Essential Oils from Piper lhotzkyanum Kunth Leaves from Brazilian Atlantic Forest: Chemical Composition and Stability in Different Storage Conditions.

This work aimed to evaluate the impact of different storage conditions and light and temperature exposures on the visual aspect and chemical composition of the essential oil (EO) of Piper lhotzkyanum Kunth, obtained from leaves by hydrodistillation from a region of high altitude. For this purpose, aliquots of the EO were stored for up to 90 days (a) under a refrigerator condition of 5 ± 3°C, (b) under a long-term (LT) condition of 30 ± 2°C and 75 ± 5% relative humidity (RH) and an accelerated condition (AS) of 40 ± 2°C and 75 ± 5% RH, and (c) in a photostability test achieved in amber and colorless glass vials. The changes were monitored on days 0 (control), 60, and 90 for the refrigerator, LT, and AS conditions. All EO chemical analyses were assessed by GC-FID and GC-MS for quantification and identification, respectively. It is reported, for the first time, that the EO of P. lhotzkyanum is rich in the sesquiterpenes ß-elemene and α-zingiberene. No significant changes in the EO was observed, revealing a minimal impact of temperature on the sample at the different storage conditions. However, there was a change in the content of α-zingiberene to bicyclogermacrene after exposure to light. The visual appearance of the samples was altered for all test conditions except the refrigerator condition. These results can potentially contribute to the product development of a bioactive EO from leaves of P. lhotzkyanum, a sesquiterpene rich natural material.

Effect of Enzymatic, Ultrasound, and Reflux Extraction Pretreatments on the Chemical Composition of Essential Oils.

The effect of different hydrodistillation pretreatments, namely, reflux extraction, reflux extraction with the addition of cell wall-degrading enzymes, and ultrasound, on the yield and chemical composition of essential oils of sage, bay laurel, and rosemary was examined. All pretreatments improved essential oil yield compared to no-pretreatment control (40-64% yield increase), while the oil quality remained mostly unchanged (as shown by statistical analysis of GC-MS results). However, enzyme-assisted reflux extraction pretreatment did not significantly outperform reflux extraction (no-enzyme control), suggesting that the observed yield increase was mostly a consequence of reflux extraction and enzymatic activity had only a minute effect. Thus, we show that ultrasound and reflux extraction pretreatments are beneficial in the production of essential oils of selected Mediterranean plants, but the application of enzymes has to be carefully re-evaluated.

Effects of light irradiation on essential oil biosynthesis in the medicinal plant Asarum heterotropoides Fr. Schmidt var. mandshuricum (Maxim) Kitag.

Asarum heterotropoides Fr. var. mandshuricum (Maxim) Kitag (Chinese wild ginger) is an important medicinal herb. Essential oil extracted from its roots is the key ingredient and is mainly composed of phenylpropanoid compounds. As a skiophyte plant, light is a crucial factor for A. heterotropoides var. mandshuricum growth and metabolism. To investigate the effects of light irradiation on the essential oil biosynthesis in A. heterotropoides var. mandshuricum, the plants were cultivated in four light irradiation treatments (100, 50, 24 and 12% full sunlight). The photosynthetic capacity, essential oil content and composition, activities of several enzymes and levels of some secondary metabolites involved in the shikimic acid and cinnamic acid pathways were analyzed. The leaf mass per area, average diurnal net photosynthetic rate, and the essential oil content increased significantly with increasing light intensity. Phenylalanine, cinnamic acid, and p-coumaric acid in the cinnamic acid pathway were at their highest levels in plants cultivated in 100% full sunlight. The highest content of shikimic acid in the shikimic acid pathway was obtained in plants grown in 50% sunlight transmittance. The activity of the enzymes 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase, phenylalanine ammonia lyase, cinnamate-4-hydroxylase and 4-coumarate:CoA ligase increased proportionally with light intensity. Overall, we conclude that high light irradiation promotes high net photosynthetic rate, high activity of enzymes and high amounts of phenylpropanoid precursor metabolites leading to significant biosynthesis of essential oil in A. heterotropoides var. mandshuricum.

Colored shade nets induced changes in growth, anatomy and essential oil of Pogostemon cablin.

The purpose of this investigation was to determine the influence of colored shade nets on the growth, anatomy and essential oil content, yield and chemical composition of Pogostemon cablin. The plants were cultivated under full sunlight, black, blue and red nets. The harvesting was performed 5 months after planting and it was followed by the analysis of plant growth parameters, leaf anatomy, essential oil content, yield and chemical composition. The plants grown under red net have produced more leaf, shoot, total dry weight and leaf area. Plants cultivated under colored nets showed differences in morphological features. Plants maintained under red net had a higher leaf blade thickness and polar and equatorial diameter of the stomata ratio. Additionally, higher yield of essential oil in the leaves was observed under red and blue colored shade net. The essential oil of the plants grown under red net showed the highest relative percentage of patchoulol (66.84%). Therefore, it is possible using colored shade nets to manipulate P. cablin growth, as well as its essential oil production with several chemical compositions. The analyses of principal components allowed observing that pogostol has negative correlation with α-guaiene and α-bulnesene. There was difference in total dry weight and patchoulol content when the patchouli is cultured under the red colored shade nets.

Induction of stress volatiles and changes in essential oil content and composition upon microwave exposure in the aromatic plant Ocimum basilicum.

Exposure to sustained low intensity microwaves can constitute a stress for the plants, but its effects on plant secondary chemistry are poorly known. We studied the influence of GSM and WLAN-frequency microwaves on emissions of volatile organic compounds and content of essential oil in the aromatic plant Ocimum basilicum L. hypothesizing that microwave exposure leads to enhanced emissions of stress volatiles and overall greater investment in secondary compounds. Compared to the control plants, microwave irradiation led to decreased emissions of ß-pinene, α-phellandrene, bornyl acetate, ß-myrcene, α-caryophyllene and benzaldehyde, but increased emissions of eucalyptol, estragole, caryophyllene oxide, and α-bergamotene. The highest increase in emission, 21 times greater compared to control, was observed for caryophyllene oxide. The irradiation resulted in increases in the essential oil content, except for the content of phytol which decreased by 41% in the case of GSM-frequency, and 82% in the case of WLAN-frequency microwave irradiation. The strongest increase in response to WLAN irradiation, >17 times greater, was observed for hexadecane and octane contents. Comparisons of volatile compositions by multivariate analyses demonstrated a clear separation of different irradiance treatments, and according to the changes in the volatile emissions, the WLAN-frequency irradiation represented a more severe stress than the GSM-frequency irradiation. Overall, these results demonstrating important modifications in the emission rates, essential oil content and composition indicate that microwave irradiation influences the quality of herbage of this economically important spice plant.

Characterization of the Aldehydes and Their Transformations Induced by UV Irradiation and Air Exposure of White Guanxi Honey Pummelo (Citrus Grandis (L.) Osbeck) Essential Oil.

Aldehydes are key aroma contributors of citrus essential oils. White Guanxi honey pummelo essential oil (WPEO) was investigated in its aldehyde constituents and their transformations induced by UV irradiation and air exposure by GC-MS, GC-O, and sensory evaluation. Nine aldehydes, i.e., octanal, nonanal, citronellal, decanal, trans-citral, cis-citral, perilla aldehyde, dodecanal, and dodecenal, were detected in WPEO. After treatment, the content of citronellal increased, but the concentrations of other aldehydes decreased. The aliphatic aldehydes were transformed to organic acids. Citral was transformed to neric acid, geranic acid, and cyclocitral. Aldehyde transformation caused a remarkable decrease in the minty, herbaceous, and lemon notes of WPEO. In fresh WPEO, ß-myrcene, d-limonene, octanal, decanal, cis-citral, trans-citral, and dodecenal had the highest odor dilution folds. After the treatment, the dilution folds of decanal, cis-citral, trans-citral, and dodecenal decreased dramatically. This result provides information for the production and storage of aldehyde-containing products.

The role of supplemental ultraviolet-B radiation in altering the metabolite profile, essential oil content and composition, and free radical scavenging activities of Coleus forskohlii, an indigenous medicinal plant.

The effects of supplemental ultraviolet-B (s-UV-B; 3.6 kJ m(-2) day(-1) above ambient) radiation were investigated on plant metabolite profile, essential oil content and composition, and free radical scavenging capacities of methanolic extracts of Coleus forskohlii (an indigenous medicinal plant) grown under field conditions. Essential oil was isolated using hydrodistillation technique while alterations in metabolite profile and oil composition were determined via gas chromatography-mass spectroscopy (GC-MS). Leaf and root methanolic extracts were investigated via various in vitro assays for their DPPH radical-, superoxide radical-, hydrogen peroxide-, hydroxyl radical-, and nitric oxide radical scavenging activities, ferrous ion chelating activity, and reducing power. Phytochemical analysis revealed the presence of alkaloids, anthocyanins, coumarins, flavonoids, glycosides, phenols, saponins, steroids, tannins, and terpenoids. Oil content was found to be reduced (by ∼7 %) in supplemental UV-B (s-UV-B) treated plants; the composition of the plant extracts as well as essential oil was also considerably altered. Methanolic extracts from treated plant organs showed more potency as free radical scavengers (their EC50 values being lower than their respective controls). Anomalies were observed in Fe(2+) chelating activity for both leaves and roots. The present study concludes that s-UV-B adversely affects oil content in C. forskohlii and also alters the composition and contents of metabolites in both plant extracts and oil. The results also denote that s-UV-B treated plant organs might be more effective in safeguarding against oxidative stress, though further studies are required to authenticate these findings.

Cinnamon oil nanoemulsion formulation by ultrasonic emulsification: investigation of its bactericidal activity.

Cinnamon oil (extracted from Cinnamomum zeylanicum) nanoemulsion was formulated using Tween 80 and water by ultrasonic emulsification. Process of nanoemulsion formulation was optimized for parameters such as surfactant concentration, oil-surfactant mixing ratio and emulsification time. Surfactant concentration was found to be inversely related to droplet size and directly related to stability. Increase in emulsification time resulted in decrease in droplet diameter. Stable cinnamon oil formulation (CF3) having droplet diameter of 65 nm was formulated after sonication for 30 min. Formulated nanoemulsion was evaluated for bactericidal efficacy against Bacillus cereus. Time and concentration dependent killing of B. cereus cells was observed upon treatment with nanoemulsion. Even at a higher dilution of CF3, significant reduction in bacterial population was observed. Alteration in membrane permeability of interacted samples was suggested by quantifying the release of UV absorbing materials. Bacterial staining with acridine orange/ethidium bromide supported kinetics of killing data and also substantiated the above findings of alteration in membrane permeability. FTIR illustrated disappearance of peak corresponding phosphate vibration at 1078 cm(-1) and 536 cm(-1), and peak associated with vibration of acyl chains of lipid at 2852 cm(-1) was shifted to 2854 cm(-1) which suggested deformation of membrane phospholipids in nanoemulsion treated cells. SEM observations demonstrated membrane distortion leading to cell lysis. These results propose the potential use of cinnamon oil nanoemulsion for preservation of minimally processed food.

Chemical composition and antioxidant properties of γ-irradiated Iranian Zataria multiflora extracts.

CONTEXT: Irradiation is the process of exposing food such as herbal plant to ionizing radiation to destroy microorganisms. Zataria multiflora Boiss (Lamiaceae), known as Avishan-e-Shirazi in Persian, is a thyme-like plant that grows naturally in central and southern parts of Iran and is used in traditional folk medicine. OBJECTIVE: In this study, the effects of γ-radiation on chemical composition and antioxidant properties of Z. multiflora were investigated. MATERIALS AND METHODS: The plants were first irradiated with Co60 source (0, 10, and 25 kGy) and then subjected to Clevenger extraction to obtain essential oils. The composition of the oil was analyzed by a gas chromatography and compared with samples pretreated under different conditions. In parallel, the hydroalcoholic extract was prepared and used for measuring flavonoid content. Thereafter, the free-radical scavenging and antioxidant properties of essential oils and hydroalcoholic extract were examined. RESULTS: Despite the minor change in the individual oil constituents, the total percentage of the main components remained unaffected before and after irradiation (~95%). In addition, the total flavonoid content of hydroalcoholic extract was also unchanged due to irradiation (~32 mg QE/g extract). The high radical scavenging activity of the oil (~67%) and hydroalcoholic extract (~71%), in addition, the antioxidant properties of the oil (~91%) and hydroalcoholic extract (~95%), were unaffected after irradiation. DISCUSSION AND CONCLUSIONS: These findings may suggest the sustainability of Z. multiflora extract properties pretreated with γ-radiation. With a view to its antioxidant applications, resistance of Z. multiflora and its properties against radiation effects are promising findings.

In vitro photo-induced cytotoxic activity of Citrus bergamia and C. medica L. cv. Diamante peel essential oils and identified active coumarins.

CONTEXT: The search for innovative therapeutic approaches is gaining more interest in clinical oncology. OBJECTIVE: In the present investigation we reported the chemical profile and the photo-induced cytotoxic activity of two endemic Calabrian Citrus species (Rutaceae): Citrus bergamia Risso & Poit. and Citrus medica L. cv. Diamante. MATERIALS AND METHODS: Essential oils were obtained by hydrodistillation and analyzed by GC and GC/MS. In order to evaluate the cytotoxic activity two melanoma models, such as amelanotic melanoma C32 and malignant melanoma A375, were used. RESULTS: The essential oil of C. bergamia was characterized by limonene, linalyl acetate, gamma-terpinene, linalool and beta-pinene as major components. The most abundant compounds of C. medica cv. Diamante oil were limonene, gamma-terpinene, citral, geranial, beta-pinene and alpha-pinene. Two coumarins, bergapten and citropten, were also identified in C. bergamia and C. medica cv. Diamante, respectively and tested for biological activity. Both C. bergamia and C. medica cv. Diamante oils exhibited a selective interesting activity against the A375 cell line with IC(50) values of 79.3 and 89.1 microg/mL, respectively, after 100 min exposure to UV irradiation. The strong antiproliferative activity demonstrated with bergapten (IC(50) value of 71.3 microg/mL after 20 min of irradiation) was not found with citropten. DISCUSSION AND CONCLUSION: Our study suggested that UV irradiation is effective in activating essential oils and in particular bergapten. This phototoxicity may be considered as a treatment option in some cases of lentigo maligna or lentigo maligna melanoma.

Hepatoprotective effects of gamma-irradiated caraway essential oils in experimental sepsis.

Irradiation is an important method of processing herbal drugs, while our understanding of the effects of gamma-irradiation on pharmacological properties of seed products such as caraway essential oils is however still very limited. In this study, caraway seeds were irradiated at dose levels of 0, 10 and 25kGy. After extracting the essential oils, the effects of fresh and gamma-irradiated caraway oils (100mg/kg b.w) on preventing septic-related oxidative liver injury induced by cecal ligation and puncture (CLP) model were investigated by measuring oxidative stress parameters in the liver. CLP operation caused a marked increase in myeloperoxidase (MPO) activity which was readily reversed in rats treated with fresh and irradiated caraway oils. Likewise, thiobarbituric acid reactive substances (TBARS) level in the liver was compensated in rats treated with the fresh and irradiated caraway oils. Moreover, liver GSH which was initially depleted due to CLP was recovered by essential oil treatments. The protective role of oils was further confirmed by showing that liver function tests (ALT/AST) as well as histopathological changes following CLP operation were recovered in rats treated with oils from either fresh or irradiated caraway seeds. These data may suggest that gamma-irradiation to caraway seeds at 10 and 25kGy has no influence on the antioxidative properties of caraway essential oils.

Effect of gamma and e-beam radiation on the essential oils of Thymus vulgaris thymoliferum, Eucalyptus radiata, and Lavandula angustifolia.

The microbiological contamination of raw plant materials is common and may be adequately reduced by radiation processing. This study evaluated the effects of gamma- and e-beam ionizing radiations (25 kGy) on three plants used as food or as medicinal products (Thymus vulgaris L., Eucalyptus radiata D.C., and Lavandula angustifolia Mill.) as well as their effects on extracted or commercial essential oils and pure standard samples. Comparison between irradiated and nonirradiated samples was performed by GC/FID and GC/MS. At the studied doses, gamma and e-beam ionizing radiation did not induce any detectable qualitative or quantitative significant changes in the contents and yields of essential oils immediately after ionizing radiation of plants or commercial essential oils and standards. As the maximum dose tested (25 kGy) is a sterilizing dose (much higher than doses used for decontamination of vegetable drugs), it is likely that even decontamination with lower doses will not modify yields or composition of essential oils of these three plants.

UV-B is required for normal development of oil glands in Ocimum basilicum L. (sweet basil).

Plants of Ocimum basilicum L. grown under glass were exposed to short treatments with supplementary UV-B. The effect of UV-B on volatile essential oil content was analysed and compared with morphological effects on the peltate and capitate glandular trichomes. In the absence of UV-B, both peltate and capitate glands were incompletely developed in both mature and developing leaves, the oil sacs being wrinkled and only partially filled. UV-B was found to have two main effects on the glandular trichomes. During the first 4 d of treatment, both peltate and capitate glands filled and their morphology reflected their 'normal' mature development as reported in the literature. During the following days there was a large increase in the number of broken oil sacs among the peltate glands as the mature glands broke open, releasing volatiles. Neither the number of glands nor the qualitative or quantitative composition of the volatiles was affected by UV-B. There seems to be a requirement for UV-B for the filling of the glandular trichomes of basil.

Influence of gamma-irradiation and microwaves on the antioxidant property of some essential oils.

The antioxidant property of anise, caraway, cumin and fennel essential oils extracted from untreated, gamma-irradiated and microwaved fruits against sunflower oil oxidative rancidity was evaluated. The fruits were exposed to gamma-irradiation at 10 KGy and to microwaves at low oven power setting for 1 min. The essential oils were added individually (200 ppm) to sunflower oil and the rate of oil oxidation was followed by determining the peroxide value during storage at room temperature. The irradiated and microwaved essential oils exhibited an antioxidant activity and was superior to that of sunflower oil catalysed by a mixture of BHT + BHA (200 ppm) in most cases. The present data show that gamma-irradiation and microwave treatments did not affect the antioxidant property of the essential oils under study. In addition the essential oils extracted from the gamma-irradiated fruits were more effective as an antioxidant in sunflower oil than those produced from microwaved fruits.

The effects of gamma-irradiation on some pure aroma compounds of spices.

Chemical changes due to gamma-irradiation were investigated in 11 pure aroma compounds typically found in spices. Samples were irradiated with doses of 0, 10, and 50 kGy. The irradiation source was 60Co. Irradiated samples and unirradiated controls were analyzed by gas chromatography alone and coupled with mass spectrometry. Significant changes due to the irradiation occurred only in (+/-)-linalool and alpha-terpineol, which are typical components of, for example, coriander and bay leaf. Their total amounts decreased by about 4-13% and some minor peaks increased as a result of irradiation. However, different results may be obtained when the spices themselves are irradiated.

The influence of radapertization upon some sensory properties of black pepper.

Radapertization may be used as an effective method for sterilization of spices. A possible influence of irradiation upon the sensoric qualities should be considered in this context. Therefore the objectives of the work were: --to determine the influence of the irradiation of pepper upon its sensoric quality --to determine the qualitative changes of volatile chemical substances under the influence of the irradiation of pepper --to determine correlation between changes resulting from irradiation and the sensoric quality of pepper. The spice was irradiated with doses of 10, 20, 40 and 60 kGy. Thresholds of recognition and difference were determined. It was established that the quantity of the dose is related to the values of thresholds mentioned above. The larger the dose was the less intensive the flavour became and the higher the thresholds. These interdependences were written in a form of mathematical formulas. A significant influence of irradiation upon the decrease of the quantity of essential oils and carbohydrates was noticed. These relations were also presented in a form of mathematical formulas. The work provides evidence for the fact that there is a relation between the results of sensoric assessments and the chemical characteristics (the content of volatile carbonyl, sulphuric and nitrogenic substances). Correlation between these factors was determined on the basic statistical analysis of the results obtained during our investigations.