. A high α linolenic (α-Ln) fatty acid content was found in all cultivars (ω3, 12.8–16.0 g/100 g seed), followed by linoleic (L) fatty acid (ω6, 12.4–14.1 g/100 g seed). The ratio ω6/ω3 was within the 0.83–1.09 range. γ- and δ-tocopherols were the most important tocopherols, whereas the most representative phytosterols were β-sitosterol and stigmasterol. Contents of total phenolics, total carotenoids and hydrophilic and lipophilic antioxidant capacities ranged from 64.6 to 80 mg of gallic acid equivalent/100 g seed; from 0.07 to 0.09 mg of β-carotene equivalent/100 g of seed; from 4.3 to 7.3 and, from 1.0 to 2.8 μmol of Trolox equivalent/g of seed, respectively, among the evaluated SI cultivars. Results showed significant differences (p < 0.05) among the evaluated SI cultivars in the contents of ω3, ω6, antioxidant capacities and other evaluated phytochemicals. SI seeds should be considered as an important dietary source of health promoting phytochemicals.     

Supercritical CO2 cell breaking extraction of Lycium barbarum seed oil and determination of its chemical composition by HPLC/APCI/MS and antioxidant activity

The extraction parameters for oil extraction from Lycium barbarum seed including extraction pressure, temperature and time were optimized using an orthogonal test design. The optimum conditions for supercritical CO₂ extraction were as follows: extraction pressure, 30 MPa; extraction temperature, 45 °C; dynamic extraction time, 60 min; CO₂ flow, 25 kg/h. The oil yield under the conditions proposed was 19.28 g/100 g. The effect of cell wall breakage pretreatment was investigated by supercritical CO₂ rapid depressurization, and results indicated this pretreatment could result in a rapid and efficient extraction. A sensitive fluorescent reagent 2-(11H-benzo[a]carbazol-11-yl) ethyl 4-methylbenzenesulfonate (BCETS) was utilized as pre-column labeling regent to determine fatty acids (FA) from Lycium barbarum seed oils obtained by different extraction methods. The main FA were: C18:2, C18:1, C16, C20:6, C18:3, and C20. The oil from L. barbarum seed exhibited excellent antioxidant activity in 2,2-diphenyl-1-picrylhydrazyl（DPPH）radical scavenging assay and β-carotene bleaching test, and its antioxidant activity compared well with the references ascorbic acid and α- tocopherol.     

Isolation and chemical evaluation of carob (Ceratonia siliqua L.) seed germ

The present investigation was undertaken in order to study the influence of the dehusking procedure on the germ meal composition of carob (Ceratonia siliqua L.) seeds and also to investigate its detailed composition and nutritive value. Meals of carob seed germ were obtained by acid treatment or boiling water treatment of the whole seeds. These procedures allowed to separation of the tight-fitting brown coat of the seed and the removal of the endosperm. Results indicated that the carob germ meal composition could be affected by the isolation procedures. Small reductions were observed in protein and lipid contents in germ meals from acid extraction. The analysis of the carob germ meal (containing fine fragments of husk and endosperm), which could be really obtained industrially, showed the following composition: moisture 8.3%, ash 6.5%, lipids (neutral and polar) 6.6% containing ∼21% of polar lipids, crude proteins 54.7% and energy value 17.5 kJ/g.