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In this paper, a detailed study on chemical characterization of essential oils (EOs) constituents of leaves and stems of Achillea fragrantissima were carried out using GC-FID and GC–MS analysis employing two different stationary phase columns. In the studied plant which is collected from the central region of Saudi Arabia, trans-sabinyl acetate and trans-sabinol have been identified as the major components. To the best of our knowledge, these components are being reported for the first time as the major constituents in the EOs of A. fragrantissima. The results showed that chief chemical components of both (leaves and stems) oils were found to be almost same, however, their contents varied noticeably from each other. Among 108 identified components in the leaves oil, the major components were trans-sabinyl acetate (20.7 ± 0.00), trans-sabinol (14.9 ± 0.13), artemisia ketone (12.7 ± 0.46), santolina alcohol (10.1 ± 1.30), β-sesquiphellandrene (5.5 ± 0.01), β-thujone (5.1 ± 0.11). Whereas, in the stem oil 85 components were identified and trans-sabinyl acetate (24.0 ± 0.19), trans-sabinol (19.2 ± 0.01), artemisia ketone (16.3 ± 0.74), santolina alcohol (10.4 ± 1.50), and β-sesquiphellandrene (4.8 ± 0.01) were found to be the major components. Among the identified components form both oils, 23 components were specific to only leaves oil, whereas 85 components were found to be common in both oils.

The plant produce metabolites can be improved by biotechnology methods, one of the most common techniques is to establish a regeneration system through plant tissue culture. Santolina chamaecypatissus L. is a valuable aromatic species used in pharmaceutical industry and ornamental field. Inefficient propagations methods limit agricultural utilization of S. chammaecypatissus to produce volatile chemical constituents. Here, we reported a rapid and reliable in vitro propagation protocol for S. chamaecypatissus that showed higher accumulation of volatile compounds. In vitro cultures were obtained by an optimized sterilization protocol using 75% ethanol combined with 1% sodium hypochlorite. Callus induction from stems was highest with 1 mg/L benzyladenine (BA) and 0.2 mg/L naphthaleneacetic acid (NAA). The shoot regeneration was improved by adjusting plant growth regulators to an optimal concentration of 0.04 mg/L BA and 0.1 mg/L NAA followed by root induction with 0.1 mg/L NAA. Obtained micropropagated plants were morphologically stable and grew vigorously with a 100% survival rate in soil matrix consisting of sand, gravel, and perlite. Evaluation of the chemical composition of volatiles via gas chromatography-mass spectrometry (GC–MS) of micropropagated plants on field to regular field plants revealed that the volatile composition accumulated in micropropagated plants higher than regular field plants. The rapid and efficient micropropagation protocol established in this study strongly contributes to overcome limitations of cultivation of S. chamaecypatissus, a valuable feedstock for extraction of bioactive phytochemicals.

Ex situ conservation of Bulgarian endemic plant Achillea thracica Velen. was achieved by successful in vitro cultivation of mono-nodal segments on MS-B5 medium supplemented with 1.0 mg/L BA for 20 days and subsequent transferring of regenerated plants on hormone free basal MS-B5 medium for root development and accumulation of leaf biomass. In vitro multiplicated plants were successfully acclimated in a growth chamber with 100% survival. GC–MS analysis of the essential oils resulted in the identification of 30, 10 and 28 compounds in in situ grown, in vitro cultivated and ex vitro adapted plants, respectively, constituting 77.7%, 99.9% and 84.1% of the total oils. The wider variety of compounds was found in the essential oils of in situ and ex vitro adapted plants where santolina alcohol, β-eudesmol, 1,8-cineole, germacrene D, α-cadinol and artemisia alcohol were the principal components comprising 68.7% and 69.3 of the oil, respectively. In vitro cultivated plants consist of mainly 1,8-cineole, germacrene D and artemisia alcohol representing 87% of the oil. Different growth conditions affect the composition of essential oils, suggesting their possible involvement in the process of adaptation and surviving in changing environmental conditions.

The antimicrobial properties of essential oil from various Santolina species have not been investigated enough in the previous studies dealing with the biological activities of medicinal plants. In Tunisia, Santolina chamaecyparissus L. (Asteraceae) is the only Santolina species recorded and is used as vermifuge and emmenagogue. The chemical composition, antibacterial and antifungal properties of essential oils from the flowerheads and roots of spontaneous S. chamaecyparissus growing in Tunisia and the chemical composition which leads to the Tunisian chemotype are investigated here for the first time. Essential oils isolated by hydro distillation from flowerheads and roots of S. chamaecyparissus were analyzed by GC and GC/MS. Two methods served for antimicrobial assays of the essential oils: diffusion in a solid medium and micro-well dilution assay. Antifungal tests were carried out by the agar incorporation method. Sixty-seven constituents were identified from the essential oil of the flowerhead. The major constituents were: 1,8-cineole and β-eudesmol. Two non identified compounds were present at the highest concentration in root oil. Flowerhead oil was characterized by high contents in monoterpenes and sesquiterpenes oxygenated compounds. The flowerhead essential oil demonstrated potent of antibacterial properties against Pseudomonas aeruginosa ATCC and Enterococcus faecalis ATCC, with MIC of 0.625μg/ml. These findings demonstrate that the flowerhead essential oils of S. chamaecyparissus have excellent antibacterial properties and for this reason they could contribute to decrease the problem of microbial resistance to antibiotics.

Cumin seeds were collected from agro-ecological sub-regions comprising of major cumin growing areas of Rajasthan and Gujarat state, India, to analyze chemical composition of essential oil. The study area was located between the geographical co-ordinates 21°78′ 32″N, 71°33′85″E to 26°21′ 28″ N, 74°37′61″ E. Maximum and minimum EO yield (35.9–28.4%) were obtained in cumin from AESR 2.4 and 5.1 respectively. GC–MS analysis of EO indicated the preponderance of terpenic hydrocarbons in the AESR 2.4 (53.85%) and decrease in AESR 2.1 (32.62%) while cuminaldehyde content was more (39.90%) in AESR 4.2 and less (25.84%) in 2.4. The prominent terpenic compounds included γ-terpinene, β-pinene, santolina triene, α-terpinene, α-pinene and β-thujene, the concentration of these compounds varied with AESRs. Cumic alcohol (10.33–14.40%), anethol & estragol (4.03–9.15%) were the major alcohols with cymol (2.9–4.26%) and geraniol (0.02–2.39%) in traces. Total phenolic content in the various AESRs ranged from 41.50 (AESR 5.1) to 58.61 mg GAEg −1 seeds (AESR 2.3) while the flavonoid content in the AESRs ranged between 27.14–36.03 mg QE g −1 seeds, being least in 4.2 and highest in 5.1. The study revealed that the average radical scavenging percentage of methanol seed extract does not vary with the different AESRs (94.98–99.05%), as well as EC 50 value is also same (15.31–15.38 mg BHTE) for samples from different AESRs. Cumin from the AESRs with favourable minimum and maximum temperature and rainfall showed high cuminaldehyde bearing pleasant aroma whereas the AESRs with biotic/abiotic stresses showed higher terpenic compounds with increased pungency.

The inhibitive action of Santolina chamaecyparissus extract as a natural inhibitor on corrosion of 304 stainless steel in 3.5% NaCl solution was investigated by potentiodynamic polarization, EIS, and SEM techniques. The results indicate that the extract shows mixed-type inhibition behavior and inhibition efficiency of 86.9% was achieved with the addition of 1.0 g L−1 extract. Impedance curves show that increasing Santolina concentration increases charge transfer resistance and decreases double layer capacitance. The corrosion rate decreased with increasing concentration of extract and increased with the increase in temperature of the medium. Adsorption of the extract obeyed the Langmuir isotherm model.

Ethnopharmacological relevance Santolina species are widely used in traditional medicine in the Mediterranean region for their anti-inflammatory, antimicrobial, antispasmodic, digestive, and analgesic properties. S. impressa, a Portuguese endemism, is traditionally recognized for its beneficial anti-inflammatory properties in several gastrointestinal affections and is also used in oropharyngeal infections. Aim of the study The present study aims to characterize the essential oil of S. impressa growing in Portugal and validate its traditional uses by assessing the anti-inflammatory potential of its essential oil at concentrations without toxicity. The antifungal properties of the oil are also addressed, as well as, the putative mechanism of action underlying these effects. Material and methods The essential oil was obtained in accordance with the European Pharmacopoeia and characterized by GC and GC-MS. The anti-inflammatory potential of the oil was assessed on LPS-stimulated macrophages, through the production of nitric oxide (NO) using the Griess reaction. Putative mechanisms of action included the role of the oil as a NO scavenger, as well as its effect on the expression of two key pro-inflammatory enzymes, iNOS and COX-2 by Western blot analysis. The antifungal effect of the oil was evaluated according to the CLSI guidelines on several yeast and filamentous strains and on two major virulence factors in Candida albicans, namely germ tubes and biofilms. Ultrastructural modifications on dermatophytes were also unveiled by transmission electron microscopy. Results S. impressa essential oil was primarily characterized by the presence of monoterpene hydrocarbons and oxygenated monoterpenes, being the main compounds β-pinene (22.5%), 1,8-cineole (10.0%), limonene (9.1%), camphor (8.1%) and β-phellandrene (8.0%). A significant decrease (ca 60.0%) in nitrite levels was observed in LPS-stimulated macrophages treated with the oil without affecting cell viability. This effect could be explained by a great reduction on iNOS expression (85.0% inhibition), thus underpinning the anti-inflammatory potential of the oil. The oil also showed a fungicidal effect, being more active against Cryptococcus neoformans, Epidermophyton floccosum and Trichophytum rubrum. For these dermatophytes, significant ultrastructural modifications in cell wall structure were detected. Strikingly, for C. albicans, the oil showed a significant anti-infective potential (at 0.07 mg/mL for germ tube inhibition and 0.02 mg/mL for biofilm disruption) before fungal growth inhibition occurred. Conclusions Our results validate the main traditional use ascribed to S. impressa, namely its anti-inflammatory effect. In addition, an antifungal potential is pointed out, thus corroborating the antimicrobial uses and adding new value to an endemic species poorly recognized by the industry.

Essential oils of Achillea biebersteinii , A. santolina and A. mellifolium were obtained by hydrodistillation, then analyzed using gas chromatography (GC) and GC/mass spectrometry (MS). The obtained oils showed considerable toxic and growth inhibitory activities against the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). The time of exposure, the technique used and the stage treated, all were activity-determinant factors, where A. biebersteinii oil showed the strongest insecticidal activity and the adult stage was more susceptible than larvae. Using topical application, LC 50 values of A. biebersteinii , A. santolina and A. mellifolium oils after 96 h of exposure of larvae were 30.3, 47.8 and 62.3 μg/mg insect, respectively. In this case, LC 50s against adults were ranged between 21.8 and 50.7 μg/mg insects. Using treated filter papers, the LC 50 values after 96 h of exposure against larvae were ranged between 124.9 and 212.0 μL/cm 2 , while against adults were ranged between 113.0 and 196.8 μL/cm 2 . In the fumigant bioassay, LC 50s after 96 h of exposure against larvae were between 41.7 and 93.2 μL/L air, while against adults were between 36.1 and 80.6 μL/L air. When prepared as nano-emulsions and tested as fumigants, toxicity of oils was increased dramatically, where LC 50 values after 96 h of exposure against larvae and adults were ranged between 11.0 and 27.5 μL/L air and 8.8 and 21.3 μL/L air, respectively. When exposed as 2nd instar larvae to sub-lethal doses-treated grains, the developmental course, life span and F 1 progeny of T. castaneum were significantly affected, where the oil of A. santolina showed the strongest activity. All of these developmental disruptions led to a great reduction in the number of adults that undergo successful emergence.

Amplified fragment length polymorphism (AFLP) markers were used to assess the genetic diversity of 57 Achillea accessions belonging to five species, A. millefolium , A. filipendulina , A. tenuifolia , A. santolina and A. biebersteinii . Nine AFLP primer combinations were used, which produced 301 polymorphic bands. In most species, a high level of genetic variation was detected among the genotypes. The Jaccard's similarity indices (J), based on AFLP profiles, were subjected to UPGMA cluster analysis. Application of Mantel's test for cophenetic correlation to the cluster analysis indicated the high fitness of the accessions to a group ( r = 0.918). The dendrogram generated revealed five major groups corresponding to five species. The principle coordinate analysis (PCoA) data confirmed the results of the clustering. Among the species, A . teunifolia and A. santolina showed the greatest and the least genetic diversity, respectively. A . filipendulina accessions were acquired primarily from the same ecological regions of western Iran. Accessions belonging to A . biebersteinii originated from the Isfahan province and were separated from other species at the root of the dendrogram. The results of the clustering method, based on AFLP markers, corresponded closely with the geographical origins of the genotypes. The results of the present study could contribute to a better understanding and management of conservation and exploitation of the Achillea germplasm.

The compositions of essential oils of 19 accessions belonging to six different Achillea species, transferred from the natural habitats in 10 provinces of Iran to the field conditions, were assessed. The relationship between the leaf areas of selected accessions with their essential oil content was also investigated. Essential oil yield of dried plants obtained by hydro-distillation ranged from 0.1 to 2.7% in leaves. Results indicated a significant variation in oil composition among and within species. Total of 94 compounds were identified in 19 accessions belonging to the six species of A. millefolium , A. filipendulina , A. tenuifolia , A. santolina , A. biebersteinii and A. eriophora . The major constituents of the leaves in the tested genotypes were determined as germacrene-D, bicyclogermacrene, camphor, borneol, 1,8-cineole, spathulenol and bornyl acetate. According to the major compounds, four chemotypes were defined as: (I) spathulenol (1.64–34.31%) + camphor (0.2–15.61%) (7 accessions); (II 1 ) germacrene-D (18.78–23.93%) + borneol (7.93–8.26%) + bornyl acetate (11.56–14.66%) (5 accessions); (II 2 ) germacrene-D (13.28–36.28%) + bicyclogermacrene (5.93–8.4%) + 1,8-cineole (15.26–19.41%) + camphor (14.95–23.32%) (2 accessions); (III) borneol + camphor (52.04–63.27) (2 accessions); (IV) germacrene-D (45.86–69.64%) (3 accessions). The relationships of chemotypes with soil type and climatic conditions of collected regions were assessed, as probable reasons of high variations in essential oil components, and discussed.

International audience; A detailed analysis of Santolina corsica essential oil was carried out by combination of GC (RI), GC-MS and 13C NMR spectroscopy. After fractionation by column chromatography, 50 components were identified, accounting for 88.2% of the total amount of the oil. The chemical composition was dominated by monoterpene hydrocarbons, myrcene (34.6%), santolina triene (13.5%) and beta-phellandrene (11.7%). Beside the main compounds, we noted the occurrence of irregular mono and sesquiterpenes belonging to five families: santolinane, artemisane, chrysanthemane, lavandulane and sesquilavandulane. Three compounds, lyratyl butyrate, isolyratone and epi-isolyratol were isolated and their structure elucidated by 2D NMR. Antibacterial activity was tested against six bacteria strains. The essential oil was effective against Staphylococcus aureus and C. jejuni. Lyratol was identified as main responsive of the antibacterial activity. The content of lyratol was measured in 33 oil samples isolated from individual plants.

Oxidative stress is produced under diabetic condition and is likely involved in progression of pancreatic damage found in diabetes. In the present study, we examined possible protective effect of Achillea santolina L. (Compositae) against pancreatic damage in streptozotocin (STZ)-treated diabetic rats. Achillea santolina extract (ASE) is used by the traditional healers in many part of Iraq, as a hypoglycaemic agent. We evaluated the effect of ASE on blood glucose level, serum nitric oxide (NO) concentration and the oxidative stress status in rat pancreatic tissue. STZ was injected intraperitonealy at a single dose of 40 mg kg −1 to induce diabetes. ASE (0.1 g/kg day) was orally administered to a group of diabetic rats for 30 consecutive days. Results showed significant reduction in the activities of superoxide dismutase (SOD), catalase (CAT) and pancreatic glutathione (GSH) levels in the diabetic rats compared to the control subjects. On the other hand, blood glucose level, serum NO, malondialdehyde (MDA), a marker of lipid peroxidation, protein oxidation indices including protein carbonyl (PCO) and advanced oxidation protein products (AOPP) were significantly elevated in pancreas of the diabetic group. Treatment with ASE reduced blood glucose level, serum NO, pancreatic MDA, PCO and AOPP. In addition, the content of GSH was restored to the normal level of the control group. Furthermore, ASE significantly increased CAT and SOD activities in ASE-treated rats. Based on our data, it can be concluded that Achillea santolina have a high hypoglycaemic activity and this may be attributed to its antioxidative potential. © 2007 Elsevier Ireland Ltd. All rights reserved.

The antioxidative activities of hydroalcoholic extract of Achillea santolina were investigated employing various established in vitro systems including total antioxidant activity in linoleic acid emulsion system, 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals scavenging, reducing power, and inhibitory effect on protein oxidation as well as the inhibition of Fe 2+ /ascorbate induced lipid peroxidation in rat liver homogenate. Total phenolic and flavonoid content of A. santolina extract (ASE) was also determined by a colorimetric method. The results revealed that ASE has notable inhibitory activity on peroxides formation in linoleic acid emulsion system along with concentration-dependent quenching of DPPH and superoxide radicals. Furthermore, the extract showed both nonsite-specific (Fe 2+ + H 2 O 2 + EDTA) and site-specific (Fe 2+ + H 2 O 2 ) hydroxyl radical scavenging suggesting potent hydroxyl radical scavenging and chelating ability for iron ions in deoxyribose degradation model. A linear correlation between ASE and the reducing power was also observed ( r 2 = 0.9981). ASE prevents thiobarbituric acid reactive substances formation in Fe 2+ /ascorbate induced lipid peroxidation in rat liver tissue in a dose-dependent manner. Moreover, free radical induced protein oxidation was suppressed significantly by the addition of ASE over a range of concentration. These results clearly demonstrated that A. santolina extract possess a marked antioxidant activity.

A search for naturally occurring drugs with antifungal activity lead to Santolina oil, a volatile oil distillate of Santolina chamaecyparissus. The studies revealed that Santolina oil was effective in controlling experimental candidiasis in vitro and in vivo. It had a synergistic effect on clotrimazole in controlling Candida albicans in vitro. It significantly controlled experimental vaginal candidiasis and experimental systemic candidosis. Santolina oil was able to control the superficial cutaneous mycoses. It is recommended as a potential candidate for further studies, including clinical studies.

Five plants (Myrtus communis, Apium graveolens, Matricaria chamomilla, Withania somnifera and Achillea santolina) grown in Iraq were assessed for their anti-inflammatory activity on intact rats by measuring the suppression of carrageenan-induced paw edema produced by 1/10 of the intraperitoneal LD50 doses for the respective 80% ethanol extracts. Acetylsalicylic acid was used as the standard drug. Results showed that the plants possessed varying degrees of anti-inflammatory activity and were classified in the following descending order of activity: W. somnifera greater than A. graveolens greater than A. santolina greater than M. chamomilla greater than M. communis.

The volatile oil of Artemisia arbuscula arbuscula contained a new irregular monoterpene, 2,5-dimethyl-4-vinyl-1,5-hexadiene-3-ol (isolyratol), which was isolated and identified by spectral means. The optically pure furanoid (2S,5S)-trans-5-methyl-5-vinyltetrahydrofur-2-yl methyl ketone (arbusculone), was also characterized by transformation to known (2S,5S)-trans-linalyl oxide. The former component has never been isolated from natural sources prior to this study. The neutral pentane extract also contained several previously characterized non-head-to-tail monoterpenes including artemiseole, artemisia ketone, artemisyl acetate, methyl santolinate, and santolina triene, as well as the regular monoterpenes 1,8-cineole, camphor, p-cymene, camphene and the C6 fragment, terelactone.

The antibacterial activity of Artemisia herba-alba was investigated. Only its essential oil was active against some Gram-positive and Gram-negative bacteria. The essential oil was fractionated by column chromatography, and these fractions were tested for antibacterial activity. The principal component of the most active fraction was santolina alcohol.