pulegone and menthone

Topical drug delivery using nanoemulsions and nanogels is a promising approach to treating skin disorders such as melanoma.. In this study, the chemical composition of Mentha pulegium essential oil with five major compounds, including pulegone (68.11%), l-menthone (8.83%), limonene (2.90%), iso-pulegone (2.69%), and iso-menthone (1.48%) was first identified using GC-MS (Gas chromatography-Mass Spectrometry) analysis. Afterward, a nano-scaled emulsion containing the essential oil with a droplet size of 7.70 ± 1 nm was prepared. Nanogel containing the essential oil was then prepared by adding (2% w/v) carboxymethyl cellulose to the nano-scaled emulsion. Moreover, the successful loading of M. pulegium essential oil in the nano-scaled emulsion and nanogel was confirmed using ATR-FTIR (Attenuated total reflectance-Fourier Transform InfraRed) analysis. Then, human A375 melanoma cells were treated with different concentrations of samples, the MTT assay evaluated cell viability, and cell apoptosis was confirmed by flow cytometry. In addition, the expression of apoptotic and anti-apoptotic genes, including Bax and Bcl-2, was evaluated using the qPCR (quantitative Polymerase Chain Reaction) technique.. The results showed that cell viability was reduced by 90 and 45% after treatment with 300 μg/mL of the nanogel and nano-scaled emulsion. As confirmed by flow cytometry, this effect was mediated by apoptosis. Furthermore, gene expression analysis showed up-regulation of Bax and down-regulation of Bcl-2 genes. Therefore, the prepared nanogel, with high efficacy, could be considered a potent anticancer agent for supplementary medicine and in vivo research.

Topics: Apoptosis; bcl-2-Associated X Protein; Emulsions; Genes, Regulator; Humans; Melanoma; Mentha pulegium; Nanogels; Oils, Volatile

Our study evaluated the in vitro antioxidant properties, antibacterial and antifungal activities, anti-inflammatory properties, and chemical composition of the essential oils (EOs), total phenol, and total flavonoid of wild

Topics: Animals; Anti-Infective Agents; Antioxidants; Cyclohexane Monoterpenes; Flavonoids; Gas Chromatography-Mass Spectrometry; Humans; Male; Mentha pulegium; Menthol; Mice; Minerals; Oils, Volatile; Plant Extracts; Polyphenols

A novel automated method was developed for the quantitative determination of nine terpenoids that could contribute to the minty notes of red wine bouquet. The method couples headspace SPME-Arrow extraction with GC-MS/MS analysis. PDMS/DVB fiber was chosen for the extraction and an ionization energy of 30 eV permitted to optimize the analyte detection. The optimal sample preparation consists of a two-fold dilution of the wine sample with addition of 4 g of sodium chloride while the most suitable extraction conditions take place at 50 °C for 1 h. The method shows good linearity, intraday variations between 2 and 25%, interday variations between 7 and 23% and recoveries between 80 and 119%. The method exhibits the required low detection (between 3 and 60 ng/L) and quantification (between 6 ng/L and 200 ng/L) limits. These limits have permitted the quantification of the pool of minty terpenoids in fourteen red Bordeaux wines.

Topics: Cyclohexane Monoterpenes; Dimethylpolysiloxanes; Eucalyptol; Gas Chromatography-Mass Spectrometry; Lactones; Limonene; Mentha; Menthol; Odorants; Polyvinyls; Solid Phase Microextraction; Tandem Mass Spectrometry; Terpenes; Wine

The aerial parts of wild and cultivated Mentha mozaffarianii Jamzad were collected at full flowering stage from two provinces (Hormozgan and Fars) of Iran. The essential oils were extracted by a Clevenger approach and analysed using GC and GC-MS. The main components in wild plants were piperitenone (33.85%), piperitone (21.18%), linalool (6.89%), pulegone (5.93%), 1, 8.cineole (5.49%), piperitenone oxide (5.17%) and menthone (4.69%) and in cultivated plants, cis-piperitone epoxide (28.89%), linalool (15.36%), piperitone (11.57%), piperitenone oxide (10.14%), piperitenone (8.42%),1,8-cineole (3.60%) were the main constituents in essential oil. The in vitro antimicrobial activity of the essential oil of M. mozaffarianii was studied against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Candida albicans. The results of the bioassays showed that the oil exhibited high antimicrobial activity against all the tested pathogens.

Topics: Acyclic Monoterpenes; Anti-Infective Agents; Candida albicans; Cyclohexane Monoterpenes; Cyclohexanols; Eucalyptol; Gas Chromatography-Mass Spectrometry; Iran; Mentha; Menthol; Microbial Sensitivity Tests; Monoterpenes; Oils, Volatile; Plant Components, Aerial; Plants, Medicinal; Staphylococcus aureus

Schizonepeta tenuifolia Briquet, commonly known as Japanese catnip, is used for the treatment of colds, headaches, fevers, and skin rashes in traditional Asian medicine (China, Japan and Korea). The volatile oil and its constituents have various demonstrated biological activities, but there is currently limited information regarding the site of biosynthesis. Light microscopy and scanning electron microscopy indicated the presence of three distinct glandular trichome types which, based on their morphological features, are referred to as peltate, capitate and digitiform glandular trichomes. Laser scanning microscopy and 3D reconstruction demonstrated that terpenoid-producing peltate glandular trichomes contain a disk of twelve secretory cells. The oil of peltate glandular trichomes, collected by laser microdissection or using custom-made micropipettes, was demonstrated to contain (-)-pulegone, (+)-menthone and (+)-limonene as major constituents. Digitiform and capitate glandular trichomes did not contain appreciable levels of terpenoid volatiles. The yield of distilled oil from spikes was significantly (44%) higher than that from leaves, while the composition of oils was very similar. Oils collected directly from leaf peltate glandular trichomes over the course of a growing season contained primarily (-)-pulegone (>80% at 32 days after germination) in young plants, while (+)-menthone began to accumulate later (>75% at 80 days after germination), at the expense of (-)-pulegone (the levels of (+)-limonene remained fairly stable at 3-5%). The current study establishes the morphological and chemical characteristics of glandular trichome types of S. tenuifolia, and also provides the basis for unraveling the biosynthesis of essential oil in this popular medicinal plant.

Topics: Chromatography, Gas; Cyclohexane Monoterpenes; Cyclohexenes; Japan; Limonene; Medicine, Traditional; Menthol; Monoterpenes; Nepeta; Oils, Volatile; Plant Leaves; Plants, Medicinal; Stereoisomerism; Terpenes; Trichomes

Human noroviruses are the causative agents of non-bacterial gastroenteritis worldwide. The rapid onset and resolution of disease symptoms suggest that innate immune responses are critical for controlling norovirus infection; however, no effective antivirals are yet available. The present study was conducted to examine the antiviral activities of Schizonepeta tenuifolia Briquet extract (STE) against noroviruses. Treatment of human norovirus replicon-bearing HG23 cells with STE at 5 and 10 mg/ml concentrations resulted in the reduction in the viral RNA levels by 77.2% and 85.9%, respectively. STE had no cytotoxic effects on HG23 cells. Treatment of RAW 264.7 cells infected with murine norovirus 1 (MNV-1), a surrogate virus of human noroviruses, with STE at 10 and 20 µg/ml concentrations resulted in the reduction of viral replication by 58.5% and 84.9%, respectively. STE treatment induced the expression of mRNAs for type I and type II interferons in HG23 cells and upregulated the transcription of interferon-β in infected RAW 264.7 cells via increased phosphorylation of interferon regulatory factor 3, a critical transcription regulator for type I interferon production. These results suggest that STE inhibits norovirus replication through the induction of antiviral interferon production during virus replication and may serve as a candidate antiviral substance for treatment against noroviruses.

Topics: Animals; Antiviral Agents; Caliciviridae Infections; Cell Line; Cyclohexane Monoterpenes; Humans; Immunity, Innate; Interferon Regulatory Factor-3; Interferon Type I; Interferon-gamma; Interferons; Lamiaceae; Menthol; Mice; Monoterpenes; Norovirus; Norwalk virus; Phytochemicals; Plant Extracts; Polycyclic Sesquiterpenes; RAW 264.7 Cells; RNA, Viral; Sesquiterpenes; Virus Replication

A method was established for separation, purification, enrichment and enumeration of peltate glandular trichomes from Schizonepetae Spica which combined with qualitative and quantitative analysis of major constituents of its inclusions.. Frozen brush method and sedimentation of water concentration connected to technology of materials affinity were applied to separate and concentrate peltate glandular trichomes respectively, and extracts of inclusions was qualified by GCMS. Microscope enumeration was considered used to quantitatively analyze three monoterperes in single peltate glandular trichome and the calyx of Schizonepetae Spica.. 29 chemical compounds were identified from peltate glandular trichomes’ inclusions of Schizonepetae Spica,constituting 97. 45% of the total peak area. 35 characteristic common peaks exceeding 97% of the total peak area was both present in peltate glandular trichomes’ inclusions and essential oils of Schizonepetae Spica. The difference contents of d-limonene, dl-menthone, pulegone, E-carveol acetate, et al between peltate glandular trichomes’ inclusions and essential oils of Schizonepetae Spica were large. The content of d-limonene,dl-menthone, pulgone in a single peltate glandular trichome was 27. 660,24. 334 and 396. 390 ng respectively through the method of microscope enumeration.. The research provides a method of separation, purification enrichment and enumeration of peltate glandular trichomes. The chemical constituent of essential oil which is extracted by water vapor is different from peltate glandular trichomes’ inclusions and could not reflect the ratio and content, while prediction model which based on microscope enumeration is precision.

Topics: Cyclohexane Monoterpenes; Cyclohexenes; Flowers; Limonene; Menthol; Microscopy; Monoterpenes; Oils, Volatile; Plant Leaves; Terpenes; Trichomes

Medicinal plants are known as important sources of secondary metabolites. Because of the economic value of pennyroyal [Mentha pulegium L. (Lamiaceae)] in food industries, propagation of this valuable plant has special importance. Plant cell suspension culture can increase some produced components. The aim of this research was performing cell culture for induction of some secondary metabolites of M. pulegium and compares it with native one. The MS medium was used for suspension culture. To investigate quantitative materials, 4 levels of yeast extract elicitor (20, 40, 60 and 80 mg/L) and salicylic acid in 4 levels (2, 4, 6 and 8 mg/L) were used. Obtained extracts were analyzed by GC-MS. Statistical analysis showed that the amount of limonene, menthone, menthol and α-pinene were more than mentioned compounds in natural plant as control. The maximum amount of this metabolites were obtained as limonene (in 60 mg/l yeast extract), menthone (in 40 mg/l yeast extract and 2 mg/l salicylic acid), menthol (in 6 mg/l salicylic acid) and α-pinene (in 4 mg/l salicylic acid) in the M. pulegium cell culture. The Pulegone was fond more in natural plants than cell culture mass. The most important secondary metabolites were increased by cell culture containing of salicylic acid and yeast extract elicitors in M. pulegume.

Topics: Bicyclic Monoterpenes; Biotechnology; Cell Culture Techniques; Cyclohexane Monoterpenes; Cyclohexenes; Limonene; Mentha pulegium; Menthol; Monoterpenes; Salicylic Acid; Terpenes; Yeasts

In vitro conditions and benzyladenine influenced both content and composition of micropropagated Micromeria pulegium essential oils, with pulegone and menthone being the main essential oil components. The content and chemical composition of Micromeria pulegium (Rochel) Benth. essential oils were studied in native plant material at vegetative stage and in micropropagated plants, obtained from nodal segments cultured on solid MS medium supplemented with N(6)-benzyladenine (BA) or kinetin at different concentrations, alone or in combination with indole-3-acetic acid. Shoot proliferation was achieved in all treatments, but the highest biomass production was obtained after treatment with 10 μM BA. Phytochemical analysis identified up to 21 compounds in the essential oils of wild-growing and in vitro cultivated plants, both showing very high percentages of total monoterpenoids dominated by oxygenated monoterpenes of the menthane type. Pulegone and menthone were the main essential oil components detected in both wild-growing plants (60.07 and 26.85 %, respectively) and micropropagated plants grown on either plant growth regulator-free medium (44.57 and 29.14 %, respectively) or BA-supplemented medium (50.77 and 14.45 %, respectively). The percentage of total sesquiterpenoids increased in vitro, particularly owing to sesquiterpene hydrocarbons that were not found in wild-growing plants. Differences in both content and the composition of the essential oils obtained from different samples indicated that in vitro culture conditions and plant growth regulators significantly influence the essential oils properties. In addition, the morphology and structure of M. pulegium glandular trichomes in relation to the secretory process were characterized for the first time using SEM and light microscopy, and their secretion was histochemically analyzed.

Topics: Adenine; Culture Techniques; Cyclohexane Monoterpenes; Kinetin; Lamiaceae; Menthol; Microscopy, Electron, Scanning; Monoterpenes; Oils, Volatile; Trichomes

Minthostachys verticillata (Lamiaceae), popularly known as peperina is largely used in popular medicine for its digestive, carminative, antispasmodic and antirheumatic properties. There are no reports of repeated exposure toxicity to guarantee their safety. The present study investigated the chemical composition, analyzed by GC-FID, and the 90-day toxicity and genotoxicity effect of M. verticillata essential oil (Mv-EO), using Wistar rats as test animals. The rats were divided into four groups (5 rats/sex/group) and Mv-EO was administered on diet at doses of 0, 1, 4 and 7 g/kg feed. The main components of Mv-EO were pulegone (64.65%) and menthone (23.92%). There was no mortality, adverse effects on general conditions or changes in body weight, food consumption and feed conversion efficiency throughout the study in male and female rats. Subchronic administration of Mv-EO did not alter the weights, morphological and histopathological analyses of liver, kidney and intestine. Genotoxicity was tested by micronucleus and comet assays. Mv-EO up to a concentration of 7 g/kg feed for 90 days did not exert a cyto-genotoxic effect on the bone marrow and cells blood of Wistar rats. These results suggest that Mv-EO appears to be safe and could be devoid of any toxic risk.

Topics: Animals; Blood Cells; Bone Marrow; Comet Assay; Cyclohexane Monoterpenes; Female; Intestines; Kidney; Lamiaceae; Liver; Male; Menthol; Micronucleus Tests; Monoterpenes; Oils, Volatile; Rats, Wistar; Toxicity Tests, Subchronic; Weight Gain

Danshu capsule (DSC) is a medicinal compound in traditional Chinese medicine (TCM). It is commonly used for the treatment of acute & chronic cholecystitis as well as choleithiasis. To study its choleretic effect, healthy rats were randomly divided into DSC high (DSCH, 900mg/kg), medium (DSCM, 450mg/kg), and low (DSCL, 225mg/kg) group, Xiaoyan Lidan tablet (XYLDT, 750mg/kg), and saline group. The bile was collected for 1h after 20-minute stabilization as the base level, and at 1h, 2h, 3h, and 4h after drug administration, respectively. Bile volume, total cholesterol, and total bile acid were measured at each time point. The results revealed that DSC significantly stimulated bile secretion, decreased total cholesterol level and increased total bile acid level. Therefore, it had choleretic effects. To identify the active components contributing to its choleretic effects, five major constituents which are menthol (39.33mg/kg), menthone (18.02mg/kg), isomenthone (8.18mg/kg), pluegone (3.31mg/kg), and limonene (4.39mg/kg) were tested on our rat model. The results showed that menthol and limonene could promote bile secretion when compared to DSC treatment (p > 0.05); Menthol, menthol and limonene could significantly decrease total cholesterol level (p<0.05 or p<0.01) as well as increase total bile acid level (p<0.05 or p<0.01); Isomenthone, as a isomer of menthone, existed slightly choleretic effects; Pluegone had no obvious role in bile acid efflux. These findings indicated that the choleretic effects of DSC may be attributed mainly to its three major constituents: menthol, menthone and limonene.

Topics: Animals; Bile; Bile Acids and Salts; Cholagogues and Choleretics; Cholecystitis; Cholelithiasis; Cholesterol; Complex Mixtures; Cyclohexane Monoterpenes; Cyclohexenes; Humans; Limonene; Male; Medicine, Chinese Traditional; Menthol; Monoterpenes; Rats; Rats, Sprague-Dawley; Terpenes

The chemical composition of essential oil obtained by hydrodistillation of the fresh aerial parts of Mentha requienii Bentham (Lamiaceae) collected on the Gennargentu Mountains (Sardinia, Italy) has been investigated by gas chromatography and gas chromatography-mass spectrometry. The main constituents that resulted were pulegone (78%), menthone (0.5%), isomenthone (18%), isopulegone (1.3%) and limonene (1.76%). In vitro antifungal activity is evaluated in order to identify new means that could be helpful in the prevention of contamination in indoor environments.

Topics: Antifungal Agents; Chromatography, Gas; Cyclohexane Monoterpenes; Cyclohexenes; Gas Chromatography-Mass Spectrometry; Italy; Limonene; Mentha; Menthol; Microbial Sensitivity Tests; Mitosporic Fungi; Monoterpenes; Oils, Volatile; Plant Components, Aerial; Plant Extracts; Terpenes

The hydro-distilled essential oils obtained from aerial parts of the wild (w) and cultivated (c) mint timija (Mentha suaveolens subsp. timija), an endemic medicinal species of Morocco, have been analyzed by GC-MS and screened for antimicrobial activity. In total, 35 compounds representing more than 98% of the oils were identified. Menthone (39.4(w)-10.8(c)%), pulegone (62.3(c)-34.3(w)%) and isomenthone (9.3(c)-7.8(w)%) were found as the main components for the two oils. The volatiles of the wild and cultivated material differed significantly in both the percentage of the main components and antimicrobial effect. Pulegone was more dominant in cultivated mint timija (62.3%) than in wild one (34.3%), while menthone was more abundant in the wild material (39.4%). In the antimicrobial assays, both oils displayed good to excellent activity against all microorganisms tested with the oil of the cultivated form being more active.

Topics: Anti-Infective Agents; Cyclohexane Monoterpenes; Drug Evaluation, Preclinical; Endangered Species; Gas Chromatography-Mass Spectrometry; Mentha; Menthol; Microbial Sensitivity Tests; Monoterpenes; Morocco; Oils, Volatile; Plant Components, Aerial

Transient receptor potential (TRP) vanilloid-1 (TRPV1) and melastatin-8 (TRPM8) channels play a role in transmitting sensory information in primary-afferent neurons. TRPV1 agonists at high concentrations inhibit action potential conduction in the neurons and thus have a local anesthetic effect. The purpose of the present study was to know whether TRPM8 agonist menthol at high concentrations has a similar action and if so whether there is a structure-activity relationship among menthol-related chemicals.. Compound action potentials (CAPs) were recorded from the frog sciatic nerve by using the air-gap method.. (-)-Menthol and (+)-menthol concentration-dependently reduced CAP peak amplitude with the IC(50) values of 1.1 and 0.93 mM, respectively. This (-)-menthol activity was resistant to non-selective TRP antagonist ruthenium red; TRPM8 agonist icilin did not affect CAPs, indicating no involvements of TRPM8 channels. p-Menthane, (+)-limonene and menthyl chloride at 7-10 mM minimally affected CAPs. On the other hand, (-)-menthone, (+)-menthone, (-)-carvone, (+)-carvone and (-)-carveol (in each of which chemicals OH or O group was added to p-menthane and limonene) and (+)-pulegone inhibited CAPs with extents similar to that of menthol. 1,8-Cineole and 1,4-cineole were less effective while thymol and carvacrol were more effective than menthol in inhibiting CAPs.. Menthol-related chemicals inhibited CAPs and were thus suggested to exhibit local anesthetic effects comparable to those of lidocaine and cocaine as reported previously for frog CAPs. This result may provide information to develop local anesthetics on the basis of the chemical structure of menthol.

Topics: Action Potentials; Animals; Antipruritics; Cyclohexane Monoterpenes; Cyclohexanes; Cyclohexanols; Cyclohexenes; Cymenes; Dose-Response Relationship, Drug; Eucalyptol; Female; Limonene; Male; Menthol; Monoterpenes; Ranidae; Sciatic Nerve; Structure-Activity Relationship; Terpenes; Thymol; TRPV Cation Channels

Intense applications of synthetic insecticides for the control of adult Musca domestica have led to the insects developing resistance to most of them. In consequence, there is interest in new active ingredients as alternatives to conventional insecticides. Essential oils (EO) are potential tools for controlling M. domestica because of their effectiveness and their minimal environmental effects. In a fumigant assay, M. domestica adults treated with Minthostachys verticillata EO [LC(50)=0.5 mg/dm(3); majority components by SPME-GC: (4R)(+)-pulegone (67.5%), menthone (22.3%) and (4R)(+)-limonene (3.8%)], died within 15 min or less. The terpenes absorbed by the flies and their metabolites, analyzed using SPME fiber, were (4R)(+)-limonene (LC(50)=6.2 mg/dm(3)), menthone (LC(50)=1.9 mg/dm(3)), (4R)(+)-pulegone (LC(50)=1.7 mg/dm(3)) and a new component, menthofuran (LC(50)=0.3 mg/dm(3)), in a relative proportion of 12.4, 6.5, 35.9 and 44.2% respectively. Menthofuran was formed by oxidation of either (4R)(+)-pulegone or menthone mediated by cytochrome P450, as demonstrated by a fumigation assay on flies previously treated with piperonyl butoxide, a P450 inhibitor, which showed a decrease in toxicity of the EO, (4R)(+)-pulegone and of menthone, supporting the participation of the P450 oxidizing system in the formation of menthofuran. The enzymatic reaction of isolated fly microsomes with the EO or the (4R)(+)-pulegone produced menthofuran in both cases. Contrary to expectations, the insect detoxification system contributed to enhance the toxicity of the M. verticillata EO. Consequently, resistant strains overexpressing P450 genes will be more susceptible to either M. verticillata EO or (4R)(+)-pulegone and menthone.

Topics: Animals; Biological Assay; Cell Survival; Cyclohexane Monoterpenes; Cytochrome P-450 Enzyme System; Drug Synergism; Houseflies; Insecticides; Lamiaceae; Lethal Dose 50; Menthol; Monoterpenes; Oils, Volatile; Plant Extracts; Plant Leaves; Plant Oils; Terpenes

Populations of Mentha longifolia, an endangered species in Israel, were tested for essential oil composition and conservational ability. In 2002-2003, 25 wild populations country-wide were tested, indicating population divergence into two chemotypes. Chemotype A was characterized by high levels of menthone and pulegone, and chemotype B by high levels of piperitenone oxide and piperitone oxide. Chemotype A was more abundant (22 of 25 populations) than chemotype B (11 of 25 populations). However, a chemotype/population interaction was not recorded (P > 0.05). In spring 2003, seven of the 25 wild populations were resampled, propagated, and cultivated at the Newe Ya'ar campus. Then, in 2004, the propagated plants were tested for essential oil composition. The propagated plants maintained the essential oil composition as well as the chemotype-frequency distribution of the original wild population from which they were obtained. Since a chemotype/population interaction was not recorded, and the cultivated plants displayed the wild population essential oil composition, it can be concluded that i) the chemotype diversity is genetically based, and ii) the M. longifolia populations sampled can be horticulturally conserved.

Topics: Cyclohexane Monoterpenes; Israel; Mentha; Menthol; Monoterpenes; Oils, Volatile

The essential oils (EOs) and static headspaces (HSs) of in vitro plantlets and callus of Mentha x piperita were characterized by GC-MS analysis. Leaves were used as explants to induce in vitro plant material. The EO yields of the in vitro biomass were much lower (0.1% v/w) than those of the parent plants (2% v/w). Many typical mint volatiles were emitted by the in vitro production, but the callus and in vitro plantelet EOs were characterized by the lack of both pulegone and menthofuran. This was an important difference between in vitro and in vivo plant material as huge amounts of pulegone and menthofuran may jeopardise the safety of mint essential oil. Regarding the other characteristic volatiles, menthone was present in reduced amounts (2%) in the in vitro plantlets and was not detected in the callus, even if it represented the main constituent of the stem and leaf EOs obtained from the cultivated mint (26% leaves; 33% stems). The M. piperita callus was characterized by menthol (9%) and menthone (2%), while the in vitro plantlet EO showed lower amounts of both these compounds in favour of piperitenone oxide (45%). Therefore, the established callus and in vitro plantlets showed peculiar aromatic profiles characterized by the lack of pulegone and menthofuran which have to be monitored in the mint oil for their toxicity.

Topics: Cell Culture Techniques; Cyclohexane Monoterpenes; Gas Chromatography-Mass Spectrometry; Mentha piperita; Menthol; Monoterpenes; Oils, Volatile; Plant Leaves; Plant Oils; Plant Stems; Volatile Organic Compounds

The essential oil derived from the flowering aerial parts of Schizonpeta multifida was investigated for insecticidal activity against two grain storage insects, maize weevil (Sitophilus zeamais) and red flour beetle (Tribolium castaneum).. The main constituents of the essential oil were menthone (40.34%) and pulegone (26.87%), followed by D-limonene (5.81%) and isomenthone (5.14%). Bioactivity-directed chromatographic separation on repeated silica gel columns led to the isolation of menthone and pulegone. Pulegone showed pronounced contact toxicity against S. zeamais and T. castaneum adults (7 day LD(50)= 4.46 and 1.55 µg per adult respectively) and was more toxic than menthone (7 day LD(50)=33.47 and 2.67 µg per adult respectively), while the crude essential oil had 7 day LD(50) values of 30.17 and 2.75 µg per adult respectively. Pulegone also displayed strong fumigant toxicity against S. zeamais and T. castaneum adults (7 day LC(50)=3.47 and 11.56 mg cm(-3) respectively) and was again more toxic than menthone (7 day L(50) =10.32 and 31.25 mg cm(-3) respectively), while the crude essential oil had 7 day LC(50) values of 8.33 and 26.41 mg cm(-3) respectively.. The essential oil of S. multifida and its two main components may have potential to be developed as new natural fumigants/insecticides for the control of stored product insects.

Topics: Animals; Cyclohexane Monoterpenes; Fumigation; Insecticides; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Plant Components, Aerial; Tribolium; Weevils

In a previous work, we have demonstrated that Minthostachys verticillata essential oil has immunomodulatory effects in vitro on cells from allergic patients. Here we characterized main components of M. verticillata essential oil and also tested if these compounds modulate In vitro and in vivo the immediate-type allergic reaction. Gas chromatography was used to identify main components of the essential oil. Pulegone (63.4 %), menthone (15.9 %), and limonene (2.1 %) were found as main classes. IL-13 levels were evaluated from lymphocytes cultures stimulated with allergen alone or combined with monoterpenes. All compounds stimulated cell proliferation but, interestingly, promoted a reduction of IL-13 values, limonene and the mixture of the three compounds being the most active. β-Hexosaminidase release was determined from basophils to which essential oil or monoterpenes were added. We observed that, whichever combination of monoterpenes was used, β-hexosaminidase release was diminished in all cases. Obtained values were even lower than those of antiallergic drug desloratadine. Essential oil and limonene inhibited mast cell activation and degranulation in the skin when testing passive cutaneous anaphylaxis, limonene being the most active. In conclusion, limonene was the compound that showed the most potent immunomodulatory activity. This fact suggests that it constitutes a promising natural alternative for a novel treatment of allergic diseases.

Topics: Adolescent; Adult; Animals; Anti-Allergic Agents; Basophils; beta-N-Acetylhexosaminidases; Cell Proliferation; Child; Child, Preschool; Cyclohexane Monoterpenes; Cyclohexenes; Dose-Response Relationship, Drug; Female; Humans; Hypersensitivity, Immediate; Infant; Interleukin-13; Lamiaceae; Leukocytes, Mononuclear; Limonene; Male; Mast Cells; Menthol; Mice; Mice, Inbred BALB C; Monoterpenes; Oils, Volatile; Terpenes; Young Adult

The chemical composition of the essential oils of Ziziphora clinopodioides Lam. from the aerial flowering parts, collected during two different years, were obtained by hydrodistillation and analyzed by gas chromatography - mass spectrometry. Forty-five components representing 100% and 94.7% of the total oil were identified. The main constituents of the essential oils were pulegone (72.8 and 35.0%), neomenthol (6.5 and 23.1%), menthone (6.2 and 13.3%), p-menth-3-en-8-ol (1.7 and 3.5%), piperitenone (2.6 and 1.1%) and piperitone (0.7 and 1.2%). A cluster analysis was carried out on the essential oil compositions of Z. clinopodioides.

Topics: Cyclohexane Monoterpenes; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Plants, Medicinal; Tajikistan

Volatile organic compounds (VOCs), characterized by low molecular weight and high vapor pressure, are produced by all organisms as part of normal metabolism, and play important roles in communication within and between organisms. We examined the effects of VOCs released by three species of plant growth-promoting rhizobacteria (Pseudomonas fluorescens, Bacillus subtilis, Azospirillum brasilense) on growth parameters and composition of essential oils (EO) in the aromatic plant Mentha piperita (peppermint). The bacteria and plants were grown on the same Petri dish, but were separated by a physical barrier such that the plants were exposed only to VOCs but not to solutes from the bacteria. Growth parameters of plants exposed to VOCs of P. fluorescens or B. subtilis were significantly higher than those of controls or A. brasilense-treated plants. Production of EOs (monoterpenes) was increased 2-fold in P. fluorescens-treated plants. Two major EOs, (+)pulegone and (-)menthone, showed increased biosynthesis in P. fluorescens-treated plants. Menthol in A. brasilense-treated plants was the only major EO that showed a significant decrease. These findings suggest that VOCs of rhizobacteria, besides inducing biosynthesis of secondary metabolites, affect pathway flux or specific steps of monoterpene metabolism. Bacterial VOCs are a rich source for new natural compounds that may increase crop productivity and EO yield of this economically important plant species.

Topics: Azospirillum brasilense; Bacillus subtilis; Biosynthetic Pathways; Culture Media; Culture Techniques; Cyclohexane Monoterpenes; Mentha piperita; Menthol; Monoterpenes; Oils, Volatile; Pseudomonas fluorescens; Volatile Organic Compounds

A GC-MS-Selected Ion Monitoring (SIM) detection method was developed for simultaneous determination of four monoterpenes: (-)-menthone, (+)-pulegone, (-)-limonene and (+)-menthofuran as the main bio-active and toxic constituents, and four other main compounds in the volatile oils of Schizonepeta tenuifolia (ST) leaves and spikes at different harvesting times. The results showed that the method was simple, sensitive and reproducible, and that harvesting time was a possible key factor in influencing the quality of ST leaves, but not its spikes. The research might be helpful for determining the harvesting time of ST samples and establishing a validated method for the quality control of ST volatile oil and other relative products.

Topics: Cyclohexane Monoterpenes; Gas Chromatography-Mass Spectrometry; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Plant Components, Aerial; Plant Extracts; Plant Leaves; Time Factors

The essential-oil profile of a Calamintha species, wild-growing in the urban settings of the city of Niš (South Serbia) and botanically tentatively identified as C. vardarensis (endemic species native to FYR Macedonia and East Serbia), has been statistically (multivariate statistical analyses) compared with those of other Calamintha species, including two previously investigated C. vardarensis populations, as a means of corroboration of the surprising occurence of this Calamintha population outside of its natural distributional range. Agglomerative hierarchical clustering reveals a close link of C. vardarensis from Niš (with neo-menthol (40.0%), menthone (21.8%), and pulegone (27.2%) as its major oil contributors) and C. vardarensis from FYR Macedonia.

Topics: Cyclohexane Monoterpenes; Gas Chromatography-Mass Spectrometry; Introduced Species; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Principal Component Analysis

This study addresses the adaptive dynamics of plant toxins that ward off herbivores, an aspect of chemical warfare in ecosystems. Thyme basil (Acinos suaveolens) growing in an area regularly grazed by mammals contains double the concentration of monoterpenoids (15.61 vs. 8.18 mg/g dry matter, P = 0.001) when contrasted with plants growing on inaccessible rock pillars and not exposed to mammalian herbivores. Thyme basil produces two monoterpenoids-menthone and R-(+)-pulegone-that show contrasting toxicity. The more toxic R-(+)-pulegone shows an increase in relative abundance (66.3% vs. 47.5%, P < 0.001) in plants exposed to browsing The results demonstrate how plant chemistry responds to both abiotic and biotic environmental conditions.

Topics: Animals; Cyclohexane Monoterpenes; Ecosystem; Environment; Mammals; Menthol; Monoterpenes; Plant Leaves; Thymus Plant

Pulegone is a major constituent of pennyroyal oil and a minor component of peppermint oil. Pulegone is biotransformed to menthofuran and menthones (diastereomeric menthone and isomenthone) in pennyroyal and peppermint as well as in rodents. Pulegone and menthofuran are hepatotoxic to rodents, and menthones are less toxic. The metabolism and disposition of pulegone and menthofuran were previously studied in rodents, and higher concentrations of pulegone- and menthofuran-derived radioactivity were observed in male than female rat kidney. One explanation is the association of pulegone and metabolites with a male rat-specific protein, alpha2u-globulin. To test this hypothesis, male and female rats were dosed orally with 14C-labeled pulegone (80 mg/kg, 120 microCi/kg) or menthofuran (60 mg/kg, 120 microCi/kg) or menthones (80 mg/kg, 120 microCi/kg) in corn oil, and the kidney cytosol was prepared 24 h after dosing. An equilibrium dialysis experiment showed that in all three studies the radioactivity was associated with kidney cytosol proteins of male but not female rats. The chemicals present in the male rat kidney cytosol after dialysis were extracted with dichloromethane and characterized by high-performance liquid chromatography (HPLC) and gas chromatography/mass spectrometry (GC-MS). All parent compounds were detected, and the metabolites characterized included piperitone from pulegone or menthones treatment, menthones and possibly 8-hydroxymenthones from pulegone treatment, and mintlactones (diastereomeric mintlactone and isomintlactone) and 7a-hydroxymintlactone from menthofuran treatment. Analysis of the male rat kidney cytosol by a gel filtration column demonstrated that the retention was due to reversible binding of these chemicals with the male rat-specific protein alpha2u-globulin. However, binding of pulegone and/or metabolites to alpha2u-globulin did not produce accumulation of this protein in the kidney.

Topics: Alpha-Globulins; Animals; Carbon Radioisotopes; Cyclohexane Monoterpenes; Female; Kidney Glomerulus; Male; Menthol; Metabolic Networks and Pathways; Monoterpenes; Rats; Sex Factors

Plants display a diverse array of inducible changes in secondary metabolites following insect herbivory. Herbivores differ in their feeding behavior, physiology, and mode of attachment to the leaf surface, and such variations might be reflected in the induced responses of damaged plants. Induced changes were analyzed for Minthostachys mollis, a Lamiaceae with medicinal and aromatic uses, and four species of folivore insects with different feeding habits (chewing, scraping, sap-sucking, and puncturing). In M. mollis leaves experimentally exposed to the insects, levels of the two dominant monoterpenes pulegone and menthone were assessed 24 and 48 h after wounding. Menthone content generally decreased in the essential oil of damaged leaves, whereas pulegone concentration increased in all treatments. These changes occurred also in the adjacent undamaged leaves, suggesting a systemic response. The relatively uniform response to different kinds of damage could be attributable to the presence of such a strongly active compound as pulegone in the essential oil of M. mollis. The effects of wounding on essential oil concentration may be significant from a commercial point of view.

Topics: Animals; Cyclohexane Monoterpenes; Feeding Behavior; Insecta; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Plant Leaves

Plant tissues may show chemical changes following damage. This possibility was analyzed for Minthostachys mollis, a Lamiaceae native to Central Argentina with medicinal and aromatic uses in the region. Effects of mechanical damage on its two dominant monoterpenes, pulegone and menthone, were analyzed by perforating M. mollis leaves and then assessing essential oil composition at 24, 48, and 120 hr; emission of volatiles was also measured 24 and 48 hr after wounding. Mechanical damage resulted in an increase of pulegone and menthone concentration in M. mollis essential oil during the first 24 hr. These changes did not occur in the adjacent undamaged leaves, suggesting a lack of systemic response. Postwounding changes in the volatiles released from M. mollis damaged leaves were also detected, most noticeably showing an increase in the emission of pulegone. Inducible chemical changes in aromatic plants might be common and widespread, affecting the specific compounds on which commercial exploitation is based.

Topics: Animals; Argentina; Cyclohexane Monoterpenes; Insecta; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Plant Leaves; Plant Oils

Essential oil was extracted from Schizonepeta tenuifolia Briq. by supercritical fluid extraction (SFE) and steam distillation (SD). The components extracted were determined by gas chromatography with area normalization method and identified by gas chromatography-mass spectrometry (GC-MS). The optimal chromatographic conditions were: capillary column, SE-54 (30 m x 0.25 mm i.d., 0.25 microm); column temperature, 50 degrees C (3 min) --> (5 degrees c/min) 180 degrees C (2 min --> (10 degrees C/ min) 260 degrees C 50 min); split injection, split ratio 1: 50; injector temperature, 280 degrees C. Fifty-four components were identified for the essential oils extracted by SFE, and its main components were found to be pulegone, menthone, linoleic acid chloride etc. Thirty-nine components were identified for the essential oil obtained by SD, and its main components were found to be pulegone, menthone, limonene etc. The SFE method is better than the SD method in reliability stability and reproducibility, and is thus well suitable for similar applications involving for extraction of other traditional Chinese herbal medicines.

Topics: Chromatography, Supercritical Fluid; Cyclohexane Monoterpenes; Distillation; Drugs, Chinese Herbal; Gas Chromatography-Mass Spectrometry; Lamiaceae; Linoleic Acid; Menthol; Monoterpenes; Oils, Volatile; Reproducibility of Results

The Fourier transform Raman (FT-Raman) spectra of pure terpenes and essential oils obtained by hydrodistillation of some Lamiaceae species, are presented. This study shows that principal components of an essential oil can be recognized by FT-Raman. Components predicted by FT-Raman spectrum of an essential oil correlate well with those found as major constituents by GC-MS. In this way the basic chemical character of an essential oil can be recognized. The results demonstrate that certain Raman intensities can be correlated to specific terpenes and therefore FT-Raman can discriminate between the essential oils of which main components belong to different classes of compounds.

Topics: Acyclic Monoterpenes; Camphor; Cyclohexane Monoterpenes; Cyclohexanols; Cyclohexanones; Eucalyptol; Fourier Analysis; Gas Chromatography-Mass Spectrometry; Lamiaceae; Lavandula; Menthol; Monoterpenes; Oils, Volatile; Plant Oils; Salvia officinalis; Spectrum Analysis, Raman; Terpenes

Plant tissues may show chemical changes following herbivory. In aromatic plants such changes could affect the specific compounds on which commercial exploitation is based. This possibility was analyzed for Mintosthachys mollis, a member of the Lamiaceae native to Central Argentina with medicinal and aromatic uses in the region, and two types of insect herbivores: a leaf miner and a gall insect. Analysis of the essential oils of mined/undamaged leaves, as well as leaves from stems with and without galls, revealed changes in concentrations of the two main monoterpenes. A decrease in pulegone concentration was associated with both types of insect damage, whereas menthone increased significantly only in mined leaves. Inducible chemical changes in aromatic and medicinal plants may be common and widespread; their economic implications deserve investigation.

Topics: Animals; Chromatography, Gas; Cyclohexane Monoterpenes; Diptera; Lamiaceae; Menthol; Monoterpenes; Oils, Volatile; Plant Leaves; Plant Oils; Terpenes

Aqueous solutions of [2H]-labeled pulegone enantiomers were fed to Mentha spicata ssp. spicata L. and Mentha spicata ssp. crispata L. shoot tip and first leaf pair. After solid phase microextraction the essential oil was analysed with enantioselective multidimensional gas chromatography/mass spectrometry. Both Mentha spicata species were able to convert labelled (1R)- and (1S)-pulegone at the same rate into the corresponding menthone and isomenthone, indicating an unspecific process. The reduction of both pulegone enantiomers preferably led to the cis-stereoisomers. The observed stereoselectivity is completely different from those of pulegone reduction by Mentha x piperita L.

Topics: Cyclohexane Monoterpenes; Deuterium; Gas Chromatography-Mass Spectrometry; Isotope Labeling; Lamiaceae; Menthol; Monoterpenes; Plant Leaves; Plant Shoots; Stereoisomerism; Terpenes

Mentha x piperita shoot tips and first leaf pairs were fed with aqueous solutions of different deuterium-labeled pulegone and various enantiomeric distributions. The essential oil was extracted by solid-phase microextraction and analyzed using enantioselective multidimensional gas chromatography/mass spectrometry. The genuine p-menthan-3-ones (-)-menthone and (+)-isomenthone as well as their labeled analogues were analyzed simultaneously. Both enantiomers of labeled pulegone were converted into the corresponding labeled p-menthan-3-ones by Mentha x piperita, indicating an unspecific reduction process. The generation of 4S- and 4R-configured p-menthan-3-ones differed in their stereoselectivities. Labeled (S)-pulegone was reduced by Mentha x piperita more rapidly rather than (R)-pulegone. From a comparison of labeled pulegone enantiomers the bioconversion preferrably led to 4S-configured diastereomers.

Topics: Biotransformation; Cyclohexane Monoterpenes; Isomerism; Lamiaceae; Magnoliopsida; Menthol; Monoterpenes; Terpenes

Topics: Charcoal; Cyclohexane Monoterpenes; Drug Stability; Hot Temperature; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Menthol; Monoterpenes; Oils, Volatile; Plants, Medicinal; Terpenes

Topics: Animals; Biotransformation; Cyclohexane Monoterpenes; Flavoring Agents; Menthol; Microsomes, Liver; Monoterpenes; Mutagenicity Tests; Mutation; Rats; Salmonella typhimurium; Terpenes