Composition,antibacterial, antioxidant and antiproliferative activities of essential oils from three Origanum species growing wild in Lebanon and Greece

The essential oils from Origanum dictamnus, Origanum libanoticum and Origanum microphyllum were analysed by GC-MS, finding carvacrol, p-cymene, linalool, γ-terpinene and terpinen-4-ol as major components. The antioxidant activity by the DPPH and FRAP tests and the antiproliferative activity against two human cancer cell lines, LoVo and HepG2, were investigated, showing that the essential oil of O. dictamnus was statistically the most inhibitory on both the cell lines, while all the oils exerted a weak antioxidant activity. Furthermore, the samples were tested against 10 Gram-negative and Gram-positive bacteria; all the oils were active on Gram-positive bacteria but O. dictamnus essential oil was the most effective (MIC = 25–50 μg/mL), showing also a good activity against the Gram-negative Escherichia coli (MIC = 50 μg/mL). Data suggest that these essential oils and particularly O. dictamnus oil could be used as valuable new flavours with functional properties for food or nutraceutical products.     

Multiple analytical approaches for the organic and inorganic characterization of Origanum vulgare L. samples

Origanum vulgare L. samples, marketed in different geographic locations, were characterized by their organic and inorganic chemical composition. A total of 35 commercial samples were collected from various sites and analyzed to determine the qualitative and quantitative profile of essential oils, phenolic compounds and some inorganic elements. The variation in the content and composition of the essential oil was assessed by GC and GC-MS analyses, the phenolic fraction was investigated by UPLC^®/PDA, and the inorganic elements were determined by ICP-MS. The Principal Component Analysis (PCA) was applied with the aim to sort out the Origanum vulgare L. samples with different composition according to the different belonging origins. The results showed appreciable qualitative and quantitative differences among samples from different geographic origin.     

Characterization of Spanish unifloral honeys by solid phase microextraction and gas chromatography-mass spectrometry

Volatile compounds have been investigated in unifloral honeys of the most popular types in Spain. A total of 21 eucalyptus samples, 35 rosemary samples, 33 heather samples, and 15 citrus samples were collected in the course of 3 years in different Spanish regions. Samples were analyzed by SPME followed by GC-MS. About 83 compounds were identified; the concentrations of 33 of them were selected to be processed by multivariate analysis. Discriminant analysis allowed correct assignment of most samples: 94% citrus, 92% eucalyptus, 84% heather, and 84% rosemary. As pure reference honeys are not available, a two step data analysis is proposed, selecting the samples most clearly classified in discriminant analysis as "reference samples" to be used in multiple regression to estimate the most representative compounds for each honey type and considering the rest of the samples as of mixed origin.     

Antioxidant capacity and total phenolic contents of oregano (Origanum vulgare), lavender (Lavandula angustifolia) and lemon balm (Melissa officinalis) from Romania

The study reported here presents a comparative screening of three medicinal plants including oregano (Origanum vulgare L.), lavender (Lavandula angustifolia) and lemon balm (Melissa officinalis) having the same geographical origin, the Southeast region of Romania, and growing in the same natural conditions. The contents of total phenolics and total flavonoids for the extracts of these were determined. Furthermore, the total antioxidant capacity was also evaluated. It was found that Origanum vulgare and Melissa officinalis extracts present the most effective antioxidant capacity in scavenging DPPH radicals, while Lavandula angustifolia is less active. High performance liquid chromatography-mass spectrometry analysis was used to identify the components of extracts. Major phenolic acids identified in the analysed species were ferulic, rosmarinic, p-coumaric and caffeic, while predominant flavonoids were quercetin, apigenin kaempherol, which were present as glucosides.     

Optimization of carvacrol,rosmarinic, oleanolic and ursolic acid extraction from oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.)

The aim of our study was to increase the extraction efficiency of carvacrol, rosmarinic, oleanolic and ursolic acid from the different species of oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.). Various extraction methods (ultrasound-assisted, heat-reflux, continuous stirring, maceration, percolation) and extraction conditions (different solvent, material:solvent ratio, extraction temperature, extraction time) were used, and the active substances were determined by HPLC. The lowest content of carvacrol, rosmarinic, oleanolic and ursolic acid was obtained by percolation. During heat-reflux extraction, the content of active substances depended on the solvent used: ethanol/non-aqueous solvent (glycerol or propylene glycol) mixture was more effective compared with ethanol alone. The results showed that for each species of oregano the most optimal extraction method should be selected to maximize the content of biologically active substances in the extracts.     

Comparison of the oxidative stability of soybean and sunflower oils enriched with herbal plant extracts

The present study was conducted to determine and compare the oxidative stability of soybean and sunflower oils using differential scanning calorimetry (DSC). These edible oils were enriched with marjoram (Origanum majorana L.), thyme (Thymus vulgaris L.), and oregano (Origanum vulgare L.) extracts at three different concentrations and synthetic antioxidant (BHA). The fatty acid composition of studied oils was determined by gas chromatography mass spectrometry to evaluate the content of unsaturated fatty acids that are sensitive to oxidation process. Oil samples were heated in the DSC at different heating rates (4.0, 7.5, 10.0, 12.5, and 15.0 °C min^?1) and oxidation kinetic parameters (activation energy, pre-exponential factor, and oxidation rate constant) were calculated. The results showed that the oxidative stability of sunflower oil samples enriched with oregano extracts and soybean oil supplemented with thyme extracts was improved compared to samples without the addition of herbal plant extracts and the synthetic antioxidant.     

High resolution capillary GC–MS analysis of low molecular weight organic compounds in municipal wastewater

High resolution capillary gas chromatography–mass spectrometry has been used in combination with various sample preparation methods to analyze a wide range of low molecular weight organic contaminants in municipal wastewaters. Volatile organic compounds were extracted using a purge and trap concentrator connected directly to a wide bore DB-624 capillary column. Because of the high organic load in wastewater samples, this method was preferred to the more sensitive closed loop stripping technique which suffered from competition effects and insufficient adsorbent capacity. Volatile and semi-volatile base/neutral compounds and acids were extracted with dichloromethane at pH 11 and pH 2, respectively, or were recovered using a simultaneous distillation-extraction procedure. Acidic compounds were subsequently derivatized using diazomethane methylation. Both fractions were separated with a 30m DB-5ms capillary column. Specific ionic substances (LAS, NTA, EDTA) were isolated by solid-phase extraction then derivatized with diazomethane (LAS) or acetyl chloride in n-butanol (NTA, EDTA) before GC–MS analysis. This array of techniques enabled the identification of more than 150 organic contaminants in the influent of a municipal wastewater treatment plant in northern suburban Paris. The individual concentrations of these contaminants ranged from < 0.1 μg/l to 5.7 mg/l.     

Superheated water extraction of essential oils of Origanum micranthum

Superheated water extraction is used to extract essential oil of leaves of Origanum micranthum. The effect of different temperatures on the essential oil profile and rate of extraction as a function of time is investigated. The components of essential oil of Origanum micranthum are removed from the aqueous extract by C18 solid-phase extraction. The identification of components is carried out using comprehensive gas chromatography-time of flight-mass spectrometry. The number of extracted components is almost the same; however, the concentrations change with changing temperature. The highest yield (0.64%) is found at a temperature of 150 degrees C, 2 mL/min and 60 bar for 30 min. The increasing temperature from 100 degrees C to 175 degrees C increased the rate of extraction of six selected components of essential oil of Origanum micranthum. cis-Sabinenehydrate exhibits the fastest rate of extraction at all temperatures studied. Some degradation products are observed at a temperature of 175 degrees C.     

Trace-level determination of polar flavour compounds in butter by solid-phase extraction and gas chromatography-mass spectrometry.

Volatile compounds are responsible for the aromas of butter. A simple technique for the determination of these components is described which is based on solid-phase extraction (SPE) after melting of the butter and separation of the aqueous phase from the fat. Volatile flavours present in the water fraction are collected by off-line SPE on cartidges packed with a copolymer sorbent. After desorption with 500 microliters of methyl acetate, 1-microliter aliquots are quantified and/or identified by gas chromatography-mass spectrometry. The procedure was tested with respect to recovery, linearity and limit of detection in real-life samples using five polar model analytes. It allows the characterisation of polar flavour compounds in butter prior to and after heat treatment at 170 degrees C. From the five model compounds, vanillin, traces of diacetyl and maltol were found to be present in the butter samples. After heat treatment 500-1000-fold increased concentration of maltol, and substantial amounts of furaneol were detected.     

Analysis of volatile components of Lavandula luisieri L. by direct thermal desorption-gas chromatography-mass spectrometry

Volatile components of wild samples of Lavandula luisieri collected in Central and Southern Spain have been analyzed by direct thermal desorption coupled to gas chromatography-mass spectrometry (DTD-GC-MS). This method requires only 10-20 mg of dry sample, allowing to obtain qualitative and quantitative results from different plant parts such as flowers and leaves. Average volatile yield calculated from 51 individual plants was higher for leaves (9.7mg g(-1)) than for flowers (2.9mg g(-1)). Samples presented a high variation in their yield and composition. Major components were camphor and 1,8-cineole (up to 80.9 and 76.7% in leaves; 87.8 and 85.2% in flowers, respectively); however, these compounds were not detected in several samples. Other major component (up to 60% in flowers and leaves) was 2,3,5,5-tetramethyl-4-methylene-2-cyclopenten-1-one. Multivariate analysis was applied to quantitative data from nine selected compounds in order to show the presence of several patterns in plant composition which were only partially related to the site of collection.     

Polyphenol composition and antioxidant activity of selected medicinal herbs

The aim of this study was to characterize aqueous and alcoholic extracts [30%, 50% and 70% (w/v)] obtained from medicinal herbs (Calendula officinalis, Hypericum perforatum, Galium verum, and Origanum vulgare) used in traditional medicine from our country. Samples were examined for total and individual content of phenolics and antioxidant activities. The highest content of total polyphenols (9.9 ± 0.02 mg gallic acid equivalents (GAE) L^–1 extract) and antioxidant activities expressed as Trolox Equivalent Antioxidant Capacity [307.51 TEAC mmol g^–1DW by the ABTS (2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) method and 20.90 TEAC mmol g^–1 DW by the DPPH (1,1-diphenyl-2-picrylhydrazyl) method] was found in Origanum vulgare (50%) extract. Polyphenolic compounds were quantified using RP-HPLC.     

Determination of phenolic compounds in medicinal plants from the Lamiaceae family

A procedure for the determination of Phenolic compounds in extracts from the medicinal plants of the Lamiaceae family—garden sage (Salvia officinalis L.), creeping thyme (Thymus serpyllum L.), wild marjoram (Origanum vulgare L.), and common balm (Melissa officinalis L.)—obtained under different extraction conditions was developed. The identification of the extracted compounds was performed and their qualitative and quantitative composition was established by HPLC with diode array and mass-spectrometric detection with consideration for the obtained characteristics of the standard samples of individual components. The test samples of medicinal herbs contained caffeic acid (0.19–0.62 mg/g) and rosmaric acid (4–23 mg/g); the highest rosmaric acid content (23 mg/g) was found in wild marjoram, and the lowest content (4 mg/g), in creeping thyme. The extracts of wild marjoram contained the greatest amounts of Phenolic compounds; rosmaric acid and luteolin-7-O-β-D-glucuronide were major components, whereas protocatechuic, 3-O-caffeoylquinic, and caffeic acids were minor components.     

Investigation of bacterial viability from incubated saliva by application of flow cytometry and hyphenated separation techniques

The aim of the study was determination of bacterial viability in saliva samples and finding a correlation between microbiological and volatile profiles of saliva depending on incubation time. Bacteria colonizing healthy oral cavities were also identified. Twelve healthy adults donated unstimulated saliva samples. Flow cytometry, optical density measurements and colony‐forming unit (CFU) counting method were employed for analyses of native and inoculated saliva after 0, 1, 2, 24, and 48 h of incubation. Volatile profiles were acquired using headspace‐solid phase microextraction‐gas chromatography/mass spectrometry (HS‐SPME‐GC/MS). Oral bacteria were the most viable within 2 h after collection of saliva. Extension of incubation time to 48 h caused considerable decrease in live bacteria counts and sharp increase in dead bacteria counts. The most prevalent strain was Sphingomonas paucimobilis (26.67%). The number of volatiles raised from 5 to 27 with incubation time and most of them were putrefaction products, such as methanethiol, indole and pyrrole. HS‐SPME‐GC/MS method is insufficient for volatile profiling of “fresh” saliva and should be directed rather to investigation of bacterial metabolites.     

Chemotherapy control by breath profile with application of SPME‐GC/MS method

Chemotherapy used as a treatment against lung cancer has influence on metabolic processes occurring in healthy cells. The changes of biochemical pathways proceeded inside cells might be observed in expired air. In the experiment, breath analysis was carried out before and after anticancer therapy. Expired air samples were collected from 22 patients with a biopsy confirmed lung cancer. Volatile organic compounds present in breath were analyzed by gas chromatography/mass spectrometry. For enrichment of analytes solid‐phase microextraction technique was applied. Eight fibers covered by different sorbents were tested. Carboxen‐polydimethylsiloxane fiber revealed the highest extraction efficiency in relation to analytes in breath. The data showed that cytostatic drugs increase the concentration of acetone and isoprene in the breath collected after chemotherapy. Volatile metabolites of administrated drugs were not identified in expired air.     

Headspace solid-phase microextraction-gas chromatography-mass spectrometry applied to quality control in multilayer-packaging manufacture

A method based on headspace solid-phase microextraction-gas chromatography-mass spectrometry is proposed for the quality control of multilayer packaging and its manufacturing process. Volatile organic compounds (VOCs) are produced in the manufacturing process of the packaging. They can cause organoleptic problems or modify the properties of the packaging depending on the nature and the amount of the VOCs formed. The quantification using packaging samples with a known VOC concentration for the calibration is proposed in order to reduce the analysis time, and the method is validated using a statistical test. Finally, the method is applied to the determination of odour-responsible compounds in multilayer packaging samples obtained under different extrusion-coating conditions, i.e. type of extruder, type of polymer and extrusion speed.     

Simultaneous determination of volatile aromatic and halogenated hydrocarbons in water and soil by dual-channel ECD/PID equilibrium headspace analysis

Volatile aromatic and halogenated hydrocarbons are determined in water and soil samples by equilibrium headspace gas chromatography. Simultaneous analysis is carried out in a dual column/dual detector arrangement with ECD and PID. Their determination in contaminated soil is performed with the same instrumental configuration. However, in contrast to water analysis, an increased sample temperature was found necessary for highly adsorptive soil specimens. At sample temperatures of 95 °C, recoveries near 100% have been found. Both adsorption and desorption processes were found to be strongly time-dependent.     

固相萃取光度法测定烟草中的挥发酚

研究了用Waters Sep-Park-C18固相萃取小柱萃取测定烟草样品中的挥发酚的方法。用自动水蒸汽蒸馏仪蒸馏出烟草样品中的挥发酚，4-氨基安替比林显色，显色产物可用Waters Sep-Park-C18固相萃取小柱萃取，以乙醇洗脱后用分光光度法测定，该方法可用于烟草样品中挥发酚的测定。     

Contrasting Volatilomes of Livestock Dung Drive Preference of the Dung Beetle Bubas bison (Coleoptera: Scarabaeidae)

Volatile cues can play a significant role in the location and discrimination of food resources by insects. Dung beetles have been reported to discriminate among dung types produced by different species, thereby exhibiting behavioral preferences. However, the role of volatile organic compounds (VOCs) in dung localization and preference remains largely unexplored in dung beetles. Here we performed several studies: firstly, cage olfactometer bioassays were performed to evaluate the behavioral responses of Bubas bison (Coleoptera: Scarabaeidae) to VOCs emanating from fresh horse, sheep, and cattle dung; secondly, concurrent volatilome analysis was performed to characterize volatilomes of these dung types. Bubas bison adults exhibited greater attraction to horse dung and less attraction to cattle dung, and they preferred dung from horses fed a pasture-based diet over dung from those fed lucerne hay. Volatilomes of the corresponding dung samples from each livestock species contained a diverse group of alkanes, alkenes, alkynes, alcohols, aldehydes, ketones, esters, phenols, and sulfurous compounds, but the composition and abundance of annotated VOCs varied with dung type and livestock diet. The volatilome of horse dung was the most chemically diverse. Results from a third study evaluating electroantennogram response and supplementary olfactometry provided strong evidence that indole, butyric acid, butanone, p-cresol, skatole, and phenol, as well as toluene, are involved in the attraction of B. bison to dung, with a mixture of these components significantly more attractive than individual constituents.     

Automatic thermal desorption in GC analysis of dairy product volatiles

Summary Automatic thermal desorption (ATD) has been evaluated for the analysis of volatile components from dairy products. It has resulted in a fast and reproducible method which needs only a low amount of sample. Volatile components from milk are collected by off-line purging and trapped on cartridges packed which a suitable adsorbent. Cheese samples are submitted to the same process or directly introduced into the cartridges. The desorption step that follows is carried out automatically and on-line with the chromatographic analysis. Repeatability and sensitivity were satisfactory for both types of products.     

Ion mobility spectrometry of volatile compounds from Iberian pig fat for fast feeding regime authentication

Characteristic ion mobility spectra for volatile compounds present in fat were used to authenticate the feeding regime of Iberian pigs. Volatile compounds were obtained by heating the solid samples at 150 degrees C for 40 min. This produced a headspace that was introduced in the spectrometer ionization chamber by means of a highly purified nitrogen stream. The spectra thus, obtained for the fat samples were processed chemometrically in order to assess their usefulness for discriminating meat from free-range pigs fed on pasture and acorns and confined pigs fed with commercial feed including high-oleic acid products. Principal component analysis was used as both an exploratory tool and a variable reduction method, and linear discriminant analysis was employed to classify 65 subcutaneous fat samples according to pig feeding regime. Only 2.3% of the samples from pigs reared in confinement were misclassified. 95.5% of the free-range samples were correctly predicted.