Hydrodistilled Essential Oil Research Articles

Characterization of Essential Oils of the Leaves and Fruits of Adenanthera pavonina L. by GC/MS

The characterization of the hydro distilled essential oils of the leaves and fruits of A. pavonina, growing in Egypt, was analyzed by gas chromatography-mass spectrometry (GC-MS) using Rtx-5MS fused bonded column. The results revealed the identification of 27 compounds in the leaves and fruits oils. The leaves oil consisted mainly of monoterpene hydrocarbons (68.01%). The major compounds from the leaves oil were sabinene, accounting for 27.9% of the oil content followed by D-limonene (14.79%), 3-carene (8.46%), caryophyllene (6.57%), and γ-terpinene (6.16%). The fruits oil consisted mainly of sesquiterpene hydrocarbons (62.8%). The major compounds were β-element (39.6%) followed by sabinene (11.7%) and caryophyllene (11.38%). These findings will assist in the differentiation between the volatile oils from different parts of A. pavonina is a step to establish the first monograph for this plant. This study summarized the chemical profiles of A. pavonina leaves and fruits essential oils growing in Egypt for the first time. The leaves essential oils of A. pavonina demonstrated different composition compared with the oils from the fruits.

Chemical Composition and Antimicrobial Activity of Essential Oil from the Stem of Magnolia coco (Lour.) DC. from Vietnam

The chemical constituents and antimicrobial activity of hydro distilled essential oil from the stem of Magnolia coco (Lour.) DC. were reported. The constituents of the essential oil were determined using gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC/MS) while the antimicrobial action was evaluated by using microdilution broth susceptibility assay. The major com- pounds in the essential oil were sabinene (31.9 %) and β-pinene (11.8 %). The antimicrobial study depicted by minimum inhibitory concentration (MIC) showed that the essential oil was active against Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 14579 and Candida albicans ATCC 10231 with MIC value of 16.0 μg/mL as well as Enterococcus faecalis ATCC 299212 and Pseudomonas aeruginosa ATCC 27853 with MIC value of 32.0 μg/mL. The obtained IC50 values of the antimicrobial efficacy were 7.89, 7.66, 8.33, 16.44, and 15.67 μg/mL respectively. This report, the first of its kind, indicates the potential of M. coco stem essential oil as a source of anti-microbial principle.

A further study of the kinetics and optimization of the essential oil hydrodistillation from lavender flowers

Abstract Hydrodistillation has commonly been used to recover essential oil from various plant materials, including lavender (Lavandula officinalis) flowers. The main objectives of the present study were to model the kinetics of the lavender essential oil (LEO) hydrodistillation using a phenomenological model, to evaluate the statistical significance of the hydromodule and hydrodistillation time on LEO yield combining a 42 full factorial design with the response surface methodology, to model statistically LEO yield by multiple non-linear regression and to determine the optimal process conditions that provided the maximum LEO yield. The fast-essential oil distillation (washing stage) in the initial period and the slow diffusion stage until the saturation occurring simultaneously were observed, justifying the use of the phenomenological model. With increasing the hydromodule, the saturation LEO yield and the washable fraction of the LEO decreased while the washing and diffusion rate constants increased. Knowledge of the LEO oil yield and the hydrodistillation kinetics is important from the techno-economical point of view.

Chemical composition of essential oils of berries of Juniperus macrocarpa Sibth. & Sm. from Turkey

The chemical compositions of the hydrodistilled essential oils of ripen and unripen berries of Juniperus macrocarpa Sibth. & Sm. (Jom) were analysed by GC-MS. Thirty eight compounds were identified from unripe berries, accounting for approximately 0.03% oil yield (v/w dried weight) and 16 compounds from ripe berries with 0.003% oil yield (v/w dried weight). While the major compounds in the essential oils of unripe berries were α-cedrol (30.04%), α-pinene (18.81%), germacrene-D (14.58%), δ-cadinene (3.80%), γ-cadinene (2.29%), α-humulene (2.29%), γ-muurolene (2.27%), epi-bicyclosesquiphellandrene (1.69%), transcaryophyllene (1.68%), valencen (1.53 %) and α-cedrol (49.87%), β- caryophyllene (29.57%), α-cedrene (4.75%), γ-muurolene (2.54%), carvacrol methyl ether (1.4%), trans-pinocarveol (1.2%), α-pinene (1.13%), α-muurolene (1.04%) were identified from ripe berries and their mean percentage varied according to their phenological stage. Both qualitative and quantitative differences between ripe and unripe berries of the plant were observed.

Phytotoxic and Antimicrobial Activities of Teucrium polium and Thymus decussatus Essential Oils Extracted Using Hydrodistillation and Microwave-Assisted Techniques.

Essential oils (EOs) have been described as promising eco-friendly secondary products of aromatic plants with several biological activities. The present study aimed to characterize the chemical composition and explore phytotoxic and antimicrobial activities of Teucrium polium and Thymus decussatus EOs extracted using hydrodistillation (HD) and microwave-assisted extraction (MAE) methods. Twenty-seven and twenty-eight compounds were identified from HD and MAE extracted EOs of T. polium, respectively. The oxygenated sesquiterpenes (57.68%) were characterized as the main components of the hydrodistilled EO with a prominence of 6-epi-shyobunol (33.00%), while sesquiterpene hydrocarbons (54.48%) were the main components of the MAE method, with a prominence of delta-cadinene (25.13%). Eighteen and nineteen compounds, were characterized in T. decussatus EOs extracted using HD and MAE methods, respectively, and oxygenated monoterpenes represented the main components of both EOs with carvacrol (94.40% and 75.91%, respectively) as the main compound. The EOs extracted using the MAE method were slightly more phytotoxic than those extracted using the HD method. The T. decussatus EO extracted using the MAE method showed a higher inhibitory effect than T. polium by 16-, 32-, and 24-fold, regarding seed germination, shoot, and root growth of lettuce, respectively. Moreover, EOs extracted by HD method showed a similar pattern with 16-, 28-, and 14-fold effects. Both T. decussatus EOs exhibited potent inhibitory effect against all tested bacteria with an inhibition zone of 34–39 mm and the lowest minimum inhibitory concentration (MIC) of 0.49, 0.98, and 1.95 μg/mL against Aspergillus niger, Escherichia coli, and Staphylococcus aureus, respectively. However, the EOs of T. polium showed weak antibacterial activity and no antifungal effect. Further studies are needed for the characterization of bioactive major compounds, either singular or synergistic, at field scale and to determine their modes of action and safety.

Chemical Composition and Insecticidal Activities of the Essential Oil from Caryopteris incana (Thunb. ex Hout.) Miq. Aerial Parts

Hydro distilled essential oil of the aerial parts of Caryopteris incana harvested from Zhejiang province, China was analyzed by gas chromatography-mass spectrometry (GC/MS). Thirty-nine components were identified and the principal compounds in the essential oil were 4-terpineol (13.5 %), 1-octen-3-ol (12.3 %), myrtenol (8.8 %), and α-cadinol (8.5 %). Three active compounds (α-cadinol, myrtenol, 4-terpineol) were isolated and identified from the essential oil based on bioactivity-directed fractionation. The essential oil and its isolates, myrtenol and 4-terpineol exhibited fumigant activity against the red flour beetles, Tribolium castaneum with LC50 values of 2.2, 5.6, and 0.5 mg/l, respectively. The essential oil and the three constituents, α-cadinol, myrtenol, and 4-terpineol also displayed contact toxicity against T. castaneum with LC50 values of 30.7, 123.4, 29.7, and 21.3 μg/adult, respectively. The study indicates that the essential oil of C. incana and its isolated constituents have the potential to be developed to natural insecticides for the control of grain storage insects.

Aroma volatiles, phenolic profile and hypoglycaemic activity of Ajuga iva L.

The phenolic profile and aroma volatiles of aerial parts of wild Ajuga iva L., collected from the Tunisian Desert, have been investigated. Sixteen phenolic compounds in the methalonic extract were identified including 7 phenolic acids and 9 flavonoids. The major compounds were cirilisiol (32 ± 2%), chlorogenic acid (2.4 ± 0.1%), cirsilineol (1.6 ± 1%), hyperoside (1.7 ± 0.1%), syringic acid (0.7 ± 0.1%), and quinic acid (0.6 ± 0.1%). The chromatographic analysis showed three volatile compounds detected in hydrodistilled essential oil (HDEO), representing 99% of the total oil, while solid-phase microextraction (SPME) showed 6 components constituting 98% of the volatile material. The results showed that monoterpenes β-pinene (70 ± 5%) and α-pinene (20 ± 2%) were predominant in SPME and β-pinene (59 ± 4%) and limonene (26 ± 4%) were predominant in HDEO. Furthermore, the in vivo anti-diabetic effect of the extract of A. iva aerial parts was monitored. As a supplement at 50% of extract seemed to be beneficial for significantly reducing hyperglycemia (~4 fold), after 8 days in rats.

GC-MS Analysis of Volatile Organic Constituents of Traditionally Used Medicinal Plants from the Western Ghats of India: Blumea lanceolaria (Roxb.) Druce., Heliotropium indicum L. and Triumfetta rhomboidea Jacq.

The present study explored essential oil constituents of the medicinal plants used in traditional medicine and growing wild in the Western Ghats region of India, which is one of the 34 global biodiversity hotspots. The hydro distilled essential oils of Blumea lanceolaria (Roxb.) Druce. (Asteraceae), Heliotropium indicum L. (Boraginaceae) and Triumfetta rhomboidea Jacq. (Tiliaceae) were analyzed using gas chromatography equipped with a flame ionization detector (GC-FID) and gas chromatography coupled with mass spectroscopy (GC/MS). Thirty-six, twenty-seven, and fifty constituents were identified from the essential oils of B. lanceolaria, H. indicum, and T. rhomboidea, respectively comprising 97.3% (B. lanceolaria), 98.2% (H. indicum) and 97.6% (T. rhomboidea) of the total oil constituents. The major compounds of B. lanceolaria, H. indicum, and T. rhomboidea were identified as β-pinene (82.3%), methyl salicylate (54.3%), and β-caryophyllene (28.9%), respectively. The essential of B. lanceolaria was found to be rich in monoterpene hydrocarbons (90.4%), while H. indicum and T. rhomboidea oils were rich in phenyl derivative (72.8%) and sesquiterpene hydrocarbons (69.5%) type constituents, respectively. The common compounds viz., terpin-4-ol and eugenol were identified in the essential oils of B. lanceolaria, H. indicum, and T. rhomboidea.

Chemical composition and in vitro activity of Origanum vulgare L., Satureja hortensis L., Thymus serpyllum L. and Thymus vulgaris L. essential oils towards oral isolates of Candida albicans and Candida glabrata

AbstractThe purpose of this research was to investigate the chemical composition of essential oils (EOs) from:Origanum vulgareL.,Satureja hortensisL.,Thymus serpyllumL. andThymus vulgarisL. (Lamiaceae) cultivated in Poland, and to study their antifungal activity towards clinical isolates of oralCandidaspp. The hydrodistilled essential oils were analyzed using the GC-MS method. The antifungal activity was evaluatedin vitroagainst oral isolates and reference strains ofCandida albicansandC. glabrata, using the broth microdilution method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) guidelines, allowing for estimation of minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). GC-MS analysis revealed that carvacrol was the main EO compound in oregano and summer savory, while thymol and linalool were the major ingredients of thyme and wild thyme oils, respectively. The EOs possessed fungicidal activity againstC. albicansandC. glabrata, including oral isolates, with MIC = 125 – 2000 mg/L, MFC = 250 – 4000 mg/L and MFC/MIC = 1 – 4, depending on the yeast and plant species. The most active was thyme oil – with MIC = 125 – 500 mg/L, MFC = 250 – 500 mg/L and MFC/MIC = 1 – 2.

Effect of nanoencapsulation on volatile constituents, and antioxidant and anticancer activities of Algerian Origanum glandulosum Desf. essential oil

Nanoencapsulation is an attractive novel technique used for incorporating essential oils in food preparations and pharmaceutical formulae. This study investigated the effect of nanoencapsulation on the composition of volatile compounds, as well as the antioxidant and anticancer activities of hydrodistilled (HD) Origanum glandulosum Desf. Oil, which was encapsulated into nanocapsules via High Speed Homogenization (HSH) and into nanoemulsions through High Pressure Homogenization (HPH). Thirty-two volatile components were identified using Gas Chromatography-Mass Spectrometry analysis (GC-MS) in HD essential oil representing 99.04% of the total oil content. GC-MS analysis showed that the use of HPH to prepare nanoemulsions negatively affected the active compounds present in HD oil, particularly carvacrol and thymol, whereas the use of HSH led to significant quantitative differences in the composition of volatiles between HD oil and nanocapsules but generated the same profile. Consistent with the differences in total phenolics, total flavonoids, and volatiles identified in HD and nanoparticles, HD essential oil exhibited a higher antioxidant activity (IC50 4.22 mg/mL) than nanocapsules (IC50 57.51 mg/mL) and nanoemulsion (IC50 78.50 mg/mL), while nanocapsules showed the strongest cytotoxic effect on liver cancer cell line Hep-G2 (54.93 μg/mL) in comparison to HD oil (73.13 μg/mL) and nanoemulsions (131.6 μg/mL).

Enzyme extraction by lab-scale hydrodistillation of ginger essential oil (Zingiber officinale Roscoe): Chromatographic and micromorphological analyses

The present study assessed the use of crude multi-enzymatic extracts (CME) obtained from solid-state fermentation of cocoa bean shells by Aspergillus oryzae ATCC 10124 in the extraction of ginger essential oils, without the use of additional steps, such as drying, which could cause some compounds to volatilize. A Doehlert matrix was used in the optimization of the enzymatic pre-treatment. Independent variables of temperature, time and the proportion of CME/H2O were evaluated. After enzymatic treatment, extraction occurred by hydrodistillation. The results demonstrated that pre-treatment raised essential oil yields by 47.95 % at 40 °C, 130 min and CME/H2O (75/425 mL mL−1). The composition of the obtained essential oils was identified by gas chromatography mass spectrometry (GC–MS), α-zingiberene (25.20 %) being the main volatile constituent. Degradation of the plant cell wall of the ginger was observed through micromorphological analysis. Application of CME in ginger essential oil extraction eliminated the drying stage that is necessary in this type of plant material, reducing both energy consumption of the process and volatilization of the chemical components.

Zingiber zerumbet Rhizome Essential Oil: Chemical Composition, Antimicrobial and Mosquito Larvicidal Activities

Aims: In the present study, the chemical constituents, antimicrobial and larvicidal potentials of hydrodistilled essential oil from the rhizome of Zingiber zerumbet were evaluated.
 Study Design: The study was designed in different phases which are: collection of mature rhizomes of Zingiber zerumbet, hydrodistillation of essential oils, chemical analysis of the essential oils, determination of the antimicrobial potential and evaluation of larvicidal activity.
 Place and Duration of Study: The study was conducted at School of Natural Science, Vinh University, Vinh City, Nghệ An Province, Vietnam. The duration of the study was between August and December 2018.
 Methodology: The rhizomes of Z. zerumbet were collected from Bến En National Park, Thanh Hóa Province, Vietnam, in August 2018. The air-dry sample was subjected to hydrodistillation process using Clevenger-apparatus to obtained essential oils. We evaluated the larvicidal potential of the oil against Culex quinquefasciatus and Aedes albopictus at 24 h and 48 h according to World Health Organisation protocol. The antimicrobial activity (MIC) was determined by microdilution broth susceptibility assay. Statistical analysis was performed using GraphPad Prism (version 7.02). The LC50 values, LC90 values and 95% confidence limits were obtained by using XLSTAT v. 2018.5.
 Results: The most abundant compound of the essential oil was zerumbone (51.3%). The essential oil showed mortality of 98.3% (24 h) and 100% (48 h) against the Ae. albopictus at concentration of 100 µg/mL. In the same vein, mortality of 100% was displayed against Cx. quinquefasciatus under the tested time and concentration. The essential oil exhibited larvicidal activity towards Cx. quinquefasciatus showing minimum lethal concentrations, LC50, values of 33.28 mg/mL (24 h) and 21.81 mg/mL (48 h). The LC50 values of 55.75 µg/mL and 36.22 µg/mL at 24 h and 48 h respectively were obtained against Ae. albopictus. The result of the antimicrobial test indicated that Z. zerumbet oil inhibited the growth of Aspergillus niger (ATCC 9763) with MIC of 50.0 µg/mL.
 Conclusion: Results demonstrated that the essential oil of Z. zerumbet was effective in the control of tested mosquitoes, Culex quinquefasciatus and Aedes albopictus and the microbe, Aspergillus niger.

The Essential Oil and Hydrolats from Myristica fragrans Seeds with Magnesium Aluminometasilicate as Excipient: Antioxidant, Antibacterial, and Anti-inflammatory Activity.

Nutmeg (Myristica fragrans) essential oil has antimicrobial, antiseptic, antiparasitic, anti-inflammatory, and antioxidant properties. We have recently demonstrated that hydrodistillation of nutmeg essential oil by applying magnesium aluminometasilicate as an excipient significantly increases both the content and amount of bioactive substances in the oil and hydrolats. In this study, we aimed to compare the antioxidant, antimicrobial, and anti-inflammatory activity of hydrolats and essential oil obtained by hydrodistillation in the presence and absence of magnesium aluminometasilicate as an excipient. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method revealed that magnesium aluminometasilicate did not significantly improved antioxidant activity of both essential oil and hydrolat. Antibacterial efficiency was evaluated by monitoring growth of 15 bacterial strains treated by a range of dilutions of the essential oil and the hydrolats. Essential oil with an excipient completely inhibited the growth of E. faecalis, S. mutans (referent), and P. multocida, whereas the pure oil was only efficient against the latter strain. Finally, the anti-inflammatory properties of the substances were assessed in a fibroblast cell culture treated with viral dsRNR mimetic Poly I:C. The essential oil with an excipient protected cells against Poly I:C-induced necrosis more efficiently compared to pure essential oil. Also, both the oil and the hydrolats with aluminometasilicate were more efficient in preventing IL-6 release in the presence of Poly I:C. Our results show that the use of magnesium aluminometasilicate as an excipient might change and in some cases improve the biological activities of nutmeg essential oil and hydrolats.

Optimization of microwave assisted hydrodistillation of essential oil from lemon (Citrus aurantifolia) leaves: Response surface methodology studies

In this paper, microwave-assisted hydro-distillation (MAHD) of essential oils from the lemon (Citrus aurantifolia) leaves was attempted and optimized. Optimization of MAHD was performed by using response surface methodology. In the optimization, selected parameters consisted of water and material ratio, microwave power and extraction time were. It is also indicated that quadratic polynomial model could be employed to optimize the microwave extraction of essential oil from Lemon (Citrus aurantifolia) leaves. The optimal extraction conditions included microwave power of 523.89W, water to material ratio of 3.27 mL/g, and extraction time of 84.47 minutes. The optimized yield (0.76%) approached predicted yield predicted by the model, implying that the model is suitable to predict behavior of the process.

Composition of essential oils and headspace constituents of Artemisia annua L. and A. scoparia Waldst. et Kit.

Headspace volatiles (HS) and hydrodistilled essential oils (EO) of fresh aerial parts of Artemisia annua L. and A. scoparia Waldst. et Kit., were analyzed by GC?MS/FID. Artemisia ketone was found to be the most abundant component among the EO volatiles (55.8 %), as well as among HS (52.1 %) of A. annua. Additionally, in both A. annua samples, EO and HS, ?-pinene (12.7 and 24.2 %, respectively) was found in high percentage. On the other hand, it has been determined that the dominant components of A. scoparia EO and HS were different; in the essential oil capillene (63.8 %) was found as the main constituent, while ?-pinene (26.1 %), (Z)-?-ocimene (23.8 %) and limonene (10.7 %) were the major components among the HS. This is the first report on the composition of HS volatiles of the A. annua and A. scoparia obtained by direct static headspace.

Effect of thyme (Thymus vulgaris L.) essential oil on the quality parameters of stirred yoghurt

Aim Stirred yoghurt was supplemented with thyme essential oil in an attempt to enhance its quality. Materials and methods Thyme (Thymus vulgaris L.) essential oil concentrations (0, 2, 4, 6, 8, and 10 μl/ml) were added to the prepared stirred yoghurt and then examined for microbiological count and mycotoxin quality during storage intervals following the Egyptian standards. Results Samples contained ≥ 6 μl/ml oil showed significant increase (p<0.05) of total viable bacterial count and lactic acid bacteria, whereas mold and yeast count was decreased. Pathogenic bacteria including Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Salmonella spp., Escherichia coli, and coliform group were not detected in any of the samples, which complied with the standards. Moreover, aflatoxin M1 content was reduced by time in direct proportion to the thyme essential oil concentration. Conclusion The bioactive components in thyme act as a natural preservative in stirred yoghurt throughout the storage interval when the yogurt is fortified with hydro-distilled thyme essential oil.

Production of thymol rich extracts from ajwain (Carum copticum L.) and thyme (Thymus vulgaris L.) using supercritical CO2

Supercritical fluid extraction is the most efficient technique to extract flavor and fragrance compounds. This study aims to obtain thymol rich extracts by supercritical fluid CO2-extraction (SFE) of Carum copticum L. (ajwain) and Thymus vulgaris L. (thyme). The yield, chemical constituents analyzed by gas chromatography-mass spectrometry and sensory characteristics of the extracts were compared with their volatile oil counterparts extracted by hydrodistillation (HD). The SFE extracts were obtained at a low temperature (40 °C) and two pressures (10.4 and 16.7 MPa). The yield of the ajwain and thyme extracts reached 2.78 ± 0.04 and 1.28 ± 0.15 % (w/w), respectively, at 16.7 MPa. The SFE successfully increased the thymol relative amount in the ajwain and thyme extracts by 1.7 and 2.49 fold compared with its level in the hydrodistilled essential oils, respectively. The extract obtained at high pressure exhibited the most valued sensory characteristics (spicy, pungent, aromatic/herbaceous, phenol-like and absence of burnt note).

Comparative chemical analysis of volatile compounds of Warionia saharea leaves using hydrodistillation and headspace solid-phase microextraction (HS-SPME)

The objective of this study is to report the comparative chemical analysis of essential oil (EO) obtained by hydrodistillation (HD) and volatile fraction (VF) detected by headspace solid-phase micro-extraction (HS–SPME) isolated from Warionia saharea leaves using Gas Chromatography–Retention Indices (GC-RI) and GC–Mass Spectrometry (GC-MS). 39 volatile compounds identified in hydrodistilled essential oil (HD), representing 92.7% of the total oil, while HS-SPME revealed 25 components constituting 94.1% of the volatile material. The chemical composition of the HS-SPME and HD extract comprised mainly of phenylpropanoids (36.9%) and oxygenated sesquiterpenes (52.8%), respectively. The comparative analysis of two chemoprofiles obtained by two methods shows both qualitative as well as quantitative differences. The current study is the first report involving rapid analysis of volatile components of W. saharea by HS-SPME.

Root Bark of Cordia millenii Essential Oil: Anti-inflammatory and Anti-nociceptive Activities

Aims: Considering the lack of scientific studies focused on the pharmacological activity of Cordia millenii essential oil, this work was designed to evaluate the anti-inflammatory and anti-nociceptive activities of essential oil from the root bark.&#x0D; Study Design: The design of the study include collection of root bark of Cordia millenii, hydrodistillation of essential oil from the plant and evaluation of its anti-inflammatory and anti-nociceptive potentials.&#x0D; Place and Duration of Study: Department of Chemistry, Lagos State University, Nigeria between May 2017 and April 2018.&#x0D; Methodology: The root bark of C. millenii were collected from Ayetoro, Ilesha (7°37¢0N 4°43¢0E), Osun State, Nigeria in June 2017. Essential oils were obtained from the air-dry sample by hydrodistillation procedure in an all glass Clevenger-apparatus. The anti-inflammatory activity was evaluated by carrageenan-induced rat paw edema. The anti-nociceptive action was established from the hot-plate analysis. Statistical analysis was performed using GraphPad Prism (version 7.02).&#x0D; Results: The anti-inflammatory activity of the essential oil was statistically significant (p &lt; 0.001) at 1st and 3rd h for the 100 mg/kg p.o., at 1st-3rd h for the 200/kg mg p.o. and at 3rd and 4th h for the 400 mg/kg p.o. In addition, the 100 mg/kg p.o. showed significant activity (p &lt; 0.01) at 2nd h. Also, the anti-inflammatory activity was significant (p &lt; 0.05) for 100 mg/kg p.o. (4th h), 200 mg/kg p.o. (4th h) and 400 mg/kg p.o (1st h). The essential oil of C. millenii displayed high activity (p &lt;0.001) for all doses in the hot plate anti-nociceptive assay which was time and dose independent.&#x0D; Conclusion: Results demonstrate that the essential oil of C. millenii was effective in the treatment of inflammatory conditions, thereby supporting the traditional use of this herb.

Autopolyploidy effect on morphological variation and essential oil content in Thymus vulgaris L.

Garden thyme (Thymus vulgaris L.) is well known for its aromatic, anti-pathogenic and antioxidant properties and is commonly cultivated for ornamental, culinary and medicinal uses. The aim of this study was to induce ployploidy in T. vulgaris in order to obtain tetraploid plants (2n = 4x = 60) with enhanced morphological traits and increased essential oil content from diploid plants (2n = 2x = 30) using in vitro-induced polyploidization. Nodal segments were treated with four different concentrations of oryzalin (20, 40, 60 and 80 μM) for 24 and 48 h. The ploidy levels of treated and regenerated plants were determined by flow cytometry analysis. The chemical composition of hydrodistilled essential oil was analysed by gas chromatography with quadrupole time-of-flight mass spectrometry systems. Oryzalin treatment resulted in three tetraploid plants at a concentration of 80 μM for 24 h. Obtained tetraploid plants presented several morphological changes (increased plant weight, height, leaf length, breadth and thickness) in comparison to diploid plants. Furthermore, polyploidization demonstrated an increase in the production of secondary metabolites; tetraploids yield of essential oil increased from 0.81 % to 1.19 % and the content of several main components increased, including thymol and carvacrol by 18.01 % and 0.49 %, respectively. The obtained genotype could be implemented as a new variety for commercial production of garden thyme.