Neolignans and polyoxygenated seco-cyclohexenes from the stems and leaves of Piper suipigua Buch.-Ham. ex D. Don.

Five new compounds including, a neolignan, eupomatenoid-19 (1) and four polyoxygenated seco-cyclohexenes, artahongkongenes G-J (2-5), together with fifteen known compounds (6-20) were isolated from the stems and leaves of Piper suipigua Buch.-Ham. ex D. Don. Their structures were determined by spectroscopic evidence (IR, UV, 1H NMR, 13C NMR and 2 D NMR) as well as MS. The absolute configurations of polyoxygenated seco-cyclohexenes 2-8 were identified by NOESY data and by comparison of their experimental and calculated ECD spectral data. Neolignans, eupomatenoid-19 (1) and eupomatenoid-7 (10), displayed cytotoxicity against several cancer cell lines. In addition, eupomatenoid-7 (10) showed antibacterial activity against Bacillus cereus, Bacillus subtilis and Staphylococcus aureus.

Intranasal administration of perillyl alcohol-loaded nanoemulsion and pharmacokinetic study of its metabolite perillic acid in plasma and brain of rats using ultra-performance liquid chromatography/tandem mass spectrometry.

Perillyl alcohol (POH) is a monocyclic terpene that has strong antitumor activity. Brain tumors are particularly difficult to treat with therapeutic agents, and clinical trials have shown their low tolerance through oral administration. We proposed the entrapment of POH into an oil-in-water chitosan nanoemulsion aiming its intranasal administration for brain targeting. An ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the quantitation of total metabolite perillic acid (PA) in plasma and brain of rats. The rat samples containing the metabolite were treated by liquid-liquid extraction with acetonitrile. The mobile phase was 0.1% formic acid in water (solvent A) and 0.1% formic acid in methanol (solvent B), at a flow rate of 0.3 mL min-1 in gradient elution. The chromatography was run for 10 min, and analytical curves were built in acetonitrile, plasma, and brain. The PA was detected in positive ion mode with multiple reaction monitoring. The method has shown high selectivity, sensitivity, and throughput. The low quantification limits of 162, 178, and 121 ng mL-1 for acetonitrile, brain, and plasma, respectively, indicate a good detectability of the method. The repeatability and precision observed were within the limits recommended in the literature. The accuracy of the method was verified through high recovery rates (106-118%). The validated method was successfully applied to the pharmacokinetic study of the metabolite PA after the intranasal administration of free or POH-loaded nanoemulsion in rats. The results showed that chitosan nanoemulsion improved the plasma and brain bioavailability of POH, representing a promising alternative to free POH treatment.

[Determination of Cyclohexene in workplace air by Solvent desorption gas chromatography].

Objective: To establish a solvent desorption gas chromatography method for determination of cyclohexene in workplace air. Methods: Cyclohexene in the air of workplace was collected with carbon tube and desorbed by carbon disulfide. The target toxicant was separated with the GC column and analyzed with FID detector, identified by retention time, and quantified by peak area. Results: The linear range of cyclohexene in the air of workplace was 0.77~4 050.00 µg/ml, with a correlation coefficient of 0.9999. The limit of detection was 0.23 µg/ml. The lower limit of quantification was 0.77 µg/ml. The minimum detectable concentration was 0.15 mg/m(3) under1.5 L sampling volume and 1.0 ml extraction solution volume. The within-run precision of different cyclohexene concentrations was 0.62%~1.9% and the between-run precisions was 1.5%~3.5%; The average extraction efficiency was 96.4%; Penetration capacity (100 mg of carbon tube) was 29.4 mg; The average collection efficiency was 100%; The samples could be stored for 7 days at room temperature. When placed in 4 ℃ refrigerator, the samples could be stored for 14 days. The potential coexistence of cyclohexane, hexane, benzene, toluene and ethylbenzene with cyclohexene in the air did not interfere with the results of determination. Conclusion: This method has high sensitivity, precision, accuracy and lower limit of detection and it is applicable for determination of cyclohexene in workplace air.

Solid phase microextraction and gas chromatography mass spectrometry analysis of Zingiber officinale and Curcuma longa.

SPME analysis of Zingiber officinale Roscoe and Curcuma longa L. were performed by using a DVB/CARB/PDMS fiber. The SPME analysis of Zingiber officinale showed that the main components found were camphene (7.27%), geranial (8.37%), α-zingiberene (14.50%), α-farnesene (9.14%), ß-bisabolene (6.52%), and ß-sesquiphellandrene (9.92%). The SPME analysis of Curcuma longa showed that main components were p-cymene (12.96%) and ar-turmerone (12.08%). Other components were ß-phellandrene (7.86%), terpinolene (6.97%), ar-curcumene (8.53%), α-zingiberene (8.46%), and ß-sesquiphellandrene (7.37%).

Crocus sativus a natural food coloring and flavoring has potent anti-tumor properties.

BACKGROUND: Saffron, the dried stigmas of Crocus sativus L., is a highly valued agricultural product that is used mainly as a food coloring and flavoring agent. Three main secondary metabolites of Crocus sativus including crocin, picrocrocin, and safranal are responsible for the color, the bitter taste and for the odor and aroma, respectively. As a component of traditional medicine, saffron has been utilized as a medicinal herb for treating various ailments including cramps, asthma, liver disease, menstruation disorders, pain, and in the pathogenesis of cancer. HYPOTHESIS/PURPOSE: To summarize the recent published data on the chemo-preventive properties of Crocus sativus in cancer treatment. STUDY DESIGN: We conducted a non-systematic review of the literature. METHODS: A search of English-language literature was performed using Scopus, EMBASE and PubMed. We applied no restriction in time. Articles were searched using the keywords "Lung", "breast", "skin", "prostate", "leukemia", "cancer", "neoplasm", "tumor", "malignancy", "saffron", "crocus sativus", "crocin", "crocetin", "picrocrocin", and "safranal". RESULTS: Saffron has been reported to exert anti-tumor and anti-cancer effects in various types of cancer including lung cancer, breast cancer, leukemia, skin cancer and prostate cancer. This appears to be via various mechanisms including: the induction of apoptosis, arresting cell cycle progression, suppressing expression of matrix metalloproteinase, modulatory effects on some phase II detoxifying enzymes and decreasing expression of inflammatory molecules are potential mechanisms of saffron-induced anticancer effects. CONCLUSION: Saffron possesses potent anti-tumor properties and represents an efficacious and safe treatment.

Dual solid-phase and stir bar sorptive extraction combined with gas chromatography-mass spectrometry analysis provides a suitable tool for assaying limonene-derived mint aroma compounds in red wine.

A novel analytical method was developed for quantitative determination of eight limonene-derived monoterpenes responsible for the mint aroma in red wine. As these aromatic compounds are present at trace levels, a new dual extraction approach was proposed, combining solid-phase extraction (SPE) and stir bar sorptive extraction (SBSE), followed by gas chromatography-mass spectrometry analysis. The various parameters affecting the efficiency of extracting the analytes from wine samples in both the SPE and SBSE steps were first investigated, to determine the best compromise for the simultaneous analysis of the compounds studied. Following preliminary optimization of the dilution factor, phase ratio, and methanol content in the SBSE sample, cartridge sorbent mass, type of solvent, elution volume, and wine sample volume in the pre-concentration SPE step were studied. Highest response values were obtained when a 90 mL wine sample was extracted on a 500 mg SPE C18 cartridge and eluted with 1.5 mL methanol. The wine extract was then diluted in 10 mL water to obtain a final methanol content of 15% before the SBSE step. Good linearity, repeatability, reproducibility, accuracy and the required low detection and quantification limits were obtained under the conditions described, making this SPE-SBSE combination a suitable, powerful tool for routine analysis of the selected limonene-derived mint aroma compounds in large series of wine samples. Finally, the validated method was applied to 15 commercial red Bordeaux wines, aged from 3 to 23 years. Most of the compounds studied, present within the ng.L-1 range, were easily quantified for the first time in wine.

Terpenoid composition of essential oil from a new chemotype of Selinum wallichianum Raizada & Saxena.

Essential oil samples obtained by steam distillation of the whole aerial parts and roots of Selinum wallichianum Raizada & Saxena (syn Selinum tenuifolium) growing wild in the Himalayan region of Uttarakhand, India, were analysed by capillary gas chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS). A total of 24 and 43 constituents representing 97.0 and 95.4% of the oil composition, respectively, were identified. Both the oil samples were constituted mainly of monoterpene hydrocarbons 68.1 and 77.4%, with sabinene 31.0 and 11.5%, ß-phellandrene 18.2 and 34.5%, α-bisabolol 16.0 and 1.8% and α-phellandrene 3.5 and 11.2%, respectively, as a major constituents. Presence of sabinene, ß-phellandrene, α-phellandrene and complete absence of previously reported 3,5-nonadiyne (53.8-90.0%) justifies it to be a new chemotype of S. wallichianum.

Influence of drying methods on chemical compositions, antioxidant and antibacterial activity of essential oil from lemon peel.

The influence of natural, hot-air and infrared drying on chemical composition and bioactivity of lemon peel essential oil are investigated in this study. The results showed that drying resulted in losses or increases of some components or production of some new substances, but the d-limonene (59.52-70.01%) was found as the main component of essential oil. Drying brought about decreases in the yield, antioxidant and antibacterial activity of essential oil. However, the natural drying had little effect, while the hot-air and infrared drying resulted in significant decreases in these parameters, especially at the higher temperature. The yield was the lowest under hot-air drying (60 °C) and decreased by 78%, while infrared drying (60 °C) sample exhibited the lowest antioxidant and antibacterial activities. Infrared drying was easier to lead to the decrease in bioactivity than hot-air drying at the same temperature. These results provided the theoretical basis for drying lemon peel.

Essential oil composition of Erica spiculifolia Salisb - first report.

The essential oil isolated from Erica spiculifolia Salisb. by hydrodistillation was analysed by GC-MS. One hundred compounds representing 92.6% of total oil were identified. Among the detected compounds 38 were monoterpenoids (46.2%), 30 were sesquiterpenoids (31.7%), 2 diterpene (0.4%) and 30 compounds (14.3%) were with various non-terpenoid structures. Oxygenated monoerpenes were by far the main class (45.3%) with α-terpineol (7.5%) endo-borneol (7.2%), pinocarveol (5.9%) and thymol (3.7%) as the major compounds. The predominant components of sesquiterpenes were caryophyllene oxide (5.0%), caryophyllene (4.2%), τ -murrolol (3.5%), spathulenol (2.9) and α-cadinol (2.3%).

Analysis of changes in volatile constituents and expression of genes involved in terpenoid metabolism in oleocellosis peel.

Oleocellosis is a serious physiological disorder in citrus fruit that mainly results in appearance and quality deterioration. It has been well established that the occurrence of oleocellosis is highly correlated with the release of peel oil from citrus fruit, while there is little information on the dynamic changes in the content of the volatile constituents and the expression of genes involved in terpenoid metabolism during oleocellosis development. In the present research, large changes in the volatile profiles and gene expression in terpenoid metabolism were observed in oleocellosis peels compared to healthy ones. Among volatiles, the decreased contents of α-pinene, d-limonene, ß-myrcene, linalool, ß-caryophyllene, α-terpineol, nonanal, neryl acetate and (-)-carvone played a major role in these changes. For gene expressions in terpenoid metabolism, the up-regulated genes aldehyde dehydrogenase (NAD+) (ALDH) and the down-regulated genes ß-caryophyllene synthase 1 (BCS1), α-terpineol synthase 2 (TES2) and myrcene synthase (MS) were the main differences in oleocellosis peels.

Essential oil of Citrus lumia Risso: Phytochemical profile, antioxidant properties and activity on the central nervous system.

The use of essential oils (EOs) is known since long time in traditional medicine and aromatherapy for the management of various oxidative stress-related disorders and has been further increased recently for their neuroprotective and anti-aging potentials as well as for reducing anxiety and stress. The purpose of this work was to evaluate, for the first time, the chemical composition of Citrus lumia Risso EO and its antioxidant, anti-cholinesterase, and neuroactive properties by cell-free and cell-based assays. The EO has shown strong antioxidant and free radical scavenging properties, particularly in hydrogen atom transfer based assays (ß-carotene bleaching and ORAC, IC50 22 µg/mL and 46 µg/mL, respectively), that can be attributed to the high content of monoterpenes, especially d-Limonene (48.905%), and Linalool (18.245%). Furthermore, the EO has shown an interesting anti-acetylcholinesterase activity (IC50 258.25 µg/mL). Data from MTT analysis indicate that the cytotoxicity of EO, evaluated on L929 mouse fibroblasts, is very low, with an IC50 higher than 500 µg/mL at 48 h. Rat neuronal networks subjected to EO showed a concentration-dependent inhibition of spontaneous electrical activity. Results indicate that C. lumia EO could be an important source of natural antioxidants suggesting an important preventive role in the onset of oxidative stress-related pathologies.

The analysis of aroma/flavor compounds in green tea using ice concentration linked with extractive stirrer.

Worldwide, green tea is one of the most popular beverages. It promotes blood circulation, liver function, and lowers the risk of cancer and cardiovascular diseases. This drink is characterized by the distinctive odors and flavors produced by its constituent compounds, with its value predicated on the amount and type of constituents extracted from the tea leaves during brewing. Ice concentration linked with extractive stirrer (ICECLES) is a novel sample preparation technique, especially applicable for the extraction of relatively polar compounds while retaining excellent extraction efficiencies for non-polar compounds. In this study, ICECLES was used to prepare green tea for analysis of aroma/flavor compounds by gas chromatography-mass spectrometry (GC-MS). ICECLES performed very well, revealing 301 constituents as compared to 245 for SBSE (i.e., 56 more constituents were detected via ICECLES). Moreover, ICECLES produced stronger signal to noise ratios for all except 4 of 301 constituents, with a maximum signal enhancement of 19. Of the constituents which were only detectable using ICECLES, some very important aroma/flavor and/or medicinal compounds were easily identified, including furfural, furfural alcohol, maltol, eugenol, 2-methylpyrazine, phenethyl alcohol, 2,6-dimethoxyphenol, and α-terpineol. Overall, we confirmed that ICECLES sample preparation followed by GC-MS consistently allowed more complete green tea aroma/flavor analysis, especially for relatively polar compounds, some of which are critical for flavor quality.

Anti-inflammatory Effect of Essential Oil from Citrus aurantium L. var. amara Engl.

Essential oil has been popularly used as an alternative for the treatment of inflammation. The bioactivities of essential oil from blossoms of Citrus aurantium L. var. amara Engl (CAVAO) showed greater anti-inflammation potential than that of antioxidant, anticancer, and 3T3-L1 proliferation inhibition. CAVAO (250 µg/mL) significantly inhibited production of nitric oxide (NO) (99.54 ± 2.81%), interleukin-6 (IL-6) (98.11 ± 1.62%), tumor necrosis factor-α (TNF-α) (41.84 ± 1.52%), and interleukin-1ß (IL-1ß) (56.09 ± 2.21%) as well as their gene expression level. CAVAO also markedly decreased the expression levels of cyclooxygenase-2 (COX-2) gene and protein. Furthermore, CAVAO inhibited nuclear factor-κB (NF-κB) activation, which was justified by the inhibitory effect on NF-κB nuclear translocation, IκBα phosphorylation and degradation, and phosphorylation-dependent IκB kinase activation in RAW264.7 cells stimulated with lipopolysaccharides. CAVAO also suppressed the phosphorylation of c-Jun N-terminal kinase (JNK) and p38, indicating that mitogen-activated protein kinase (MAPK) signaling pathways were also blocked. The major constituents of CAVAO were characterized as linalool (64.6 ± 0.04%), α-terpineol (7.61 ± 0.03%), (R)-limonene (6.15 ± 0.04%), and linalyl acetate (5.02 ± 0.03%), which might be responsible for its observed anti-inflammation activity. It is concluded that CAVAO has great potential to be developed into a functional food for the treatment of inflammatory-associated diseases.

Influence of Environmental Factors on Essential Oil Variability in Origanum compactum Benth. Growing Wild in Morocco.

The present study aimed to evaluate the influence of environmental factors on essential oils (EOs) composition of Origanum compactum populations sampled all over the distribution area of the species in Morocco, and to determine the extent of the chemical profiles throughout the geographical distribution of the species. The chemical compositions were submitted to canonical correlation analysis and canonical discriminant analysis that indicated a significant relationship between oil components and some environmental factors. According to their chemical composition and edapho-climatic characteristics, two major groups of populations were differentiated. The first group was composed of samples growing in regions with humid climate, clayey, sandy, and alkaline soils. These samples showed high thymol, α-terpineol, linalool, and carvacryl methyl oxide content. The second group consisted of plants belonging to semi-arid climate, and growing at high altitudes and silty soils. These samples were characterized by high carvacrol, α-thujene, α-terpinene, and myrcene content. However, populations exposed to sub-humid climate, appeared less homogeneous and belong mainly either to the first or second group. A significant correlation between some edaphic factors (pH, K2 O content, soil texture) and the EOs yield of O. compactum plants was evidenced. In spite of the correlation obtained for the oil composition with edapho-climatic factors and the variance explained by the environmental data set, the observed EO diversity might be also genetically determined.

Glass bottle sampling solid phase microextraction gas chromatography mass spectrometry for breath analysis of drug metabolites.

Breath analysis is a non-invasive approach which may be applied to disease diagnosis and pharmacokinetic study. In the case of offline analysis, the exhaled gas needs to be collected and the sampling bag is often used as the storage vessel. However, the sampling bag usually releases some extra compounds, which may interfere with the result of the breath test. In this study, a novel breath sampling glass bottle was developed with a syringe needle sampling port for solid phase microextraction (SPME). Such a glass bottle scarcely liberates compounds and can be used to collect exhaled gas for ensuing analysis by gas chromatography-mass spectrometry (GC-MS). The glass bottle sampling SPME-GC-MS analysis was carried out to investigate the breath metabolites of myrtol, a multicompound drug normally used in the treatment of bronchitis and sinusitis. Four compounds, α-pinene, 2,3-dehydro-1,8-cineole, d-limonene and 1,8-cineole were found in the exhaled breath of all eight volunteers who had taken the myrtol. While for other ten subjects who had not used the myrtol, these compounds were undetectable. In the SPME-GC-MS analysis of the headspace of myrtol, three compounds were detected including α-pinene, d-limonene and 1,8-cineole. Comparing the results of breath and headspace analysis, it indicates that 2,3-dehydro-1,8-cineole in the breath is the metabolite of 1,8-cineole. It is the first time that this metabolite was identified in human breath. The study demonstrates that the glass bottle sampling SPME-GC-MS method is applicable to exhaled gas analysis including breath metabolites investigation of drugs like myrtol.

Converting citrus wastes into value-added products: Economic and environmently friendly approaches.

Citrus fruits, including oranges, grapefruits, lemons, limes, tangerines, and mandarins, are among the most widely cultivated fruits around the globe. Its production is increasing every year due to rising consumer demand. Citrus-processing industries generate huge amounts of wastes every year, and citrus peel waste alone accounts for almost 50% of the wet fruit mass. Citrus waste is of immense economic value as it contains an abundance of various flavonoids, carotenoids, dietary fiber, sugars, polyphenols, essential oils, and ascorbic acid, as well as considerable amounts of some trace elements. Citrus waste also contains high levels of sugars suitable for fermentation for bioethanol production. However, compounds such as D-limonene must be removed for efficient bioethanol production. The aim of the present article was to review the latest advances in various popular methods of extraction for obtaining value-added products from citrus waste/byproducts and their potential utility as a source of various functional compounds.

Indoor secondary organic aerosols formation from ozonolysis of monoterpene: An example of d-limonene with ammonia and potential impacts on pulmonary inflammations.

Monoterpene is one class of biogenic volatile organic compounds (BVOCs) which widely presents in household cleaning products and air fresheners. It plays reactive role in secondary organic aerosols (SOAs) formation with ozone (O3) in indoor environments. Such ozonolysis can be influenced by the presence of gaseous pollutants such as ammonia (NH3). This study focuses on investigations of ozone-initiated formation of indoor SOAs with d-limonene, one of the most abundant indoor monoterpenes, in a large environmental chamber. The maximum total particle number concentration from the ozonolysis in the presence of NH3 was 60% higher than that in the absence of NH3. Both of the nuclei coagulation and condensation involve in the SOAs growth. The potential risks of pulmonary injury for the exposure to the secondary particles formed were presented with the indexes of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) expression levels in bronchoalveolar lavage fluid (BALF) upon intratracheal instillation in mice lung for 6 and 12h. The results indicated that there was 22-39% stronger pulmonary inflammatory effect on the particles generated with NH3. This is a pilot study which demonstrates the toxicities of the indoor SOAs formed from the ozonolysis of a monoterpene.

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.

Seasonal variation of the essential oil from two Brazilian native Aldama La Llave (Asteraceae) species.

Aldama arenaria and A. robusta are morphologically similar aromatic species that have seasonal development. The yield and chemical composition of essential oils from aerial and underground vegetative organs of these species were compared to verify the production of volatile metabolites in flowering and dormant phases of development and to identify if there are unique compounds for either species. The major compound in the essential oils from A. arenaria leaves was palustrol (16.22%) and for aerial stems was limonene (15.3%), whereas limonene (11.16%) and α-pinene (19.64%) were the major compounds for leaves and aerial stems from A. robusta, respectively. The major compound for the underground organs was α-pinene, in both species and phenological stages. High amounts of diterpenes were found especially for A. arenaria essential oils. Each analyzed species presented unique compounds, which can provide a characteristic chemical profile for both species helping to solve their taxonomic problems. This study characterized for the first time the yield and essential oil composition of A. arenaria and A. robusta, which have medicinal potential, and some of the compounds in their essential oils are unique to each one and may be useful in helping the correct identification of them.

Bioactive chemical constituents of Curcuma longa L. rhizomes extract inhibit the growth of human hepatoma cell line (HepG2).

The present study was designed to identify the chemical constituents of the methanolic extract of Curcuma longa L. rhizomes and their inhibitory effect on a hepatoma cell line. The methanolic extract was subjected to GC-MS analysis to identify the volatile constituents and the other part of the same extract was subjected to liquid column chromatographic separation to isolate curcumin. The inhibition of cell growth in the hepatoma cell line and the cytopathological changes were studied. GC-MS analysis showed the presence of fifty compounds in the methanolic extract of C. longa. The major compounds were ar-turmerone (20.50 %), ß-sesquiphellandrene (5.20 %) and curcumenol (5.11 %). Curcumin was identified using IR, 1H and 13C NMR. The inhibition of cell growth by curcumin (IC50 = 41.69 ± 2.87 µg mL-1) was much more effective than that of methanolic extract (IC50 = 196.12 ± 5.25 µg mL-1). Degenerative and apoptotic changes were more evident in curcumin- treated hepatoma cells than in those treated with the methanol extract. Antitumor potential of the methanolic extract may be attributed to the presence of sesquiterpenes and phenolic constituents including curcumin (0.051 %, 511.39 µg g-1 dried methanol extract) in C. longa rhizomes.