Composition and biological activity of the essential oil from Thapsia minor, a new source of geranyl acetate

The essential oils from aerial parts of Thapsia minor Hoffmanns. & Link (Apiaceae), obtained by hydrodistillation, were analyzed by gas-chromatography and gas-chromatography-mass spectrometry. Oxygenated monoterpenes represented the main fraction with geranyl acetate (82.3-83.0%) as the major constituent, making this plant a good source of this compound.Biological activity of the essential oil was evaluated against several fungal strains. The highest activity was found against dermatophyte strains, Crytococcus neoformans and Candida guillermondii with minimal inhibitory concentrations values from 0.32 μl/ml to 1.25 μl/ml. The antifungal activity could be associated with the significant contribution of the oxygenated monoterpene geranyl acetate.Furthermore, this essential oil demonstrated to have potent anti-inflammatory in vitro activity, at concentrations that did not affect macrophages and keratinocytes viability, being the anti-inflammatory properties of the oil mainly due to geranyl acetate.These favorable results emphasize the antifungal and anti-inflammatory abilities of the molecule geranyl acetate and further support the exploitation of T. minor oil for pharmaceutical industry, in order to find new phytopharmaceuticals.     

Effect of residual distillation water of 15 plants and three plant hormones on Scotch spearmint (Mentha × gracilis Sole)

Distillation waste water is a byproduct from steam distillation of aromatic crops, and is currently discharged into streams and rivers. We evaluated distillation waste water (extract) from 15 essential oil crops plus three plant hormones (methyl jasmonate, MJ; gibberellic acid, GA3; and salicylic acid, SA) as foliar spray for Scotch spearmint (Mentha × gracilis Sole). GA3 and Achillea millefolium extract decreased essential oil content. Hypericum perforatum extract increased α-pinene, whereas SA decreased it. H. perforatum extract increased β-pinene and sabinene concentrations relative to hormones but was not different from the control. H. perforatum also increased l-limonene, while SA and GA3 decreased myrcene and MJ and SA decreased l-limonene. Application of MJ and SA increased l-carvone concentration relative to the control and most other treatments. The sulfur concentration in plant extracts was positively correlated to β-caryophyllene. None of the residual distillation waters showed significant antimicrobial or antimalarial activity. The distillation waste water from essential oil crops may serve as a modifier for Scotch spearmint essential oil.     

Preparation and evaluation of antioxidant capacity of Jackfruit (Artocarpus heterophyllus Lam.) wine and its protective role against radiation induced DNA damage

Jackfruit is an underutilized edible fruit in the tropics and subtropics. The aim of this study was to produce wine from jackfruit pulp and to evaluate the total phenolic and flavonoid contents and antioxidant properties of the wine. The ability of scavenging free radicals was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant assay (FRAP), N, N-dimethyl-p-phenylendiamine (DMPD) and nitric oxide (NO) scavenging assays. Experimental results indicated that jackfruit wine was effective in DPPH radical scavenging (69.44 ± 0.34%), FRAP (0.358 optical density value, O.D.), DMPD (78.45 ± 0.05%) and NO (62.46 ± 0.45%) capacity. By the analysis of the high performance liquid chromatography coupled to diode array detector (HPLC-DAD), two phenolic compounds namely gallic acid and protocatechuic acid were identified. The jackfruit wine was also able to protect H₂O₂ + UV radiation and γ-radiation (100 Gy) induced DNA damage in pBR322 plasmid DNA. The antioxidant and DNA damage protecting properties of jackfruit wine confirmed health benefits when consumed and could become a valuable source of antioxidant rich neutraceuticals. Additionally, the wine could be a commercially valuable by-product for the jackfruit growers.     

Lavender and hyssop productivity, oil content, and bioactivity as a function of harvest time and drying

Field and laboratory experiments were conducted to evaluate the productivity and essential oil composition of lavender (Lavandula angustifolia Mill.) and hyssop (Hyssopus officinalis L.) as functions of year, harvest time, and drying. Lavender essential oil content ranged from 0.71 to 1.3% (overall average of 0.89%) and hyssop oil content ranged from 0.13 to 0.26% (overall average of 0.19%). Lavender and hyssop essential oil yields increased with time. Hyssop oil yields varied from 7.3 kg ha⁻¹ to 19.6 kg ha⁻¹, and lavender oil yields varied from 7.8 kg ha⁻¹ to 55.5 kg ha⁻¹. The major constituents of lavender oil were linalool (23.3-43.4%) and linalylacetate (20.2-39.6%), while the major constituents of hyssop oil were pinocamphene + isopinocamphene (57-75%) and β-pinene (5-15%). Lavender oil extracted from dry material had higher concentrations of linalyl acetate and caryophyllene but lower concentrations of myrcene than the oil from the fresh material. Delayed harvest of hyssop increased the concentrations of β-pinene, myrcene, and limonene + cineole but reduced pinocamphone + isopinocamphone. The chemical composition of the lavender and hyssop oil produced in Mississippi was similar to commercial oils from Bulgaria, Canada, France, and US. Lavender and hyssop can be established as essential oil crops in areas of the southeastern United States. Lavender and hyssop essential oils did not show significant antimicrobial, antileishmanial, antimalarial activity, and did not alter ruminal fermentation. However, commercial oil from L. latifolia reduced methane production in an in vitro digestibility study. The antioxidant activity of hyssop essential oil was 2039 μmol of TE L⁻¹, whereas the antioxidant activity of lavender essential oil was 328 μmol of TE L⁻¹.     

Enzyme inhibitory and antioxidant activity of Melia azedarach L. naturalized in Anatolia and its phenolic acid and fatty acid composition

In the present study, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase inhibitory and antioxidant activities of the fruit and leaf extracts of Melia azedarach L. (Meliaceae) of Turkish origin were evaluated. Enzyme inhibitory activity of the extracts was tested in vitro using ELISA microplate reader. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-chelation, and ferric-reducing antioxidant power (FRAP) assays. Phenolic composition of the extracts was elucidated by high performance liquid chromatography (HPLC) and fatty acid compositions of the fatty oils of the fruits and leaves were elucidated by gas chromatography-mass spectrometry (GC-MS). The ethyl acetate extract from the leaves showed the highest inhibition against AChE (33.63 ± 1.40%) and BChE (92.89 ± 3.05%). The methanol extract from the leaves exerted the best antioxidant activity in DPPH radical scavenging and FRAP assays, while the ethyl acetate extracts of the fruits and leaves had the most notable effect in metal-chelation assay.     

Jatropha curcas seed cake as substrate for production of xylanase and cellulase by Aspergillus niger FGSCA733 in solid-state fermentation

Jatropha curcas seed-cake was evaluated for use as a solid state fermentation substrate for production of cellulolytic and xylanolytic enzymes by Aspergillus niger. Supplementation of the seedcake with 10% thatch grass (Hyperrhaenia sp.) resulted in a fivefold increase in xylanase production. Ammonium chloride supplementation increased production of xylanase by 13%. Under the same conditions, cellulase production was not influenced by supplementation with grass or the nitrogen sources used. Maximum xylanase was produced at 25 °C whilst cellulase was maximally produced at 40 °C. Highest xylanase activity was obtained when the cultures had an initial pH of 3 whereas cellulase was maximally produced at an initial pH of 5. Under optimised conditions, 6087 U and 3974 U of xylanase and cellulase respectively were obtained per gram of substrate. Zymograms of crude enzyme extracts showed six active bands ranging from 20 kDa to 43 kDa for cellulase and a 31 kDa active band for xylanase.     

Comparative seasonal variation and chemical composition of essential oils from the leaves and stems of Schefflera heptaphylla using microwave-assisted and conventional hydrodistillation

Two different isolation techniques, microwave-assisted hydrodistillation (MAHD) and conventional hydrodistillation (HD), were used to extract the volatile constituents from the leaves and stems of Schefflera heptaphylla (L.) Frodin harvested during four different seasons, which were subsequently analyzed via gas chromatography with flame ionization detection and gas chromatography with mass spectrometry. MAHD is more advantageous than HD in terms of energy savings and extraction time (60 min vs. 180 min for MAHD vs. HD). Large variations in the composition among the different oils were observed. The seasonal changes in the percentages of the main constituents of the oils were irregular; this affected the quantity and composition of the oil. Oil yield was affected by the method of extraction and seasonal changes. It ranged from 0.11% to 0.27%, with the maximum amount of oil extracted from the leaves using MAHD in winter and the minimum from the stem oil extraction using HD in spring. The essential oil content varied significantly with the season. Sesquiterpene hydrocarbons constituted the major portion of the essential oils. Sesquiterpene hydrocarbons were highest in the leaf oil extraction by HD during winter (88.6%) and lowest in the stem oil extraction by HD during spring (56.3%). Microwave irradiation did not adversely affect the composition of the essential oils. MAHD, indeed, is a modern, green, and fast technology.     

Regulation of tissue differentiation by plant growth regulators on tTCLs of Panax ginseng adventitious roots

Differentiated tissue in Panax ginseng cultures was found to be very efficacious for saponin production. In order to increase the yield of saponins and preserve culture stability we were testing different plant growth regulators (PGR) and auxin/cytokinin combinations to regulate a level of tissue differentiation. For this purpose we used transverse thin cell layers (tTCLs) of adventitious roots of Panax ginseng. Adventitious roots were cultivated in Shenk and Hildebrand (SH) liquid medium supplemented with IBA (24.6 μM). Callus formation and root multiplication of adventitious root tTCLs was evaluated after 4 and following 12 weeks of cultivation, respectively, on SH basal medium containing various auxins (3 mg l⁻¹) or cytokinins (0.2 or 0.02 mg l⁻¹) or their combinations. We found that kinetin (Kin) in combination with auxin benzo[b]selenienyl acetic acid (BSAA), naphthalene acetic acid or indole-3-butric acidis the best for biomass production and following root multiplication. These combinations were tested in previously selected most suitable large-scale system—a temporary immersion system RITA. The best saponin production (15.94 ± 1.89 mg g⁻¹ dry weight) and growth value (5.62 ± 0.34) was reached on medium containing BSAA and Kin combination.     

Carbohydrate degrading enzyme production by plant pathogenic mycelia and microsclerotia isolates of Macrophomina phaseolina through koji fermentation

The ability of hydrolytic enzyme production by two different isolates of Macrophomina phaseolina was studied on apple pomace as a substrate for solid state fermentation (SSF). Initial moisture level, temperature and fermentation period was optimized so as to achieve higher output. Among the two different isolates, microsclerotial (MphP) and mycelial (MphM), MphP was observed as a potential source of different hydrolytic enzymes as compared to MphM. MphP gave higher enzyme activities (IU/gram dry substrate (gds): filter paper cellulase (FPase) activity 196.21 ± 16.3 (120 h), carboxymethyl cellulase (CMCase) 279.34 ± 28.25 (72 h), β-glucosidase (BGL) 129.82 ± 12.41 (96 h), xylanase 2527.88 ± 46.15 (120 h), and amylase 2780.72 ± 38.13 (96 h), respectively at 70% (v/w) IML. The incubation temperature was also found to have impact on the enzyme production ability of Macrophomina strains. The higher enzyme activities were achieved (IU/gds) as follows FPase 276.13 ± 25.02 (40 °C, 120 h), CMCase 278.11 ± 24.47 (35 °C, 144 h), BGL 189.47 ± 15.05 (30 °C, 144 h), xylanase 3845.77 ± 43.38 (35 °C, 144 h) and amylase activity of 3309.45 ± 29.22 (35 °C, 120 h), respectively using MphP at 70% (v/w) IML. This study reports for the first time the potential of carbohydrate degrading enzyme bioproduction by different isolates of M. phaseolina.     

Effect of extraction methods on characteristic and composition of Indonesian cashew nut shell liquid

Long chain phenols contained in cashew nut shell liquid (CNSL) are found to have important pharmaceutical applications, such as antitumor, antimicrobial, urease inhibitory and lipoxygenase activities, and also are well known in coating and resin industry. The impact of different extraction methods on CNSL yield, selectivity towards preferable compounds, composition and characteristic of extracts was investigated. Four different methods employed in extracting CNSL from CNS were: supercritical carbon dioxide (SC-CO₂) extraction (40 °C, 300 bar, 4 h), subcritical water (SCW) extraction (140 °C, 22 bar, 1 h), soxhlet extraction (solvent boiling point, atmospheric pressure, 30 h), and two-step extraction, which comprises a solvent extraction followed by a SCW extraction. Characteristic of the extracts differed significantly. Methanol and two-step extraction resulted in darker and more turbid extracts, while n-hexane and SC-CO₂ extracts were clearer and lighter in color. GC-FID/MS chromatograms showed differences in compositions of the extracts obtained by different methods. Two-step extraction yielded extracts that contain 81.17-82.98% total long chain phenols (around 50% based on dry CNS) with monounsaturated cardanol as the major compound, producing higher amount of total phenols than other methods. SCW extraction showed high selectivity towards monounsaturated cardanol and stigmasterol, while high concentration of monounsaturated anacardic acid and cardol appeared in SC-CO₂ extract.     

Enzymatic epoxidation of Sapindus mukorossi seed oil by perstearic acid optimized using response surface methodology

As a novel renewable resource, Sapindus mukorossi seed oil (SMSO) with an iodine value of 84.86 g/100 g, and containing 51.0 ± 0.9% oleic acid (18:1), 6.6 ± 0.6% linoleic acid (18:2), 1.1 ± 0.3% linolenic acid (18:3), and 23.1 ± 0.9% eicosanoic acid (20:1), was epoxidized using hydrogen peroxide as oxygen donor and stearic acid as active oxygen carrier in the presence of immobilized Candida antarctica lipase B. The effect of the amount of stearic acid on the enzymatic epoxidation was investigated. Response surface methodology (RSM) was used to study and optimize the effects of variables (reaction temperature, enzyme load, mole ratio of H₂O₂/CC-bonds, and reaction time) on the epoxy oxygen group content (EOC) of epoxidized SMSO. Results showed that stearic acid as active oxygen carrier could enhance the enzymatic epoxidation of SMSO. The variables of reaction temperature and enzyme load were the most significant in the process. A two second-order model was satisfactorily fitted the data (R² = 0.9723) with non-significant lack of fit. The optimum EOC of epoxidized SMSO was 4.6 ± 0.3% under the conditions of 50.0 °C, 7.0 h, 2.00% (relative to the weight of SMSO) enzyme load, and 4:1 mole ratio of H₂O₂/CC-bonds.     

Genotype by environment interaction of seed and oil yield in vernonia (Vernonia galamensis variety ethiopica)

Vernonia (Vernonia galamensis variety ethiopica) is a potentially novel renewable source of natural epoxy oil. The objective of this study was to determine the genotype by environment interaction and to identify superior and stable genotypes of vernonia with high seed and oil yield. Field experiments were conducted during 2006, 2007 and 2008 at two localities namely, Gabaza and Syferkuil. Ten selected genotypes of vernonia were evaluated for seed yield, oil content and oil yield using the randomized complete block design with three replications. Significant interactions (P ≤ 0.05) were detected among genotype by location for seed yield, oil content, oil yield and genotype by year for seed and oil yield. Genotype Vge-18 had the highest seed yield ranging between 3095 and 3337 kg/ha followed by Vge-17 yielding 3006-3137 kg/ha at Gabaza. These genotypes were also the best performers at Syferkuil where Vge-17 yielded 2915-3217 kg/ha followed by Vge-18 with 2819-3152 kg/ha. The superiority statistics allocated Vge-17 and Vge-18 as best yielding and stable genotypes. In both locations Vge-4 had increased seed oil contents at 43% (Gabaza) and 35% (Syferkuil). Other promising genotypes with high seed oil content were Vge-33 at Gabaza (40-43%) and Vge-3 at Syferkuil (34-35%). Genotypes with the highest oil yields were found to be Vge-18 (1117-1370 kg/ha) at Gabaza and Vge-4 with yields of 885-922 kg/ha at Syferkuil. Overall, Vge-17 and Vge-18 were identified as having the highest seed yield, while Vge-4 and Vge-3 yielded the highest seed oil content with average stability. These genotypes could be used for direct large scale production or strategic breeding of vernonia in these or similar environments.     

Microscopy techniques and image analysis for evaluation of some chemical and physical properties and morphological features for seeds of the castor oil plant (Ricinus communis)

Ricinus communis seed is a source of protein and oil with a high potential to use as animal's feedstock and biodiesel production. However, the oil yield and the extraction efficiency depend on the process conditions applied, as well as on the physical, chemical and structural properties of the seed, which have not been fully investigated. Hence, the objective of this study was to evaluate some chemical and physical properties of R. communis seed as well as to describe and quantify the macro and microstructure of this raw material by microscopy techniques and image analysis. Chemical analysis confirmed the seeds’ high contents of protein (28.48 ± 0.25%) and fat (51 ± 0.31%). On the other hand, the values of geometric mean diameter (8.95 ± 0.05 mm), bulk density (538 ± 11 kg/m³), and true density (1458 ± 27 kg/m³), among others, were higher than the ones reported about similar oils seeds. Microstructural studies showed that the endosperm cells presented an ovoid shape, as obtained from the aspect ratio results (AR = 1.28 ± 0.17), and a cell density of 570 ± 10 cell/mm², resulting in a porous structure, while the embryo cells had a cell density of 4903 ± 2 cell/mm², and an AR of 2.41 ± 0.48, related to a more compact structure (rectangular form) in this part of the seed. Regarding to lipids bodies (lb), they were only visible in the endosperm cells, showing a circular shape (AR = 1.16 ± 0.1), and a mean cell density of 9.57 ± 2.40 lipid bodies/μm², associated to protein as observed by the mineral presence (K, P, Mg and S) as determined by the energy dispersive X-ray analysis. Microscopy techniques and images analysis were efficient tools for the characterization of macro and microstructure of seeds and the data obtained integrate numerical information that could be useful for thermal and mechanical processing of R. communis seed, as well as for the design process equipment.     

Screening and quantification of an antiseptic alkylamide, spilanthol from in vitro cell and tissue cultures of Spilanthes acmella Murr.

The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this, in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots were induced in the presence of N⁶-benzylaminopurine (BAP) alone or in combination with either α-naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) in Murashige and Skoog medium. The best treatment for shoot regeneration was MS + BAP (5.0 μM) + IAA (5.0 μM), which promoted adventitious shoot proliferation in >82% cultures with an average of 5.3 shoots per explant. Regenerated shoots rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to half strength) containing 50 g l⁻¹ sucrose. The plantlets were acclimatized successfully with 90% survival rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation pattern. For quantification studies, calibration curve was generated, which revealed a higher amount of spilanthol content (3294.36 ± 12.4 μg/g DW) in the leaves of in vitro plants compare to those of in vivo plants (2703.66 ± 9.6 μg/g DW of spilanthol). An efficient multiplication frequency, ploidy stability and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially important medicinal plant.     

Comparative assessment of antioxidant and cholinesterase inhibitory properties of the marigold extracts from Calendula arvensis L. and Calendula officinalis L.

Calendula sp. is an important medicinal and industrial plant with various bioactivities. In this study, we examined enzyme inhibitory effects of the n-hexane, dichloromethane, acetone, ethyl acetate, methanol, and water extracts of the leaf and flowers of Calendula arvensis L. and C. officinalis L. against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) assays at 250, 500, and 1000 μg mL⁻¹. Total phenol and flavonoid quantification of the extracts was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The ethyl acetate extract of C. arvensis flowers was the most active in AChE inhibition assay (31.24 ± 1.29%), while the n-hexane extract of C. officinalis leaves exerted the highest ferric ion-chelating capacity (74.27 ± 2.25%). Thin layer chromatographic analysis indicated presence of flavonoid and triterpene derivatives mainly in the extracts.     

Nitrogen applications modify seed and oil yields and fatty acid composition of winter mustard

Winter mustard (Brassica juncea L.) is not a common crop in the Southeastern United States. With increased interest in biodiesel production, there has been corresponding interest in mustard in this region. The objective of this study was to evaluate the effect of N fertilization (0, 50, 100, 150 kg N ha⁻¹) on productivity, oil content, and oil composition of winter mustard ‘Pacific Gold’ grown at three locations in Mississippi (Stoneville, and two locations at Verona, namely Verona silt loam (Verona-SL) and Verona clay (Verona-C)). Nitrogen did not affect oil content (percent oil). Seed and oil yields (kg ha⁻¹) increased with N application relative to the unfertilized control. At the Verona-C location, the concentration of oleic acid was higher in the 50 kg N ha⁻¹ treatment. At Stoneville, linolenic acid concentration was higher in the 150 kg N ha⁻¹ and lower in the 100 kg/N ha⁻¹ treatment, while it was not different in the other treatments. Overall, the yield of the fatty acids (FA) palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, arachidic, eicosanoic, behenic, erucic, lignoceric, and nervonic acid increased with higher N rates (100 or 150 kg N/h). The highest yield of FA in the two Verona locations were achieved in the 100 kg N ha⁻¹, while greatest yield of FA at Stoneville was achieved in the highest N rate (150 kg N ha⁻¹). Means of mustard oil yields in our study in the higher fertility treatment ranged from 737 to 1094 kg ha⁻¹. This study demonstrated winter mustard production in Mississippi and possibly other areas in the Southeastern United States can be successful and could provide seed and oil yields comparable to yields from other production areas.     

Bioactive properties and chemical constituents of methanolic extract and its fractions from Jatropha curcas oil

The present work is designed to evaluate the bioactive properties of the crude methanolic extract of Jatropha curcas oil and its solvent fractions. The crude methanolic extract obtained was fractionated using a hydrophilic lipophilic balanced (HLB) cartridge and then eluted with different solvents in the order of hexane (F1), dichloromethane (F2), chloroform (F3), ethyl acetate (F4) and methanol (F5), respectively. Total phenolic content of the crude methanolic extract and its fractions was in the range of 0.19-4.5 mg/g as gallic acid equivalent. Antioxidant activity of the crude methanolic extract and its fractions were determined by two complementary test methods, namely, phenanthroline method and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. All samples demonstrated weak antioxidant activity (150-851 μmol Fe/100 g of the extract and IC₅₀ of 1.05-13.5 mg/mL). When compared to butylated hydroxytoluene (BHT), a reference synthetic antioxidant, both showed weaker antioxidative potential. The evaluation of antimicrobial activity of the extracts was performed using a disc diffusion method and a micro-well dilution method against six economic plant disease bacteria. The results showed that all extracts possessed strong to moderate antibacterial activity with varying degrees of growth inhibition against the test bacteria. The minimum inhibitory concentrations (MIC) were in the range of 14.92-428.6 μg/mL. In addition, the chemical constituents in each fraction of the extract were subjected to analyze by gas chromatography-mass spectrometry (GC-MS). The eleven constituents were identified. Among them, 2,4-di-tert-butylphenol, methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate and linoleic acid may be the main cause of its strong antibacterial activity. Therefore, this oil present in the methanolic extract had great potential as effective antibacterial sources.     

Fatty acid composition and oxidation stability of hemp (Cannabis sativa L.) seed oil extracted by supercritical carbon dioxide

Supercritical carbon dioxide (SC-CO₂) was employed to extract oil from hemp (Cannabis sativa L.) seeds. For ground seeds, the supercritical extraction was carried out at temperatures of 40, 60 and 80 °C and pressures of 300 and 400 bar. Different solvent-ratios were applied. Supercritical CO₂ extractions were compared with a conventional technique, n-hexane in Soxhlet. The extraction yields, fatty acid composition of the oil and oxidation stability were determined. The seed samples used in this work contained 81% PUFAs, of which 59.6% was linoleic acid (ω-6), 3.4% γ-linolenic (ω-3), and 18% α-linolenic (ω-6). The highest oil yield from seeds was 22%, corresponding to 72% recovery, at 300 bar and 40 °C and at 400 bar and 80 °C. The highest oxidation stability corresponding to 2.16 mM Eq Vit E was obtained at 300 bar and 80 °C.     

Antifungal and antiaflatoxigenic efficacy of Caesulia axillaris Roxb. essential oil against fungi deteriorating some herbal raw materials, and its antioxidant activity

The study deals with evaluation of antifungal and antiaflatoxigenic Caesulia axillaris Roxb. essential oil (EO) against herbal raw materials deteriorating fungi and its free radical scavenging activity. During mycoflora analysis these herbal raw materials were found to be severely contaminated by different fungi and aflatoxins. A total of nine different fungal species were isolated from three herbal raw materials. Aspergillus flavus LHPtc was recorded as the highest aflatoxin B1 producing strain. EOs of some plants were tested for their fungitoxicity against the toxigenic strain A. flavus LHPtc, and C. axillaris EO was found as potent fungitoxicant. C. axillaris EO was chemically characterized through GC-MS analysis which depicted the presence of 18 compounds, dl-limonene and Euasarone being the major components. The EO exhibited broad spectrum of fungitoxicity against fungi causing postharvest deterioration of herbal raw materials. At 1.0 μl ml⁻¹ the oil showed complete inhibition of fungal growth and aflatoxin B₁ production was inhibited at 0.8 μl ml⁻¹. Free radical scavenging activity of the oil was also recorded by 2,2-diphenyl-1-picrylhydrazyl assay, and its IC₅₀ value was found 18 μl ml⁻¹. The safety limit of the EO was determined in terms of LD₅₀ on mice, which was 9166.6 μl kg⁻¹, suggesting its non mammalian toxicity. The EO of C. axillaris may be recommended as a plant based preservative in enhancement of shelf life of herbal raw materials by preventing their lipid peroxidation as well as biodeterioration due to fungal and aflatoxin contamination.     

Biological Properties of Fucoxanthin in Oil Recovered from Two Brown Seaweeds Using Supercritical CO2 Extraction

The bioactive materials in brown seaweeds hold great interest for developing new drugs and healthy foods. The oil content in brown seaweeds (Saccharina japonica and Sargassum horneri) was extracted by using environmentally friendly supercritical CO₂ (SC-CO₂) with ethanol as a co-solvent in a semi-batch flow extraction process and compared the results with a conventional extraction process using hexane, ethanol, and acetone mixed with methanol (1:1, v/v). The SC-CO₂ method was used at a temperature of 45 °C and pressure of 250 bar. The flow rate of CO₂ (27 g/min) was constant for the entire extraction period of 2 h. The obtained oil from the brown seaweeds was analyzed to determine their valuable compounds such as fatty acids, phenolic compounds, fucoxanthin and biological properties including antioxidant, antimicrobial, and antihypertension effects. The amounts of fucoxanthin extracted from the SC-CO₂ oils of S. japonica and S. horneri were 0.41 ± 0.05 and 0.77 ± 0.07 mg/g, respectively. High antihypertensive activity was detected when using mixed acetone and methanol, whereas the phenolic content and antioxidant property were higher in the oil extracted by SC-CO₂. The acetone–methanol mix extracts exhibited better antimicrobial activities than those obtained by other means. Thus, the SC-CO₂ extraction process appears to be a good method for obtaining valuable compounds from both brown seaweeds, and showed stronger biological activity than that obtained by the conventional extraction process.