A qHNMR method for simultaneous quantification of terpenoids from Ferula ovina (Boiss.) Boiss roots.

Ferula ovina (Boiss.) Boiss is one of the most important endemic medicinal plants in Iran, which has three main terpenoid compounds including ferutinin, stylosin and tschimgine. Ferutinin is the strongest natural phytoestrogen that has agonistic activity on estrogen receptors, particularly α-receptors. To determine the amount of ferutinin in F. ovina roots, we firstly used HPLC-UV method. In the HPLC method, the resolution of ferutinin from the two other compounds, stylosin and tshimgine, was poor. Therefore, we decided to use qHNMR method for simultaneous quantification of ferutinin, stylosin and tshimgine in the plant roots. Quantitative 1H-NMR (qHNMR) was carried out based on the relative ratio of signal integration of each compound [(H-1 for tschimgine (δH 4.94-5.03), OCH3 for stylosin (δH 3.8), and H-9 for ferutinin (δH 5.58)] to certain amount of the internal standard dimethyl sulfone (DMSO2). The qHNMR method showed good precision (intra-day RSD ≤ 2.31%, inter-day RSD ≤ 2.72%), linearity (in the ranges of 1.3-10.41, 1.2-9.7 and 1.1-9.02 mg/mL with correlation coefficients at 0.9991), repeatability (RSD ≤ 2.99%) and stability (RSD ≤ 2.4%) for the quantification of the compounds. This work confirmed that qHNMR represents a feasible alternative to high-performance liquid chromatography based methods for simultaneous quantification of ingredients in plant extracts.

Chemical composition and antioxidant activities of essential oil of Hedychium greenii and Hedychium gracile from India.

The chemical constituents of the essential oils hydrodistilled from rhizome parts of Hedychium greenii W.W. Sm. and Hedychium gracile Roxb, of family Zingiberaceae, growing in India, were analysed for the first time by GC-FID and GC-MS, respectively. A total of 30 and 29 components representing 99.62 and 96.74% of the total oil were identified in the essential oils of H. greenii and H. gracile, respectively. The major components of H. greenii were bornyl acetate (31.32%), α-pinene (14.49%), camphene (12.81%) and limonene (10.55%), whereas H. gracile was dominated by ß-pinene (25.24%), γ-terpinene (24.62%), terpinen-4-ol (14.87%) and 1,8-cineole (7.51%). Essential oils were assessed for antioxidant potential using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging assay. H. greenii oil exhibited stronger antioxidant potential as compared to H. gracile oil and butylated hydroxytoluene (BHT). Thus, H. greenii rhizome oil has the potential to be used as an alternative source of antioxidant.

Essential oil composition of aerial parts from Algerian Anacyclus monanthos subsp. cyrtolepidioides (Pomel) Humphries.

The chemical composition of the essential oil from the aerial parts of Anacyclus monanthos subsp. cyrtolepidioides (Pomel) Humphries (Asteraceae) growing in a semi-arid region of Algeria was investigated for the first time. The essential oil was obtained by hydrodistillation and fully characterized by gas chromatography-mass spectrometry (GC-MS). A total of 97 compounds were identified. The essential oil was found to be rich in trans-chrysanthenyl acetate (9.8 ± 2.0%), (E)-ß-farnesene (7.4 ± 1.5%), germacrene D (6.9 ± 1.3%) and myristicin (4.8 ± 0.8%).

Variation in essential oil composition of Teucrium hircanicum L. from Iran-A rich source of (E)-α-bergamotene.

In present work, the chemical composition of the essential oils obtained from dried flowering aerial parts of Teucrium hircanicum L. (Labiatae) originated from ten wild populations in Iran was analyzed by a GC-FID and GC/MS system. The oil yields varied from 0.04% to 0.1%. A total of thirty-two compounds representing 67.6-97.7% of the oil were identified. The essential oil was found to be rich in sesquiterpene hydrocarpons (E)-α-bergamotene (17.5-86.9%) and (E)-ß-farnesene (0.5-21.4%). Of the total identified compounds, sesquiterpene hydrocarpons (36.1-89.7%) were included the greatest essential oil fraction in all the populations, followed by oxygenated monoterpenes (2.2-21.6%), oxygenated sesquiterpenes (0.0-14.4%) and monoterepene hydrocarbons (0.0-9.5%). Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA) were used to distinguish any geographical variations, indicating that the clustering of populations is related to their geographic origin. According to the GC/MS analysis, two chemotypes consisting of (E)-α-bergamotene and (E)-α-bergamotene-(E)-ß-farnesene were identified in the populations.

Schinus molle essential oil as a potential source of bioactive compounds: antifungal and antibacterial properties.

AIMS: The study was focused on the evaluation of antimicrobial activity in vitro of the essential oil (EO) of leaves from Schinus molle against bacteria and fungi of clinical importance in the search for the discovery of new active compounds. METHODS AND RESULTS: The chemical composition of the S. molle EO was determined by gas chromatography/mass spectrometry and its antimicrobial effect was verified by broth microdilution method. The major compounds found were ß-pinene (25·23%), epi-α-cadinol (21·29%), α-pinene (18·72%), myrcene (11·54%) and sabinene (5·02%). The EO showed significant antifungal activity against Paracoccidioides brasiliensis (39·06 µg ml-1 ), weak action against Cryptococcus neoformans (625 µg ml-1 ) and Trichophyton quinckeanum (625 µg ml-1 ) and was inactive against Candida sp. In the analysis of the antibacterial action, the micro-organisms tested did not show sensitivity. CONCLUSIONS: This study showed a promising result of S. molle volatiles against the fungus P. brasiliensis, which causes paracoccidioidomycosis (PCM), a systemic mycosis of great clinical importance in Latin America. SIGNIFICANCE AND IMPACT OF THE STUDY: The results found here are novel and encourage investigations of the compounds present in this EO, which represents a source of molecules with potential use in the treatment of PCM.

Piper nigrum Oil - Determination of Selected Terpenes for Quality Evaluation.

The growing demand and commercial value of black pepper (Piper nigrum) has resulted in considerable interest in developing suitable and cost-effective methods for chemical characterization and quality evaluation purposes. In the current study, an extensive set of oil samples (n = 23) that were extracted by steam distillation from black pepper seeds was investigated to compare the chemical profiles of samples originating from nine major producing countries, as well as to identify potential chemical markers for quality evaluation. The twenty-two most abundant volatile compounds, mainly terpenes, in these oils were determined by conventional GC/MS analysis. Principal component analysis with this set of data revealed distinct clusters for samples that originated from China and Malaysia. Relatively low concentrations of sabinene (< 0.2%) and high concentrations of 3-carene (10.9 - 21.1%) were observed in these samples, respectively, compared to oil samples from other countries. The enantiomeric distributions of key terpene markers, viz., ß-pinene, sabinene, limonene, and terpinen-4-ol, were determined by chiral GC/MS analysis. Interestingly, for these four monoterpenes, levo-isomers were found to be predominant, emphasizing the highly conserved enzymatic processes occurring in P. nigrum. Moreover, consistent enantiomeric ratios ((-) isomer/(+) isomer) of 92.2 ± 3.0% for ß-pinene, 94.8 ± 2.8% for sabinene, 60.7 ± 1.1% for limonene, and 78.3 ± 1.3% for terpinen-4-ol were observed, independent of geographical location. These results demonstrate the potential of using stereospecific compositions as chiral signatures for establishing the authenticity and quality of black pepper oil.

Simultaneous determination of iprodione, procymidone, and chlorflurenol in lake water and wastewater matrices by GC-MS after multivariate optimization of binary dispersive liquid-liquid microextraction.

This study reports the optimization of a binary dispersive liquid-liquid microextraction method for the determination of iprodione, procymidone, and chlorflurenol by gas chromatography mass spectrometry. The study was aimed at using two extraction solvents to increase the extraction efficiency of all analytes. The binary solvents recorded results higher than the mono-solvents. After examining the effects of main experimental parameters and their interactions by analysis of variance, 200 µL of binary mixture (dichloromethane and 1,2-dichloroethane), 2.5 mL of ethanol, and 15 s vortex were obtained as optimum parameters. The detection and quantification limits calculated for the analytes were found to be between 0.30-1.6 and 1.0-5.3 ng/mL, respectively. Enhancement in detection power calculated as a ratio of the binary extraction detection limit to the detection limit of direct GC-MS analysis was 105-, 214-, and 233-fold for chlorflurenol, iprodione, and procymidone, respectively. In order to check the accuracy of the developed method, recovery study was performed. Water sampled from a lake and two wastewater samples from treatment facilities were spiked at two concentrations, and the percent recovery calculated for the samples ranged between 87 and 116%. These results confirmed the suitability of the method to real samples for accurate determination of the analytes at trace levels.

Jasmonic acid changes the composition of essential oil isolated from narrow-leaved lavender propagated in in vitro cultures.

The aim of the present study was to determine the effect of jasmonic acid added to the culture medium on composition of Lavandula angustifolia essential oils. The chemical composition was determined by gas chromatography coupled to mass detector (GC/MS). The experiment was conducted with the use of MS medium supplemented with increasing concentration of JA (0.2, 0.5, 1, 1.5 mg∙dm-3). It was found that the analysed essential oils varied in terms of chemical composition depending on the content of JA in the medium. All obtained essential oils were characterised by a high content of σ-cadinene (17.06-29.64%), borneol (6.66-17.47%), caryophyllene oxide (8.30-14.01%), τ-cadinol (4.87-9.16%), beta-caryophyllene (3.54-6.57%), 1.8-cineole (1.94-5.87%), ß-pinene (1.48-3.05%), geranyl acetate (0.56-2.14%) and myrtenal (0.65-2.14%).

Attraction behaviour of Anagrus nilaparvatae to remote lemongrass (Cymbopogon distans) oil and its volatile compounds.

Utilisation of Anagrus nilaparvatae is a promising and effective method for planthoppers manipulation. Twenty-seven components of remote lemongrass (Cymbopogon distans) oil were identified by GC/MS and nine volatiles were selected for behavioural experiments. In this study, we noted that the remote lemongrass oil was attractive to female A. nilaparvatae at concentrations of 0.1 and 1 mg/L. α-Pinene, ß-pinene, eucalyptol, carveol and D-carvone attracted female wasps in the dose-dependent bioassays. Blend 1 (a mixture of eucalyptol, D-carvone, carveol, α-pinene, and ß-pinene with ratios of remote lemongrass oil volatiles of 625:80:11:5:3) attracted female wasps at 10 mg/L, while blend 2 (a mixture of the same five volatiles at the same loading ratio) attracted them at 0.1 and 1 mg/L. These results suggested that plant essential oils could be attractants for natural enemies to control pests. The ratios of volatiles in the mixtures affect the attractiveness of the synthetic mixtures.

Phytochemistry and Pharmacology of Ferula hermonis Boiss. - A Review.

Ferula hermonis, a well-known species of the genus Ferula found in Lebanon and Syria, has a brilliant history in traditional medicine as it has been used for the treatment of erectile dysfunction in men and menopausal disturbances in women. Thanks to modern pharmacological and clinical investigations, F. hermonis is a valuable medicinal and condimental plant that may be used for the treatment of impotence and diabetes, the prevention of osteoporosis, and possesses anti-microbial and anti-inflammatory properties. Phytochemical investigations have shown that this plant contains daucane aryl esters such as ferutinin, which has exhibited various biological activities including hypoglycemic and estrogenic activities. Ferutinin is one of the strongest natural phytoestrogen which has agonistic activity on estrogen receptors, particularly α receptor. It seems that ferutinin and its derivatives play an important role in F. hermonis biological activities, mainly the beneficial effects of this plant on impotence, diabetes and osteoporosis. The present review discusses the available data on the active constituents and biological activities of F. hermonis and their possible underlying mechanisms of action.

Habitat-related variation in composition of the essential oil of Seseli rigidum Waldst. & Kit. (Apiaceae).

Plant specialised metabolites like essential oils are highly variable depending on genetic and various ecological factors. The aim of the present work was to characterise essential oils of the species Seseli rigidum Waldst. & Kit. (Apiaceae) in various organs on the individual and populational levels. Geographical variability and the impact of climate and soil type on essential oil composition were also investigated. Individually sampled essential oils of roots, aerial parts and fruits of plants from seven populations were analysed by GC-FID and GC-MS. The investigated populations showed high interpopulational and especially intrapopulational variability of essential oil composition. In regard to the variability of essential oils, different chemotypes were defined. The essential oils of S. rigidum roots represented a falcarinol chemotype, oils of aerial parts constituted an α-pinene or α-pinene/sabinene chemotype and fruit essential oils can be characterised as belonging to a complex sabinene/α-pinene/ß-phellandrene/falcarinol/germacrene B chemotype. At the species level, analysis of variance (ANOVA), principal component analysis (PCA) and canonical discriminant analysis (CDA) showed that the plant part exerted the strongest influence on the composition of essential oils. Climate had a high impact on composition of the essential oils of roots, aerial parts and fruits, while influence of the substrate was less pronounced. The variations in main compounds of essential oils based on climate or substrate were complex and specific to the plant part.

Residual Behaviors of Six Pesticides in Shiitake from Cultivation to Postharvest Drying Process and Risk Assessment.

The dissipation of six pesticides (carbendazim, thiabendazole, procymidone, bifenthrin, λ-cyhalothrin, and ß-cyfluthrin) in shiitakes from cultivation to postharvest drying process was investigated, and the dietary exposure risk was estimated thereafter. The field trial study indicates that the half-lives of carbendazim, thiabendazole, and procymidone were much shorter than those of bifenthrin, λ-cyhalothrin, and ß-cyfluthrin. Furthermore, the effects of two drying processes on the residues and processing factors (PFs) were investigated. The results showed that hot-air drying resulted in higher residues than sunlight exposure drying. Both drying processes led to pesticide residue concentration (with PF > 1), except for thiabendazole upon sunlight exposure treatment. The estimated daily intakes (EDIs) ranged from 0.06% of the acceptable daily intake (ADI) for thiabendazole to 42.43% of the ADI for procymidone. The results show that the six pesticide residues in dried shiitakes are still within acceptable levels for human consumption on the basis of a dietary risk assessment.

Organic aerosol molecular composition and gas-particle partitioning coefficients at a Mediterranean site (Corsica).

Molecular speciation of atmospheric organic matter was investigated during a short summer field campaign performed in a citrus fruit field in northern Corsica (June 2011). Aimed at assessing the performance on the field of newly developed analytical protocols, this work focuses on the molecular composition of both gas and particulate phases and provides an insight into partitioning behavior of the semi-volatile oxygenated fraction. Limonene ozonolysis tracers were specifically searched for, according to gas chromatography-mass spectrometry (GC-MS) data previously recorded for smog chamber experiments. A screening of other oxygenated species present in the field atmosphere was also performed. About sixty polar molecules were positively or tentatively identified in gas and/or particle phases. These molecules comprise a wide range of branched and linear, mono and di-carbonyls (C3-C7), mono and di-carboxylic acids (C3-C18), and compounds bearing up to three functionalities. Among these compounds, some can be specifically attributed to limonene oxidation and others can be related to α- or ß-pinene oxidation. This provides an original snapshot of the organic matter composition at a Mediterranean site in summer. Furthermore, for compounds identified and quantified in both gaseous and particulate phases, an experimental gas/particle partitioning coefficient was determined. Several volatile products, which are not expected in the particulate phase assuming thermodynamic equilibrium, were nonetheless present in significant concentrations. Hypotheses are proposed to explain these observations, such as the possible aerosol viscosity that could hinder the theoretical equilibrium to be rapidly reached.

Chemical composition, toxicity and non-target effects of Pinus kesiya essential oil: An eco-friendly and novel larvicide against malaria, dengue and lymphatic filariasis mosquito vectors.

Mosquitoes (Diptera: Culicidae) are vectors of important parasites and pathogens causing death, poverty and social disability worldwide, with special reference to tropical and subtropical countries. The overuse of synthetic insecticides to control mosquito vectors lead to resistance, adverse environmental effects and high operational costs. Therefore, the development of eco-friendly control tools is an important public health challenge. In this study, the mosquito larvicidal activity of Pinus kesiya leaf essential oil (EO) was evaluated against the malaria vector Anopheles stephensi, the dengue vector Aedes aegypti and the lymphatic filariasis vector Culex quinquefasciatus. The chemical composition of the EO was analyzed by gas chromatography-mass spectroscopy. GC-MS revealed that the P. kesiya EO contained 18 compounds. Major constituents were α-pinene, ß-pinene, myrcene and germacrene D. In acute toxicity assays, the EO showed significant toxicity against early third-stage larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus, with LC50 values of 52, 57, and 62µg/ml, respectively. Notably, the EO was safer towards several aquatic non-target organisms Anisops bouvieri, Diplonychus indicus and Gambusia affinis, with LC50 values ranging from 4135 to 8390µg/ml. Overall, this research adds basic knowledge to develop newer and safer natural larvicides from Pinaceae plants against malaria, dengue and filariasis mosquito vectors.

Essential-Oil Variability in Natural Populations of Pinus mugo Turra from the Julian Alps.

The composition and variability of the terpenes and their derivatives isolated from the needles of a representative pool of 114 adult trees originating from four natural populations of dwarf mountain pine (Pinus mugo Turra) from the Julian Alps were investigated by GC-FID and GC/MS analyses. In total, 54 of the 57 detected essential-oil components were identified. Among the different compound classes present in the essential oils, the chief constituents belonged to the monoterpenes, comprising an average content of 79.67% of the total oil composition (74.80% of monoterpene hydrocarbons and 4.87% of oxygenated monoterpenes). Sesquiterpenes were present in smaller amounts (average content of 19.02%), out of which 16.39% were sesquiterpene hydrocarbons and 2.62% oxygenated sesquiterpenes. The most abundant components in the needle essential oils were the monoterpenes δ-car-3-ene, ß-phellandrene, α-pinene, ß-myrcene, and ß-pinene and the sesquiterpene ß-caryophyllene. From the total data set of 57 detected compounds, 40 were selected for principal-component analysis (PCA), discriminant analysis (DA), and cluster analysis (CA). The overlap tendency of the four populations suggested by PCA, was as well observed by DA. CA also demonstrated similarity among the populations, which was the highest between Populations I and II.

Composition of the Essential Oil of Allium neapolitanum Cirillo Growing Wild in Sicily and its Activity on Microorganisms Affecting Historical Art Crafts.

Essential oil of the aerial parts of Allium neapolitanum Cirillo collected in Sicily were analyzed by gas-chromatography-flame-ionization detection and gas-chromatography-mass spectrometry. Nineteen compounds were identified in the oil and the main components were found to be (E)-chrysanthenyl acetate (28.1%), (Z)-chrysanthenyl acetate (23.8%), (E)-ß-farnesene (9.6%), dimethyl trisulfide (9.6%), camphor (7.4%), methyl allyl disulfide (6.8%) and 1-methyl-3-allyl trisulfide (5.8%). The essential oil showed good antimicrobial activity against 11 strains of test microorganisms, including several species infesting historical material.

Essential Oil of Amomum maximum Roxb. and Its Bioactivities against Two Stored-Product Insects.

Amomum maximum Roxb. is a perennial herb distributed in South China and Southeast Asia. The objective of this work was to analyze the chemical constituents and assess insecticidal and repellent activities of the essential oil from Amomum maximum fruits against Tribolium castaneum (Herbst) and Liposcelis bostrychophila (Badonnel). The essential oil was obtained by hydrodistillation and analyzed by gas chromatography-flame ionization detector and gas chromatography-mass spectrometry. The main components of the essential oil were identified to be ß-pinene (23.39%), ß-caryophyllene (16.43%), α-pinene (7.55%), sylvestrene (6.61%) and ç-cadinene (4.19%). It was found that the essential oil of A. maximum fruits possessed contact and fumigant toxicities against T. castaneum adults (LD50 = 29.57 µg/adult and LC(50) = 23.09 mg/L air, respectively) and showed contact toxicity against L. bostrychophila (LD(50) = 67.46 µg/cm(2)). Repellency of the crude oil was also evaluated. After 2 h treatment, the essential oil possessed 100% repellency at 78.63 nL/cm(2) against T. castaneum and 84% repellency at 63.17 nL/cm(2) against L. bostrychophila. The results indicated that the essential oil of A. maximum fruits had the potential to be developed as a natural insecticide and repellent for control of T. castaneum and L. bostrychophila.

Impact of gas chromatography and mass spectrometry combined with gas chromatography and olfactometry for the sex differentiation of Baccharis articulata by the analysis of volatile compounds.

The Baccharis genus has more than 400 species of aromatic plants. However, only approximately 50 species have been studied in oil composition to date. From these studies, very few take into consideration differences between male and female plants, which is a significant and distinctive factor in Baccharis in the Asteraceae family. Baccharis articulata is a common shrub that grows wild in south Brazil, northern and central Argentina, Bolivia, Paraguay and Uruguay. It is considered to be a medicinal plant and is employed in traditional medicine. We report B. articulata male and female volatile composition obtained by simultaneous distillation-extraction technique and analyzed by gas chromatography with mass spectrometry. Also, an assessment of aromatic differences between volatile extracts was evaluated by gas chromatography with olfactometry. The results show a very similar chemical composition between male and female extracts, with a high proportion of terpene compounds of which ß-pinene, limonene and germacrene D are the main components. Despite the chemical similarity, great differences in aromatic profile were found: male plant samples exhibited the strongest odorants in number and intensity of aromatic attributes. These differences explain field observations which indicate differences between male and female flower aroma, and might be of ecological significance in the attraction of pollinating insects.

A breath fungal secondary metabolite signature to diagnose invasive aspergillosis.

BACKGROUND: Invasive aspergillosis (IA) remains a leading cause of mortality in immunocompromised patients, in part due to the difficulty of diagnosing this infection. METHODS: Using thermal desorption-gas chromatography/mass spectrometry, we characterized the in vitro volatile metabolite profile of Aspergillus fumigatus, the most common cause of IA, and other pathogenic aspergilli. We prospectively collected breath samples from patients with suspected invasive fungal pneumonia from 2011 to 2013, and assessed whether we could discriminate patients with proven or probable IA from patients without aspergillosis, as determined by European Organization for Research and Treatment of Cancer/Mycoses Study Group consensus definitions, by direct detection of fungal volatile metabolites in these breath samples. RESULTS: The monoterpenes camphene, α- and ß-pinene, and limonene, and the sesquiterpene compounds α- and ß-trans-bergamotene were distinctive volatile metabolites of A. fumigatus in vitro, distinguishing it from other pathogenic aspergilli. Of 64 patients with suspected invasive fungal pneumonia based on host risk factors, clinical symptoms, and radiologic findings, 34 were diagnosed with IA, whereas 30 were ultimately diagnosed with other causes of pneumonia, including other invasive mycoses. Detection of α-trans-bergamotene, ß-trans-bergamotene, a ß-vatirenene-like sesquiterpene, or trans-geranylacetone identified IA patients with 94% sensitivity (95% confidence interval [CI], 81%-98%) and 93% specificity (95% CI, 79%-98%). CONCLUSIONS: In patients with suspected fungal pneumonia, an Aspergillus secondary metabolite signature in breath can identify individuals with IA. These results provide proof-of-concept that direct detection of exogenous fungal metabolites in breath can be used as a novel, noninvasive, pathogen-specific approach to identifying the precise microbial cause of pneumonia.

Essential-oil diversity of Salvia tomentosa Mill. in Greece.

Salvia tomentosa essential oils from Greece were studied for the first time here. The oils from five populations growing in Mediterranean pine forests on the island of Thassos (northern Aegean Sea) and from 14 populations situated in deciduous forests in Thrace (northeastern Greek mainland) were investigated. Their essential-oil contents ranged from 1.1 to 3.3% (v/w, based on the dry weight of the plant material). The populations from Thassos had high contents of α-pinene (18.0 ± 2.9%), 1,8-cineole (14.7 ± 3.0%), cis-thujone (14.0 ± 6.9%), and borneol (12.8 ± 2.2%) and smaller amounts of camphene, camphor, and ß-pinene, whereas the populations from Thrace showed high α-pinene (16.7 ± 4.0%), ß-pinene (22.8 ± 4.5%), camphor (18.3 ± 4.3%), and camphene (10.3 ± 2.4%) contents, much lower 1,8-cineole and borneol amounts, while cis-thujone was completely lacking. The comparison of the present results with published data showed that oils having cis-thujone as one of the main compounds were reported for the first time here. Multivariate statistical analyses indicate that the observed essential-oil variation was related to geographical and environmental factors.