Yield and Composition of the Essential Oil of Clinopodium nepeta subsp. spruneri as Affected by Harvest Season and Cultivation Method, i.e., Outdoor, Greenhouse and In Vitro Culture.

Clinopodium nepeta subsp. spruneri is an aromatic herb with a mint-oregano flavor, used in Mediterranean regions in traditional medicine. The aerial parts of the plant are rich in essential oil that has antioxidant, antimicrobial and anti-inflammatory properties as well as insecticidal activity. The aim of our work was to determine the yield and composition of the essential oil of the plant, in relation to the harvest season and cultivation method, i.e., outdoor, greenhouse and in vitro culture, using gas chromatography-mass spectrometry (GC-MS) as an analytical tool. Essential oil yield fluctuated similarly in outdoor and greenhouse plants during the year (0.9-2.6%), with higher percentages (2.1-2.6%) in the hottest periods June-October (flowering stage) and April (vegetative stage), and was similar to the yield in in vitro plants (1.7%). More compounds were identified in the oil of outdoor and greenhouse plants (35) compared to that of in vitro plants (21), while the main compounds were the same, i.e., pulegone (13.0-32.0%, highest in February-April, 15.0% in vitro), piperitenone oxide (3.8-31.8%, lowest in February, 34.2% in vitro), piperitone epoxide (4.6-16.4%, highest in February, 15.5% in vitro), D-limonene (2.1-8.8%, lowest in February, 10.0% in vitro), isomenthone (2.3-23.0%, highest in February, 4.6% in vitro), germacrene D (1.9-6.5% highest in December-April, 2.9% in vitro) and dicyclogermacrene (2.1-5.3%, highest in December-April, 5.2% in vitro). Therefore, greenhouse and in vitro cultures were equally efficient in yielding essential oil and its constituents as outdoor cultivation, while in outdoor and greenhouse cultivations, the harvest season, mainly due to the prevailing ambient temperatures, affected the essential oil yield and its percentage composition.

Chemical Composition of the Essential Oils of Three Popular Sideritis Species Cultivated in Greece Using GC-MS Analysis.

(1) Background: The essential oils (EOs) of Sideritis L. have attracted great interest due to their pharmacological activities and potential applications in the cosmetic and perfume industries. The aim of this work was to study the EO chemical composition of three of the most popular, in Greece, mountain tea species: namely, these include Sideritis scardica, Sideritis raeseri, and Sideritis syriaca. (2) Methods: The EOs were obtained from the aerial parts of three Sideritis species that were cultivated in various regions of Greece by hydrodistillation, and the chemical composition was studied by gas chromatography-mass spectrometry (GC-MS) analysis. (3) Results: The EOs of the Sideritis species-S. scardica (SSC1, SSC2, SSC3), S. raeseri (SR1, SR2, SR3), and S. syriaca (SS1, SS2, SS3)-were analyzed by GC-MS, and they showed both qualitatively and quantitatively high variation in their chemical composition. (4) Conclusions: The EOs of S. scardica and S. raeseri from three different regions of Greece, and the S. syriaca from three different localities of Crete Island in Southern Greece, showed high chemical variability. Although 165 different components were found to be present in the nine samples through GC-MS analysis, only 7 (1-octen-3-ol, linalool, trans-pinocarveol, p-mentha-1,5-dien-8-ol, α-terpineol, myrtenol, and verbenone) were common components in the nine EOs, which were identified to be highly variable in different percentages among the samples. Even the EOs of SS1 and SS2, which were cultivated nearby, showed different GC profiles. The composition variation observed might be attributed to differentiations in the soil and climatic conditions.

Chemical Characterization and Antioxidant Activity of Nine Hypericum Species from Greece.

Hypericum L. comprises about 500 species distributed almost worldwide. Research has mainly focused on H. perforatum with confirmed biological activity on the alleviation of depression symptoms, among others. The compounds responsible for such activity are considered naphthodianthrones and acylphloroglucinols. Other Hypericum species are less studied or not studied, and further research is needed to complete the characterization of the genus. In this study we evaluated the qualitative and quantitative phytochemical profile of nine Hypericum species native to Greece, namely H. perforatum, H. tetrapterum, H. perfoliatum, H. rumeliacum subsp. apollinis, H. vesiculosum, H. cycladicum, H. fragile, H. olympicum and H. delphicum. Qualitative analysis was performed using the LC/Q-TOF/HRMS technique, while quantitative data were calculated with the single point external standard method. Additionally, we estimated the antioxidant activity of the extracts using DPPH and ABTS assays. Three species endemic to Greece (H. cycladicum, H. fragile, H. delphicum) were studied for the first time. Our results indicated that all studied species are rich in secondary metabolites, mainly of the flavonoids family, with strong antioxidant activity.

Origanum majorana Essential Oil-A Review of Its Chemical Profile and Pesticide Activity.

Origanum majorana is a medicinal and aromatic plant that belongs to the Lamiaceae family. It is cultivated in several parts of the world and, due to its splendid aroma and taste, is widely used for culinary purposes and in perfumes. The essential oil of the plant, to which is attributed its aroma, contains many secondary metabolites with valuable biological activity. One of them is the pesticide activity, which has attracted much interest. Given the necessity of replacing synthetic pesticides, essential oils are studied in an attempt to find naturally derived products. Thus, the aim of this review paper is to discuss the chemical profile of O. majorana essential oil and to present data regarding its insecticidal, repellent and fumigant activity. Data were collected from 1992 to 2022. Databases, including PubMed, Google Scholar, ScienceDirect and Scopus, were used for the research, and keywords, including O. majorana, sweet marjoram, essential oil, volatiles, pesticide, insecticide and repellent activity, were used. The results of this review paper indicate that O. majorana essential oil can be an alternative agent to manage pests. However, still, much research should be conducted to evaluate its toxicity against beneficial insects and to ensure its safety for human health.

Synergistic larvicidal and repellent effects of essential oils of three Origanum species on Rhipicephalus annulatus tick.

Ticks are of great economic importance worldwide, both because they represent major obstacles to livestock productivity and because of their ability to transmit diseases to humans and animals. Although synthetic acaricides are the most common method for tick control, their overuse has led to the development of resistance as well as unacceptable residual levels in animal products and in the environment in general. There is therefore an urgent need to identify alternative treatments. Among such alternative approaches for tick control is plant essential oil (EO) therapy. In the present study, we investigated the synergistic effect of EOs of three oregano species-Origanum onites, O. majorana and O. minutiflorum-against Rhipicephalus annulatus larvae. Gas chromatography-mass spectrometry profiles of the three EOs revealed that carvacrol was their major component, with a concentration of 86.2% in O. majorana, 79.1% in O. minutiflorum and 77.4% in O. onites. The results of larvicidal assays revealed that the doses that lead to the death of 50% of the ticks (LC50) were 22.99, 25.08 and 27.06 µL/mL for O. majorana, O. minutiflorum and O. onites EOs, respectively, whereas the doses that lead to the death of 99% (LC99) were 41.26, 43.62 and 48.96 µL/mL. In addition, the LC50 and LC99 of the three oils combined was lower (viz., 4.01 and 6.97 µL/mL) than that of each oil alone. The tested EOs were also able to repel larvae of R. annulatus to varying degrees, with O. onites oil exhibiting the greatest repellent effect, as shown by the lowest RC50 dose, followed by O. minutiflorum and O. majorana. Interestingly, this means that the oil that was least effective in killing the larvae was the most effective in repelling them. The calculated synergistic factor of any combination was higher than 1 which means that combinations have a synergistic effect. In conclusion, the combination of all three oils showed higher toxic and repellent activities than either oil separately or combinations of any two oils, suggesting synergistic effects with low doses. Further studies including field trials and the establishment of the mode of action and side effects are urgently needed to expand on these findings, and other tick stages such as adults should also be tested.

Authentication of the Botanical and Geographical Origin and Detection of Adulteration of Olive Oil Using Gas Chromatography, Infrared and Raman Spectroscopy Techniques: A Review.

Olive oil is among the most popular supplements of the Mediterranean diet due to its high nutritional value. However, at the same time, because of economical purposes, it is also one of the products most subjected to adulteration. As a result, authenticity is an important issue of concern among authorities. Many analytical techniques, able to detect adulteration of olive oil, to identify its geographical and botanical origin and consequently guarantee its quality and authenticity, have been developed. This review paper discusses the use of infrared and Raman spectroscopy as candidate tools to examine the authenticity of olive oils. It also considers the volatile fraction as a marker to distinguish between different varieties and adulterated olive oils, using SPME combined with gas chromatography technique.

The Use of Essential Oil and Hydrosol Extracted from Satureja hellenica for the Control of Meloidogyne incognita and M. javanica.

Essential oil (EO) and hydrosol (HL) isolated from an indigenous plant species Satureja hellenica were evaluated against Meloidogyne incognita and M. javanica. Particularly, the activity of extracts on a second stage juvenile's (J2s) motility, the hatching of J2s from eggs, egg differentiation and the effect on J2s in soil were tested. A paralysis of 100% of the J2s of both species was recorded after 96 h of immersion in the essential oil, at a dose of 2000 µL/L. At the same dose, the percentage of paralyzed J2s after 48 h of immersion was more than 80%, for both Meloidogyne species. The use of hydrosol has shown encouraging results only in the dilution of 50%, where for both Meloidogyne species tested, the percentage of paralyzed J2s was more than 70% after 48 h of immersion, while the percentage was increased to 90% after 96 h of immersion. Egg differentiation was ceased after immersion, either in EO or HL. However, this decrease in egg differentiation was evident only at higher concentrations of EO and at the highest HL dilution (0.5 v/v). The hatching of M. incognita J2s was decreasing as the dose was increasing. The lowest numbers of hatched J2s were recorded at the doses of 2000 and 4000 µL/L. A clear reduction in M. javanica J2s hatching was observed as the dose was increased to 250 µL/L, a fact constantly observed as the dose was increasing up to 4000 µL/L. Lower numbers of nematodes were recorded in roots grown in infested soil after the application of EO or HL at the highest doses. The EO of S. hellenica is characterized by the presence of p-cymene (27.46%) and carvacrol (23.25%), and in a lesser extent of other constituents, such as borneol (6.79%), carvacrol methylether (6.77%), γ-terpinene (4.63%) and 4-terpineol (3.65%). Carvacrol was the major constituent found in the HL (50.12%), followed by borneol and 4-terpineol (20.42 and 6.72%, respectively).

Evaluation of Eight Essential Oils for Postharvest Control of Aspergillus carbonarius in Grapes.

ABSTRACT: A range of fungal species are associated with postharvest spoilage of grapes. However, Aspergillus carbonarius is the primary fungus responsible for the contamination of grapes with ochratoxin A, a mycotoxin causing several confirmed negative health effects in humans and animals. Aiming to find a method, safe for consumers, to prevent postharvest decay and ochratoxin A contamination of grapes, the potential use of essential oils as preservatives was investigated. Essential oils of Origanum dictamnus (dittany), Origanum onites (oregano), Origanum microphyllum (marjoram), Thymbra capitata (thyme), Satureja thymbra (savory), Rosmarinus officinalis (rosemary), Laurus nobilis (laurel), and Salvia officinalis (sage) were tested. The essential oil components were identified by gas chromatography-mass spectrometry analysis. A first evaluation of the effectiveness of essential oils was performed in vitro at a range of concentrations up to 300 µL L-1. Based on the results of the in vitro tests, the four most effective essential oils (O. dictamnus, O. onites, T. capitata, and S. thymbra) were tested on Sultana grapes during postharvest storage. The four essential oils tested, which had carvacrol and/or thymol as a common component, at a high concentration significantly reduced or even inhibited growth of the fungus in all treatments. As revealed from the results, the essential oils of O. dictamnus, O. onites, and S. thymbra were the most effective, causing total inhibition of the growth of the fungus with a minimum concentration of 100 µL L-1, followed by the essential oil of T. capitata, which showed total effectiveness with a minimum concentration of 200 µL L-1. Although essential oils of O. microphyllum, L. nobilis, S. officinalis, and R. officinalis had a significant effect on the growth of A. carbonarius, they failed to inhibit its growth at any of the concentrations tested.

Encapsulation of Mentha pulegium Essential Oil in Yeast Cell Microcarriers: An Approach to Environmentally Friendly Pesticides.

A green approach for the encapsulation of Mentha pulegium essential oil in commercial baker's yeast and its evaluation as a pesticide against the insect pest Myzus persicae are presented. Upon treating aqueous yeast cell dispersion with the essential oil, the formation of essential-oil-loaded microparticles of about 9 µm is observed, with a loading capacity ranging from 29 to 36%, depending upon the encapsulation conditions. The thermal properties of the microparticles were characterized using differential scanning calorimetry and thermogravimetric analysis, confirming the protection of the essential oil from the cells. Encapsulation prolonged the insecticidal activity of the essential oil by 3 days.

Direct determination of rosmarinic acid in Lamiaceae herbs using diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) and chemometrics.

For the determination of rosmarinic acid (RA) directly in pulverized plant material, a method is developed using diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) without any physicochemical pretreatment of samples. The RA content of 11 samples of eight different Lamiaceae herbs, as determined by high performance liquid chromatography (HPLC), varied between 86 ± 1 mg/g (in lemon balm) and 12.0 ± 0.8 mg/g (in hyssop) of dried plant material. The 11 samples and 50 other additional samples, which were prepared by mixing initial samples with KBr, were measured using DRIFTS. The second derivative of the spectral region 1344-806 cm(-1) was used and the corresponding data were analyzed using partial least squares (PLS) regression. The correlation between infrared spectral analysis and HPLC measurements shows that the DRIFTS method is sufficiently accurate, simple, and rapid. The RA content of the 11 Lamiaceae samples determined by DRIFTS ranged from 81 ± 4 mg/g (in lemon balm) to 12 ± 3 mg/g (in hyssop) of dried plant material.

Phenolic acid contents, essential oil compositions and antioxidant activities of two varieties of Salvia euphratica from Turkey.

This study was designed to evaluate the in vitro antioxidant activities of n-hexane (Hex), dichloromethane (DCM), methanol (MeOH) and essential oils (EO) extracts obtained from Salvia euphratica var. euphratica and Salvia euphratica var. leiocalycina and to determine their essential oil and phenolic acid compositions. The samples were screened for their antioxidant activity by using DPPH and ß-carotene/linoleic acid assays. Methanol extracts of both varieties exhibited strong antioxidant activities. Our results showed that rosmarinic acid was dominant phenolic acid of MeOH extracts (39.4 and 55.8 µg mg⁻¹, respectively). The chemical compositions of essential oils of two varieties were analysed and their main components were determined as eucalyptol (18.4%) and trans-pinocarvyl acetate (24.9%), respectively. It can be said that these varieties could be used as natural antioxidant.

Studies on the antioxidant activity of the essential oil and methanol extract of Marrubium globosum subsp. globosum (lamiaceae) by three different chemical assays.

This study is designed to examine the chemical composition and in vitro antioxidant activity of the essential oil and sub-fractions of the methanol extract of Marrubium globosum subsp. globosum. The GC and GC-MS analysis of the essential oil were resulted in the determination of 84 components representing 88.2% of the oil. The major constituents of the oil were spathulenol (15.8%), beta-caryophyllene (9.0%), caryophyllene oxide (7.9%), germacrene D (6.5%), and bicyclogermacrene (3.1%). Antioxidant activities of the samples were determined by three different test systems namely DPPH, beta-carotene/linoleic acid and reducing power assay. In DPPH system, the weakest radical scavenging activity was exhibited by the essential oil (1203.38+/-7.18 microg ml(-1)). Antioxidant activity of the polar sub-fraction of methanol extract was superior to the all samples tested with an EC(50) value of 157.26+/-1.12 microg ml(-1). In the second case, the inhibition capacity (%) of the polar sub-fraction of methanol extract (97.39%+/-0.84) was found the strongest one, which is almost equal to the inhibition capacity of positive control BHT (97.44%+/-0.74). In the case of reducing power assay, a similar activity pattern was observed as given in the first two systems. Polar sub-fraction was the strongest radical reducer when compared with the non-polar one, with an EC(50) value of 625.63+/-1.02 microg ml(-1). The amount of the total phenolics was highest in polar sub-fraction (25.60+/-0.74 microg/mg). A positive correlation was observed between the antioxidant activity potential and total phenolic level of the extracts. On the other hand, total flavonoid content was found equal for the both sub-fractions.

Comparison of distillation and ultrasound-assisted extraction methods for the isolation of sensitive aroma compounds from garlic (Allium sativum).

A comparative study of traditional simultaneous distillation extraction (SDE), microwave assisted hydrodistillation extraction (MWHD) and ultrasound-assisted extraction (USE) is presented, for the extraction of essential oils from fresh garlic (Allium sativum) cloves. Each method is evaluated in terms of qualitative and quantitative composition of the isolated essential oil. The highly reactive sulfur molecules of the garlic volatile fraction show variable response to the different isolation methods. The application of ultrasound for the extraction of the essential oil is considered to cause a lesser damage of thermal-sensitive molecules, thus, providing a better approach of the compounds primarily responsible for the characteristic odor and taste of freshly chopped garlic. All heat-involving isolation procedures have been shown to differentiate the volatile-fraction profile as analyzed by GC-MS. Especially when grouping the compounds into cyclic and acyclic, the percentage concentrations drop from 77.4% to 8.7% for the acyclic while that of the cyclic compounds increase from 4.7% to 70.8%. The observed fact may be attributed to the effect of the heat applied, which changes from harsh thermal treatment (SDE) to short time thermal (MWHD) and room-temperature isolation (USE). The use of USE proves to be crucial in order to provide reliable insight into garlic's chemistry.

Rapid method for simultaneous quantitative determination of four major essential oil components from oregano (Oreganumsp.) and thyme (Thymus sp.) using FT-Raman spectroscopy.

A simple and rapid method for the quantitative determination of four major components found in oregano and thyme essential oils is presented. The method correlates the Raman peak intensity in the spectral region from 1800 to 600 cm(-1) and the concentration percentage of each particular constituent in the sample. To achieve accurate quantification results and avoid the risk of overlapping peaks of unknown Raman-active substances in natural essential oils, the peaks must be analyzed. For this purpose, PEAKSOLVE software (Ver. 1.0.5) was used. Unknown samples were measured with the FT-Raman method, and the results were compared to those of the gas chromatographic (GC) analysis. The comparison was made at a confidence level of 99%, and the two methods scored equally in terms of repeatability and accuracy even at the edge of the method specifications. The new method can provide accurate results in very short times once the setup is complete and could be utilized in areas where vast amounts of samples must be analyzed.

The in vitro antioxidant and antimicrobial activities of the essential oil and methanol extracts of Achillea biebersteini Afan. (Asteraceae).

The essential oil and methanol extracts from A. biebersteinii Afan. (Asteraceae) were evaluated for their antimicrobial and antioxidant activities in vitro. The oil showed stronger antimicrobial activity than the extracts. Their antioxidant features were also evaluated using diphenyl-1-picrylhydrazyl (DPPH), inhibition of superoxide and hydroxyl radicals and inhibition of the lipid peroxidation assays. Particularly, polar subfraction of the methanol extract showed antioxidant activity. The GC-MS analysis of the oil has resulted in the identification of 23 components; piperitone, eucalyptol, camphor, chrysanthenone and borneol were the main components. Antimicrobial activity tests carried out with the fractions of the oil showed that the activity was mainly observed in those containing eucalyptol and camphor, in particular, followed by borneol and piperitone.

Qualitative determination of volatile compounds and quantitative evaluation of safranal and 4-hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde (HTCC) in Greek saffron.

Safranal (2,6,6-trimethyl-1,3-cyclohexadiene-1-carboxaldehyde) is the main component of saffron's essential oil. It was obtained using microsimultaneous hydro distillation-extraction (MSDE) and by ultrasound-assisted extraction (USE), which is a mild method. 4-Hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde (HTCC) is a precursor of safranal and was obtained in considerable amounts only by USE. Five C(13)-norisoprenoids were found in saffron for the first time. Using a gas chromatography technique, safranal and HTCC were quantified from Greek saffron samples. The quantity of safranal isolated by MSDE ranged between 288.1 and 687.9 mg/100 g of saffron, whereas in the case of USE safranal and HTCC ranged between 40.7 and 647.7 mg/100 g of saffron and between 41.7 and 397.7 mg/100 g of saffron, respectively. Freeze-drying was also tested as an alternative drying method. Over years of storage at 4 degrees C the quantity of safranal remained mostly constant while the quantity of HTCC decreased over the same periods.

In vitro antioxidant, antimicrobial, and antiviral activities of the essential oil and various extracts from herbal parts and callus cultures of Origanum acutidens.

The essential oil and various extracts obtained from Origanum acutidens and methanol extracts (MeOH) from callus cultures have been evaluated for their antioxidative, antimicrobial, and antiviral properties. The essential oil exhibited strong antimicrobial activity with a significant inhibitory effect against 27 (77%) of the 35 bacteria, 12 (67%) of the 18 fungi, and a yeast tested and moderate antioxidative capacity in DPPH and beta-carotene/linoleic acid assays. GC and GC-MS analyses of the oil resulted in the identification of 38 constituents, carvacrol being the main component. The MeOH extracts obtained from herbal parts showed better antioxidative effect than that of butylated hydroxytoluene (BHT), whereas callus cultures also exhibited interesting antioxidative patterns. Concerning antiviral activity, none of the extracts inhibited the reproduction of influenza A/Aichi virus in MDCK cells. The MeOH extracts from herbal parts inhibited the reproduction of HSV-1, and also callus cultures exerted slight antiherpetic effect.

In vitro antimicrobial and antioxidant activities of the essential oils and various extracts of Thymus eigii M. Zohary et P.H. Davis.

This study was designed to examine the in vitro antimicrobial and antioxidant activities of the essential oil and various extracts obtained from aerial parts of Thymus eigii. The essential oil was particularly found to possess stronger antimicrobial activity, whereas other nonpolar extracts and subfractions showed moderate activity and polar extracts remained almost inactive. GC-MS analysis of the oil resulted in the identification of 39 compounds, representing 93.7% of the oil; thymol (30.6%), carvacrol (26.1%), and p-cymene (13.0%) were the main components. The samples were also subjected to a screening for their possible antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and beta-carotene-linoleic acid assays. In the former case, the polar subfraction of the methanol extract was found to be superior to all extracts tested, only 16.8 microg/mL of which provided 50% inhibition, whereas all extracts, particularly the polar ones, seem to inhibit the oxidation of linoleic acid in the latter case. These data were further supported by total phenolics analysis, indicating that the antioxidative potential of the extracts was closely related to their phenolic constituents.

Antimicrobial activity of essential oil and methanol extracts of Achillea sintenisii Hub. Mor. (Asteraceae).

The essential oil, obtained by Clevenger distillation, and water-soluble and water-insoluble parts of the methanol extracts of Achillea sintenisii Hub. Mor. were individually assayed for their antimicrobial activities against 12 bacteria and two yeasts, Candida albicans and C. krusei. No activity was exhibited by the water-soluble subfraction, whereas both the water-insoluble subfraction of the methanol extracts and the essential oil were found to be active against some test microorganisms studied. Since the essential oil possessed stronger activity than the other extracts tested, it was further fractionated and the fractions were evaluated for their antimicrobial activity, followed by GC-MS analysis, resulting in the identification of 32 compounds which constituted 90.2% of the total oil. The GC-MS analysis of the oil and its fractions revealed that the main components of the oil, e.g. camphor and eucalyptol, possessed appreciable activity against C. albicans and Clostridium perfringens. The fi ndings presented here also suggest that the other constituents of the oil, e.g. borneol and piperitone can also be taken into account for the activity observed.

Antioxidant and antimicrobial activity of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae).

The in vitro antimicrobial and antioxidant activities of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae) were investigated. GC-MS analysis of the essential oil resulted in the identification of 36 compounds constituting 90.8% of the total oil. Eucalyptol, camphor, alpha-terpineol, beta-pinene, and borneol were the principal components comprising 60.7% of the oil. The oil strongly reduced the diphenylpicrylhydrazyl radical (IC(50)=1.56 micro g/ml) and exhibited hydroxyl radical scavenging effect in the Fe(3+)-EDTA-H(2)O(2) deoxyribose system (IC(50)=2.7 micro g/ml). It also inhibited the nonenzymatic lipid peroxidation of rat liver homogenate (IC(50)=13.5 micro g/ml). The polar phase of the extract showed antioxidant activity. The oil showed antimicrobial activity against Streptococcus pneumoniae, Clostridium perfringens, Candida albicans, Mycobacterium smegmatis, Acinetobacter lwoffii and Candida krusei while water-insoluble parts of the methanolic extracts exhibited slight or no activity. This study confirms that the essential oil of Achillea millefolium possesses antioxidant and antimicrobial properties in vitro.