Rapid Classification and Recognition Method of the Species and Chemotypes of Essential Oils by ATR-FTIR Spectroscopy Coupled with Chemometrics.

In the present work, the applicability of attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, coupled with chemometric tools in recognizing essential oils (EOs) for routine control, was evaluated. EOs belonging to Mentha, Cymbopogon, and Lavandula families and to S. rosmarinus and T. vulgaris species were analyzed, and the performance of several untargeted approaches, based on the synergistic combination of ATR-FTIR and Partial Least Squares Discriminant Analysis (PLS-DA), was tested to classify the species and chemotypes. Different spectra pre-processing methods were employed, and the robustness of the built models was tested by means of a Receiver Operating Characteristic (ROC) curve and random permutations test. The application of these approaches revealed fruitful results in terms of sensitivity and specificity, highlighting the potentiality of ATR-FTIR and chemometrics techniques to be used as a sensitive, cost-effective, and rapid tool to differentiate EO samples according to their species and chemotype.

Natural volatile oils derived from herbal medicines: A promising therapy way for treating depressive disorder.

Depression is a common global mental disorder that seriously harms human physical and mental health. With the development of society, the increase of pressure and the role of various other factors make the incidence of depression increase year by year. However, there is a lack of drugs that have a fast onset, significant effects, and few side effects. Some volatile oils from traditional natural herbal medicines are usually used to relieve depression and calm emotions, such as Lavender essential oil and Acorus tatarinowii essential oil. It was reported that these volatile oils, are easy to enter the brain through the blood-brain barrier and have good antidepressant effects with little toxicity and side effects. In this review, we summarized the classification of depression, and listed the history of using volatile oils to fight depression in some countries. Importantly, we summarized the anti-depressant natural volatile oils and their monomers from herbal medicine, discussed the anti-depressive mechanisms of the volatile oils from natural medicine. The volatile oils of natural medicine and antidepressant drugs were compared and analyzed, and the application of volatile oils was explained from the clinical use and administration routes. This review would be helpful for the development of potential anti-depressant medicine and provide new alternative treatments for depressive disorders.

Essential Oil Profile and Yield of Corolla, Calyx, Leaf, and Whole Flowering Top of Cultivated Lavandula angustifolia Mill. (Lamiaceae) from Utah.

Lavandula angustifolia Mill. (lavender) is an essential-oil-bearing plant in the Lamiaceae family. Volatile oil produced through the steam distillation of lavender was examined to establish the essential oil yield and aromatic profile from each portion of the plant-namely, the corolla, calyx, leaf, and whole flowering top. The resulting essential oils were analyzed by GC-FID and GC-MS. The different plant parts generally shared similar compounds but in varying relative percentages. Aromatic profiles of the whole flowering top and calyx were similar, with prominent compounds being linalool acetate (34.3%, 32.0%), linalool (26.5%, 32.9%), lavandulyl acetate (5.6%, 4.9%), terpinen-4-ol (5.3%, 7.0%), and (Z)-ß-ocimene (4.5%, 5.4%), respectively. Aromatic profiles for the corolla and leaf were unique. Prominent aromatic compounds of the corolla included linalool acetate (18.4%), linalool (10.8%), epi-α-cadinol (10.0%), borneol (7.3%), and lavandulyl acetate (6.3%). Prominent aromatic compounds of the leaf included epi-α-cadinol (19.8%), γ-cadinene (11.0%), borneol (6.0%), caryophyllene oxide (4.9%), and bornyl acetate (4.8%). Complete profiles and essential oil yields of corolla, calyx, leaf, and whole flowering top were established. This study establishes the influence the corolla, calyx, and leaf exert on the aromatic profile of the whole flowering top and provides insight into authentication of lavender essential oil.

Chemical Variability and In Vitro Anti-Inflammatory Activity of Leaf Essential Oil from Ivorian Isolona dewevrei (De Wild. & T. Durand) Engl. & Diels.

The chemical variability and the in vitro anti-inflammatory activity of the leaf essential oil from Ivorian Isolona dewevrei were investigated for the first time. Forty-seven oil samples were analyzed using a combination of CC, GC(RI), GC-MS and 13C-NMR, thus leading to the identification of 113 constituents (90.8-98.9%). As the main components varied drastically from sample to sample, the 47 oil compositions were submitted to hierarchical cluster and principal components analyses. Three distinct groups, each divided into two subgroups, were evidenced. Subgroup I-A was dominated by (Z)-ß-ocimene, ß-eudesmol, germacrene D and (E)-ß-ocimene, while (10ßH)-1ß,8ß-oxido-cadina-4-ene, santalenone, trans-α-bergamotene and trans-ß-bergamotene were the main compounds of Subgroup I-B. The prevalent constituents of Subgroup II-A were germacrene B, (E)-ß-caryophyllene, (5αH,10ßMe)-6,12-oxido-elema-1,3,6,11(12)-tetraene and γ-elemene. Subgroup II-B displayed germacrene B, germacrene D and (Z)-ß-ocimene as the majority compounds. Germacrene D was the most abundant constituent of Group III, followed in Subgroup III-A by (E)-ß-caryophyllene, (10ßH)-1ß,8ß-oxido-cadina-4-ene, germacrene D-8-one, and then in Subgroup III-B by (Z)-ß-ocimene and (E)-ß-ocimene. The observed qualitative and quantitative chemical variability was probably due to combined factors, mostly phenology and season, then harvest site to a lesser extent. The lipoxygenase inhibition by a leaf oil sample was also evaluated. The oil IC50 (0.020 ± 0.005 mg/mL) was slightly higher than the non-competitive lipoxygenase inhibitor NDGA IC50 (0.013 ± 0.003 mg/mL), suggesting a significant in vitro anti-inflammatory potential.

Growth and production of volatile compounds of yarrow (Achillea millefolium L.) under different irrigation depths.

Yarrow (Achillea millefolium L., Asteraceae) is an important medicinal plant used worldwide for its medicinal properties such as the analgesic, antioxidant and anti-inflammatory ones. The aim of this study was to evaluate the growth and production of photosynthetic pigments and of volatile constituents of Achillea millefolium L. under different irrigation depths. The treatments were the application of 55, 110, 220, 440 and 880 mm of water for a period of 110 days. Data were submitted to polynomial regression analysis at 5% probability, while the volatile constituents were analyzed by standard deviation. Different irrigation depths provided quadratic growth responses being the highest dry matter production at the depth of 440 mm. The contents of chlorophyll a, b, total and carotenoids were higher at the lower depth tested (55 mm). The major volatile compounds identified were sabinene, 1,8-cineol, borneol and ß-caryophyllene. Increased water availability reduced the complexity of the volatile fraction of essential oil. Thus, it is recommended that the species be cultivated at 440 mm irrigation depth to have a higher production of dry matter and lower variation in the volatile profile of the essential oil.

Genetic diversity and metabolic profile of Salvia officinalis populations: implications for advanced breeding strategies.

MAIN CONCLUSION: As a result of this work, we were able to characterize seven indigenous to Greece Salvia officinalis populations using genetic and metabolomic tools. These tools can be used to select the most promising genotypes, capable to design future breeding programs for high valuable varieties. An initial investigation was carried out to compare the genetic and metabolic diversity in S. officinalis grown in Greece and to discern the relationship between the two sets of data. Analysis of inter-simple sequence repeats (ISSR) revealed significant genetic differences among seven sage populations, which were grouped into three main clusters according to an UPGMA ISSR data-based dendrogram and Principle Coordinate Analysis. 80 loci were scored of which up to 90% were polymorphic at species level. According to the composition of their essential oil, the populations were classified into two chemotypes: 1.8 cineole/α-thujone and α-thujone/1.8 cineole. Additionally, a targeted ultra performance liquid chromatography (UPLC-MS/MS) method was used to qualify and quantify phenolic compounds in methanolic extracts of the seven sage genotypes according to which they were districted in six clusters among the sage populations. The main compounds characterizing the seven genotypes were rosmarinic acid and carnosol, followed by apigenin-7-O-glucoside (Ap7glc), and luteolin-7-O-glucoside (Lu7glc). The correlation between matrices obtained from ISSR data and metabolic profiles was non-significant. However, based on the differences in metabolic fingerprint, we aimed to define populations using as main selection criteria the high polyphenol content and desired essential oil composition, using state to the art analytical tools for the identification of parent lines for breeding programs.

Contribution to a Taxonomic Revision of the Sicilian Helichrysum Taxa by PCA Analysis of Their Essential-Oil Compositions.

The chemical profile of the essential oils in ten populations of the genus Helichrysum Mill. (Asteraceae), collected in the loci classici of the nomenclatural types of the taxa endemic to Sicily, were analyzed. Our results confirm that the analysis of secondary metabolites can be used to fingerprint wild populations of Helichrysum, the chemical profiles being coherent with the systematic arrangement of the investigated populations in three main clusters, referring to the aggregates of H. stoechas, H. rupestre, and H. italicum, all belonging to the section Stoechadina. The correct nomenclatural designation of the investigated populations is discussed and the following two new combinations are proposed: Helichrysum preslianum subsp. compactum (Guss.) Maggio, Bruno, Guarino, Senatore & Ilardi and Helichrysum panormitanum subsp. latifolium Maggio, Bruno, Guarino, Senatore & Ilardi.

Volatile compounds in Thymus sect. Teucrioides (Lamiaceae): intraspecific and interspecific diversity, chemotaxonomic significance and exploitation potential.

Thymus sect. Teucrioides comprises three species, namely, T. hartvigii, T. leucospermus, and T. teucrioides, distributed in Greece and Albania. The volatile constituents of all species of the section were obtained by hydrodistillation and investigated by GC-FID and GC/MS analyses. Twenty populations were sampled and a total of 103 compounds were identified, representing 98.0-99.9% of the oil compositions. The oils were mainly characterized by high contents of monoterpene hydrocarbons (42.7-92.4%), with the exception of three oils for which oxygenated monoterpenes were the dominating constituents, viz., that of T. hartvigii ssp. macrocalyx, with linalool as main compound (89.2±0.5%), and those of T. hartvigii ssp. hartvigii and of one population of T. teucrioides ssp. candilicus, containing thymol as major component (46.4±3.1 and 38.2±3.9%, resp.). The most common compound in the oils of the 20 populations of the section was p-cymene. Considerable variation was detected within and among populations, and seven chemotypes were distinguished, i.e., p-cymene, linalool, p-cymene/thymol, p-cymene/γ-terpinene, p-cymene/borneol, p-cymene/γ-terpinene/borneol, and p-cymene/linalool chemotypes. Different chemotypes may exist in the same population. Multivariate statistical analyses enabled the segregation of the oils within Thymus sect. Teucrioides into two groups, one consisting of the three subspecies of T. teucrioides and the second comprising the species T. hartvigii and T. leucospermus. A linalool-rich chemotype, unique within the section, distinguished the oil of T. hartvigii ssp. macrocalyx from all other oils. The high oil content of p-cymene and the preference for serpentine substrates render T. teucrioides species promising for future exploitation.

[Application characteristics and situation analysis of volatile oils in database of Chinese patent medicine].

Aromatic traditional Chinese medicines have a long history in China, with wide varieties. Volatile oils are active ingredients extracted from aromatic herbal medicines, which usually contain tens or hundreds of ingredients, with many biological activities. Therefore, volatile oils are often used in combined prescriptions and made into various efficient preparations for oral administration or external use. Based on the sources from the database of Newly Edited National Chinese Traditional Patent Medicines (the second edition), the author selected 266 Chinese patent medicines containing volatile oils in this paper, and then established an information sheet covering such items as name, dosage, dosage form, specification and usage, and main functions. Subsequently, on the basis of the multidisciplinary knowledge of pharmaceutics, traditional Chinese pharmacology and basic theory of traditional Chinese medicine, efforts were also made in the statistics of the dosage form and usage, variety of volatile oils and main functions, as well as the status analysis on volatile oils in terms of the dosage form development, prescription development, drug instruction and quality control, in order to lay a foundation for the further exploration of the market development situations of volatile oils and the future development orientation.

Essential oils of Micromeria dalmatica Benth., a Balkan endemic species of section Pseudomelissa.

The essential oils of 13 Greek populations of Micromeria dalmatica, a Balkan endemic species and member of the section Pseudomelissa, were examined for the first time. Among the studied populations, two main oil types could be distinguished. Type I was found to be rich in ß-pinene, limonene, and germacrene D (accounting for 55.6-70.2% of the total oil), and Type II was characterized by the preponderance of p-menthane compounds (accounting for 64.2-89.9% of the oil). The latter oil type could be further divided into two subtypes, one comprising oils with predominance of piperitenone and piperitenone oxide and another composed of oils containing high proportions of pulegone, menthone, and isomenthone. The abundance of p-menthane compounds is a common feature of the oils of all members of the section Pseudomelissa studied to date. However, the existence of oils of Type I has not been previously reported for M. dalmatica, neither for other members of the section Pseudomelissa.

Chemical variability of the leaf essential oil of Xylopia aethiopica (Dunal) A.Rich. from Côte d'Ivoire.

The chemical composition of 48 essential-oil samples isolated from the leaves of Xylopia aethiopica harvested in six Ivoirian forests was investigated by GC-FID and (13) C-NMR analyses. In total, 23 components accounting for 82.5-96.1% of the oil composition were identified. The composition was dominated by the monoterpene hydrocarbons ß-pinene (up to 61.1%) and α-pinene (up to 18.6%) and the sesquiterpene hydrocarbon germacrene D (up to 28.7%). Hierarchical cluster and principal component analyses allowed the distinction of two groups on the basis of the ß-pinene and germacrene D contents. The chemical composition of the oils of Group I (38 oil samples) was clearly dominated by ß-pinene, while those of Group II (10 samples) were characterized by the association of ß-pinene and germacrene D. The leaves collected in the four inland forests produced ß-pinene-rich oils (Group I), while the oil samples belonging to Group II were isolated from leaves harvested in forests located near the littoral.

Volatile secondary metabolites of Micromeria dalmatica Benth. (Lamiaceae): biosynthetical and chemotaxonomical aspects.

Analysis by GC and GC/MS of the essential oil obtained from above-ground parts of Micromeria dalmatica Benth. allowed the identification of 116 components, comprising 93.6% of the total oil composition. The major compounds are 3-oxygenated p-menthane monoterpenes and were identified as pulegone (29.6%), menthone (11.7%), and piperitenone (10.8%). The chemical composition of this and additional 30 oils obtained from selected Micromeria Benth. taxa were compared by using multivariate statistical analysis (agglomerative hierarchical cluster analysis and principal component analysis (PCA)). The results of statistical analyses, as well as the domination of different concurrent p-menthane-skeleton-type monoterpene biosynthetical sub-branches in the compared M. dalmatica samples, implied the occurrence of at least two different chemotypes of the mentioned species.

Terpene chemodiversity of relict conifers Picea omorika, Pinus heldreichii, and Pinus peuce, endemic to Balkan.

Terpenes are often used as ecological and chemotaxonomic markers of plant species, as well as for estimation of geographic variability. Essential oils of relic and Balkan endemic/subendemic conifers, Picea omorika, Pinus heldreichii, and P. peuce, in central part of Balkan Peninsula (Serbia and Montenegro), on the level of terpene classes and common terpene compounds were investigated. In finding terpene combinations, which could show the best diversity between species and their natural populations, several statistical methods were applied. Apart from the content of different terpene classes (P. omorika has the most abundant O-containing monoterpenes and sesquiterpenes; P. heldreichii and P. peuce have the largest abundance of sesquiterpene and monoterpene hydrocarbons, resp.), the species are clearly separated according to terpene profile with 22 common compounds. But, divergences in their populations were established only in combination of several compounds (specific for each species), and they were found to be the results of geomorphologic, climatic, and genetic factors. We found similarities between investigated species and some taxa from literature with respect to terpene composition, possibly due to hybridization and phylogenetic relations. Obtained results are also important regarding to chemotaxonomy, biogeography, phylogeny, and evolution of these taxa.

Headspace gas chromatography-mass spectrometry in the analysis of lavender's essential oil: Optimization by response surface methodology.

A static headspace gas chromatography - mass spectrometry (HS-GC/MS) method was developed and optimized with the aim to be applied in the analysis of lavender essential oil. To obtain a comprehensive profile of the essential oil, the optimum HS-GC/MS method parameters were selected based on a Design of Experiments (DοE) process. Plackett-Burman experimental design was applied by utilizing seven parameters of the HS injection system. Incubation equilibration temperature and time, agitator's vortex speed, post injection dwell time, inlet temperature, split ratio and injection flow rate were screened to select the optimum conditions on the basis of the number and the intensity of the identified compounds. Other parameters, such as sample volume and dilution solvent ratio, were also examined to achieve a comprehensive profile in a chromatographic run of 55 min. With the obtained optimum method, more than 40 volatile compounds were identified in lavender's essential oils from different geographical regions in Greece. The method can be utilized for the quality assessment of lavender's essential oil and provide information on its characteristic aroma and discrimination among species based on the acquired GC-MS profiles.

Essential oils and their components in combating fungal pathogens of animal and human skin.

OBJECTIVE: The most common problems of modern medicine include fungal infections of the skin and its appendages caused by dermatomycetes, yeast-like fungi and moulds. Due to toxicity of pharmacological fungicides and promotion of ecology, natural substances with high antifungal properties are sought. Essential oils and their components show potential in this regard. MATERIAL AND METHOD: Fourteen commercial essential oils were tested for antifungal activity. The study were carried out by agar dilution method against the following fungal species: Microsporum gypseum, Microsporum canis, Trichophyton mentagrophytes, Trichophyton violaceum, Aspergillus niger, Scopulariopsis brevicaulis and (IZ 1) dog skin isolate. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) values were determined for all tested oils and individual components of oils with the highest activity against the tested fungi. GC-MS analysis was performed for the most active oils. RESULTS: Cinnamon, thyme, clove, geranium and manuka oils were most active against the test fungi. The MIC values in the tested oils were in the range of 0.5->10µg/µl and the MFC amounted to 1.25->10µg/µl. Activity of individual components against the tested fungi strains was selective and clearly higher against dermatomycetes fungi and IZ 1 isolate. The strongest effect on dermatomycetes fungi was of cinnamaldehyde, thymol and carvacrol-with the values of 0.039-1.25µg/µl (MIC) and 0.078-1.25µg/µl (MFC). In turn, the highest activity against IZ 1 isolate was of cinnamaldehyde, eugenol, carvacrol, geraniol and thymol. Their MIC and MFC was 0.313-1.25µg/µl. CONCLUSION: The results prove the applicability of selected oils and their components as alternatives to synthetic agents in combating fungal dermatoses in animals and humans.

Carvacrol, Thymol, and Garlic Essential Oil Promote Skin Innate Immunity in Gilthead Seabream (Sparus aurata) Through the Multifactorial Modulation of the Secretory Pathway and Enhancement of Mucus Protective Capacity.

One of the main targets for the use of phytogenics in aquafeeds is the mucosal tissues as they constitute a physical and biochemical shield against environmental and pathogenic threats, comprising elements from both the innate and acquired immunity. In the present study, the modulation of the skin transcriptional immune response, the bacterial growth capacity in skin mucus, and the overall health condition of gilthead seabream (Sparus aurata) juveniles fed a dietary supplementation of garlic essential oil, carvacrol, and thymol were assessed. The enrichment analysis of the skin transcriptional profile of fish fed the phytogenic-supplemented diet revealed the regulation of genes associated to cellular components involved in the secretory pathway, suggesting the stimulation, and recruitment of phagocytic cells. Genes recognized by their involvement in non-specific immune response were also identified in the analysis. The promotion of the secretion of non-specific immune molecules into the skin mucus was proposed to be involved in the in vitro decreased growth capacity of pathogenic bacteria in the mucus of fish fed the phytogenic-supplemented diet. Although the mucus antioxidant capacity was not affected by the phytogenics supplementation, the regulation of genes coding for oxidative stress enzymes suggested the reduction of the skin oxidative stress. Additionally, the decreased levels of cortisol in mucus indicated a reduction in the fish allostatic load due to the properties of the tested additive. Altogether, the dietary garlic, carvacrol, and thymol appear to promote the gilthead seabream skin innate immunity and the mucus protective capacity, decreasing its susceptibility to be colonized by pathogenic bacteria.

Essential oil composition of stems and fruits of Caralluma europaea N.E.Br. (Apocynaceae).

The essential oil of the stems and fruits of Caralluma europaea (Guss.) N.E.Br. (Apocynaceae) from Lampedusa Island has been obtained by hydrodistillation and its composition analyzed. The analyses allowed the identification and quantification of 74 volatile compounds, of which 16 were aromatic and 58 non-aromatic. Stems and fruits contained 1.4% and 2.7% of aromatic compounds respectively, while non-aromatic were 88.3% and 88.8%. Non-aromatic hydrocarbons were the most abundant compounds in both organs, followed by fatty acids. Data showed differences in the profiles between stems and fruits which shared only eighteen compounds; stems accounted for 38 compounds while fruits for 53. Fruits showed a higher diversity especially in aromatic compounds with twelve versus four in stems. Among the volatiles identified in stems and fruits of C. europaea 26 are present in other taxa of Apocynaceae, 52 are semiochemicals for many insects, and 21 have antimicrobial activity. The possible ecological role of the volatiles found is briefly discussed.

[Repellent activity of plant essential oils against bites of Lutzomyia migonei (Diptera: Psychodidae)]. / Actividad repelente de aceites esenciales contra las picaduras de Lutzomyia migonei (Diptera: Psychodidae).

Natural repellents from plant extracts have demonstrated good efficacy against bites of some insect species. The present study evaluated the repellent effect of essential oils extracted from 8 plants species against bites of Lutzomyia migonei, the Leishmania vector. The essential oils were extracted by steam destillation in Clevenger chamber, from the following plants: Hyptis suaveolens, Pimenta racemosa, Piper marginatum, Monticalia imbricatifolia, Pseudognaphalium caeruleocanum, Espeletia shultzii, Plecthranthus amboinicus and Cinnamomun zeylanicum. Repellency tests were performed under laboratory conditions by the human hand method in cage assays, using female colonies of L. migonei. The more effective oils were tested at variable concentrations on different volunteers. The protection percentage and time were calculated. The results showed what oils of P. caeruleocanum and C. zeylanicum were the most effective. Although P. amboinicus oil also had repellent effect showed an irritant effect. The oils P. marginatum, H. suaveolens and P. racemosa showed no repellent effect, while the rest of oil extracts showed significant repellency in variable degrees. P. caeruleocanum and C. zeylanicum oils provided the 95% protection against bites of L. migonei for 3 h. The P. caeruleocanum oil showed the greatest protection time, with a mean over 4h and 3h at concentrations of 50% and 10% respectively. The results suggest that the P. caeruleocanum oil could represent a potential natural repellent against Leishmania vectors.

Partitioning the relative contributions of inorganic plant composition and soil characteristics to the quality of Helichrysum italicum subsp. italicum (Roth) G. Don fil. essential oil.

Composition of Helichrysum italicum subsp. italicum essential oil showed chemical variability according to vegetation cycle, environment, and geographic origins. In the present work, 48 individuals of this plant at different development stages and the corresponding root soils were sampled: i) 28 volatile components were identified and measured in essential oil by using GC and GC/MS; ii) ten elements from plants and soils have been estimated using colorimetry in continuous flux, flame atomic absorption spectrometry, or emission spectrometry (FAAS/FAES); iii) texture and acidity (real and potential) of soil samples were also reported. Relationships between the essential-oil composition, the inorganic plant composition, and the soil characteristics (inorganic composition, texture, and acidity) have been established using multivariate analysis such as Principal Component Analysis (PCA) and partial Redundancy Analysis (RDA). This study demonstrates a high level of intraspecific differences in oil composition due to environmental factors and, more particularly, soil characteristics.

Chemical composition of the essential oils from the aerial parts of two Malagasy endemic species (Apiaceae): Billburttia capensoides Sales & Hedge and Billburttia vaginoides Sales & Hedge.

The chemical composition of twenty-five essential oil samples from the aerial parts of two Malagasy endemic species Billburttia capensoides Sales & Hedge and B. vaginoides Sales & Hedge, were investigated for the first time. Based on chromatographic profiles, three selected samples were investigated using GC(RI), GC-MS and 13C NMR. The content of the main components varied drastically from sample to sample: p-mentha-1,3,8-triene (0.2-52.7%), terpinolene (2.8-40.7%) and dill apiole (0.0-22.2%). Statistical analysis of the 25 oil compositions allowed the distinction of two well-differentiated groups. Samples of group I contained mainly p-mentha-1,3,8-triene while the Group II was dominated by terpinolene and dill apiole.