Danilo Falconieri

Extraction and Separation of Volatile and Fixed Oils from Seeds of Myristica fragrans by Supercritical CO2: Chemical Composition and Cytotoxic Activity on Caco-2 Cancer Cells

UNLABELLED Isolation of volatile and fixed oils from nutmeg have been obtained by supercritical fractioned extraction with carbon dioxide. Extraction experiments were carried out at pressures of 90 and 250 bar and temperature of 40 °C. The extraction step performed at 90 bar produced a volatile fraction mainly formed by myristicin (32.8%), sabinene (16.1%), α-pinene (9.8%), β-pinene (9.4%), β-phellandrene (4.9%), safrole (4.1%) and terpinen-4-ol (3.6%). The oil yield relative to this step of the process was 1.4% by weight of the charge. The last extraction step at 250 bar produced a butter-like material (nutmeg butter). The yield of this step was 14.4% by weight. The most represented fatty acids of fixed oil from nutmeg were 14:0 (79.2%), 18:1 n-9 (7.4%) and 16:0 (6.1%), and in particular the unsaturated fatty acids 18:1 n-9 averaged 32.96 μg/mg of oil. The level of myristicin in the nutmeg essential and fixed oils was also directly quantified by reversed HPLC-DAD. Moreover, the essential oil obtained from nutmeg, as well as myristicin, showed a significant in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells). PRACTICAL APPLICATION In this study, the chemical characterization and the anticancer activity of nutmeg oils obtained by supercritical extraction with carbon dioxide were investigated. This is important for their potential application in food and pharmaceutical industries.

Extraction of Oil from Wheat Germ by Supercritical CO2

This study examined the supercritical fluid extraction of wheat germ oil. The effects of pressure (200-300 bar at 40 °C) and extraction time on the oil quality/quantity were studied. A comparison was also made between the relative qualities of material obtained by SFE and by organic solvent extraction. The extracts were analyzed for α-tocopherol and polyunsaturated fatty acid content. The maximum wheat germ oil yield at about 9% was obtained with supercritical carbon dioxide extraction at 300 bar, while fatty acid and α-tocopherol composition of the extracts was not remarkable affected by either pressure or the extraction method.

Effect of acute administration of Pistacia lentiscus L. essential oil on rat cerebral cortex following transient bilateral common carotid artery occlusion

BackgroundIschemia/reperfusion leads to inflammation and oxidative stress which damages membrane highly polyunsaturated fatty acids (HPUFAs) and eventually induces neuronal death. This study evaluates the effect of the administration of Pistacia lentiscus L. essential oil (E.O.), a mixture of terpenes and sesquiterpenes, on modifications of fatty acid profile and endocannabinoid (eCB) congener concentrations induced by transient bilateral common carotid artery occlusion (BCCAO) in the rat frontal cortex and plasma.MethodsAdult Wistar rats underwent BCCAO for 20 min followed by 30 min reperfusion (BCCAO/R). 6 hours before surgery, rats, randomly assigned to four groups, were gavaged either with E.O. (200 mg/0.45 ml of sunflower oil as vehicle) or with the vehicle alone.ResultsBCCAO/R triggered in frontal cortex a decrease of docosahexaenoic acid (DHA), the membrane highly polyunsaturated fatty acid most susceptible to oxidation. Pre-treatment with E.O. prevented this change and led further to decreased levels of the enzyme cyclooxygenase-2 (COX-2), as assessed by Western Blot. In plasma, only after BCCAO/R, E.O. administration increased both the ratio of DHA-to-its precursor, eicosapentaenoic acid (EPA), and levels of palmytoylethanolamide (PEA) and oleoylethanolamide (OEA).ConclusionsAcute treatment with E.O. before BCCAO/R elicits changes both in the frontal cortex, where the BCCAO/R-induced decrease of DHA is apparently prevented and COX-2 expression decreases, and in plasma, where PEA and OEA levels and DHA biosynthesis increase. It is suggested that the increase of PEA and OEA plasma levels may induce DHA biosynthesis via peroxisome proliferator-activated receptor (PPAR) alpha activation, protecting brain tissue from ischemia/reperfusion injury.

Antibacterial, allelopathic and antioxidant activities of essential oil of Salvia officinalis L. growing wild in the Atlas Mountains of Morocco.

Salvia officinalis (Common sage, Culinary sage) is an aromatic plant that is frequently used as a spice in Mediterranean cookery and in the food industry and as a traditional medicine for the treatment of several infectious diseases. The essential oils were obtained by two different methods [hydrodistillation (HD) and microwave (Mw)] from the aerial part of S. officinalis L. growing wild in Ourika – Marrakech in Morocco. Ourika is a large zone of the Atlas Mountains which is considered as a large reserve of Flora, especially medicinal and aromatic plants. The obtained oils were analysed by gas chromatography and gas chromatography–mass spectrometry and compared with that of Tunisia. Thirty-six compounds were identified from the Mw-extracted oil which accounted for 97.32% of the total oil composition. However, 33 compounds obtained by HD representing 98.67%. The major components were trans-thujone (14.10% and 29.84%), 1,8-cineole (5.10% and 16.82%), camphor (4.99% and 9.14%), viridiflorol (16.42% and 9.92%), β-caryophyllene (19.83% and 5.20%) and α-humulene (13.54% and 4.02%). Antibacterial, allelopathic (% germination in lettuce seeds and inhibited root growth obtained after treatment with S. officinalis oils) and antioxidant (IC50 values 22 mg/mL) activities were studied.

Chemical composition and biological assays of essential oils of Calamintha nepeta (L.) Savi subsp. nepeta (Lamiaceae)

Aerial parts of wild Calamintha nepeta (L.) Savi subsp. nepeta growing spontaneously on the Mediterranean coast (Sardinia Island, Italy) and on the Atlantic coast (Portugal) were used as a matrix for the supercritical extraction of volatile oil with CO2. The collected extracts were analysed by GC-FID and GC-MS methods and their compositions were compared with that of the essential oil isolated by hydrodistillation, but the differences were not relevant. A strong chemical variability was observed in the essential oils depending on the origin of the samples. The results showed the presence of two chemotypes of C. nepeta. In all Italian samples, pulegone, piperitenone oxide and piperitenone were the main components (64.4–39.9%; 2.5–19.1%; 6.4–7.7%); conversely, the oil extracted from Portuguese C. nepeta is predominantly composed of isomenthone (35.8–51.3%), 1,8-cineole (21.1–21.4%) and trans-isopulegone (7.8–6.0%). The minimal inhibitory concentration (MIC) and the minimal lethal concentration (MLC) were used to evaluate the antifungal activity of the oils against Candida albicans, Candida tropicalis, Candida krusei, Candida guillermondii, Candida parapsilosis, Cryptococcus neoformans, Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Microsporum gypseum, Epidermophyton floccosum, Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus. The Italian oil, rich in pulegone, exhibited significant antifungal activity against Aspergillus and dermatophyte strains, with MIC values of 0.32–1.25 µL mL−1.

Isolation of the volatile fraction from Apium graveolens L. (Apiaceae) by supercritical carbon dioxide extraction and hydrodistillation: Chemical composition and antifungal activity

Apium graveolens L. (wild celery), belonging to the family of Apiaceae, is a scaposus hemicryptophyte. Instead, the cultivate plant is an annual or biennial herb widely used as a spice and seasoning in food. A broad range of biological activities have been attributed to A. graveolens. These include antimicrobial activity, larvicidal activity, hepatoprotective activity, nematicidal and mosquito repellent potential and antihyperlipidaemic properties.In this study, the authors compare the composition of the volatile fractions of A. graveolens collected in natural populations in Portugal and Italy and evaluate their potential as antifungal agents.The composition of the volatile oils obtained by hydrodistillation and their antifungal activity are reported. The oils were analysed by gas chromatography–flame ionisation detector and gas chromatography–mass spectrometry methods and their composition were compared with that of the volatile extracts isolated by supercritical CO2. A chemical variability in the extracts depending on the origin of the plants and on the extraction method was observed. The results showed the presence of sedanenolide, neocnidilide and neophytadiene as main components. The minimal inhibitory concentration (MIC) and the minimal lethal concentration were used to evaluate the antifungal activity of the oils against Candida albicans, Candida tropicalis, Candida krusei, Candida guilliermondii, Candida parapsilosis, Cryptococcus neoformans, Trichophyton rubrum, Trichophyton mentagrophytes, T. mentagrophytes var. interdigitale, Trichophyton verrucosum, Microsporum canis, Microsporum gypseum, Epidermophyton floccosum, Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus. The oil from Italy rich in neophytadiene is the more active, with MIC values of 0.04–0.64 µL mL−1. Our results show that A. graveolens volatile extracts may be useful in the clinical treatment of fungal diseases.

Faceted phospholipid vesicles tailored for the delivery of Santolina insularis essential oil to the skin

The aim of this work was to formulate Santolina insularis essential oil-loaded nanocarriers, namely Penetration Enhancer containing Vesicles (PEVs), evaluate the physico-chemical features and stability, and gain insights into their ability to deliver the oil to the skin. S. insularis essential oil was obtained by steam distillation, and was predominantly composed of terpenes, the most abundant being β-phellandrene (22.6%), myrcene (11.4%) and curcumenes (12.1%). Vesicles were prepared using phosphatidylcholine, and ethylene or propylene glycol were added to the water phase (10% (v/v)) to improve vesicle performances as delivery systems. Vesicles were deeply characterized by light scattering, cryogenic transmission electron microscopy and small/wide-angle X-ray scattering, the results showing polyhedral, faceted, unilamellar vesicles of ∼115 nm in size. The presence of the glycols improved vesicle stability under accelerated ageing conditions, without changes in size or migration phenomena (e.g. sedimentation and creaming). Confocal laser scanning microscopy images of pig skin treated with S. insularis formulations displayed a penetration ability of PEVs greater than that of control liposomes. Moreover, all formulations showed a marked in vitro biocompatibility in human keratinocytes. These findings suggest that the nanoformulation may be of value in enhancing the delivery of S. insularis essential oil to the skin, where it can exert its biological activities.

Essential oil composition of leaves of Stachys yemenensis obtained by supercritical CO2

This article reports the composition of the essential oil from the leaves of Stachys yemenensis. The essential oil was extracted by supercritical CO2 (90 bar; 40°C) and its chemical composition was determined by gas chromatography and gas chromatography–mass spectrometry. The major components of the sample were α-phellandrene (13.9%), β-phellandrene (11.7%), elemol (12.0%), spathulenol (6.7%), β-eudesmol (5.0%), α-eudesmol (4.75%) and squalene (4.8%). On the exhausted matrix, deprived of the volatiles, we carried out a high-pressure (250 bar) treatment for the extraction of squalene (49.7%). The antimicrobial activity of the essential oils has been assayed by using the broth dilution method on two American Type Culture Collection (ATCC) strains, Escherichia coli ATCC 35218 and Staphylococcus aureus ATCC 43300, and two clinical strains, Candida albicans and Candida glabrata.

Extraction, separation and isolation of volatiles from Vitex agnus-castus L. (Verbenaceae) wild species of Sardinia, Italy, by supercritical CO2

Isolation of volatile concentrates from leaves, flowers and fruits of Vitex agnus-castus L. have been obtained by supercritical extraction with carbon dioxide. The composition of the volatile concentrates has been analysed by GC/MS. In all plant organs, the extracts are composed chiefly of α-pinene, sabinene, 1,8-cineole, α-terpinyl acetate, (E)-caryophyllene, (E)-β-farnesene, bicyclogermacrene, spathulenol and manool. The main difference observed was in the content of sclarene, which was not present in the samples from flowers or fruits. To complete the investigation, a comparison with the hydrodistilled oil has been carried out. The minimal inhibitory concentration (MIC) and the minimal lethal concentration were used to evaluate the antifungal activity of the oils against dermatophyte strains (Trichophyton mentagrophytes, Microsporum canis, T. rubrum, M. gypseum and Epidermophyton floccosum). Antifungal activity of the leaf essential oil was the highest, with MIC values of 0.64 µL mL−1 for most of the strains.

Chemical composition and antifungal activity of supercritical extract and essential oil of Tanacetum vulgare growing wild in Lithuania

This article reports the results concerning the qualitative and quantitative composition of the volatile oil obtained from the aerial parts of Tanacetum vulgare, collected in Lithuania, by supercritical fluid extraction (SFE) with CO2 at 90 bar and 40 °C and by hydrodistillation (HD). GC-MS and GC-FID analysis revealed that the oils mainly consisted of 1,8-cineole, trans-thujone, cis-chrysanthenol, borneol and myrtenol. Oils obtained by HD displayed, with respect to the SFE volatile oil, a higher amount of hydrocarbon monoterpenes. The hydrodistilled oil was particularly active against dermatophyte strains and Cryptococcus neoformans, with MIC values in the range of 0.16–0.64 μL/mL. The oil revealed a significant inhibitory effect on germ tube formation in Candida albicans at sub-inhibitory concentrations. At the concentration of 1/16 MIC, the inhibition of filamentation was more than 80% in comparison to untreated control cells.