Stachys Species Research Articles

Valorisation of stachysetin from cultivated Stachys iva Griseb. as anti-diabetic agent: a multi-spectroscopic and molecular docking approach

Stachys species are considered as important medicinal plants with numerous health benefit effects. In continuation of our research on the Greek Stachys species, the chemical profile of the aerial parts of cultivated S. iva Griseb. has been explored. The NMR profiles of the plant extract/infusion were used to guide the isolation process, leading to the targeted isolation of seventeen known compounds. The rare acylated flavonoid, stachysetin, was isolated for the third time from plant species in the international literature. Identification of the characteristic signals of stachysetin in the 1D 1H-NMR spectrum of the crude extract was presented. In order to evaluate the potential of the identified chemical space in Stachys to bear possible bioactivity against diabetes, we performed an in silico screening against 17 proteins implicated in diabetes, as also ligand based similarity metrics against established anti-diabetic drugs. The results capitalized the anti-diabetic potency of stachysetin. Its binding profile to the major drug carrier plasma protein serum albumin was also explored along with its photophysical properties suggesting that stachysetin could be recognized and delivered in plasma through serum albumin and also could be tracked through near-infrared imaging. Communicated by Ramaswamy H. Sarma

Chemical Constituents of the Essential Oil of Stachys fruticulosa M. Bieb. From Iran

Background: Stachys (Lamiaceae) is a genus with varied effects and applications in traditional medicine. The essential oil (EO) of different species of genus Stachys has received much attention in different studies. According to previous evidence, the components of EO in different species have diversity although there is no previous data regarding investigating Stachys fruticulosa for EO. Considering the importance of the plants of genus Stachys, vast types of compounds in the EO of the genus, the aim of the present study was to evaluate the components of EO of S. fruticulosa M. Bieb. Methods: The EO of the aerial parts from S. fruticulosa was obtained by hydrodistillation and then the oil was analyzed by gas chromatography/mass spectrometry (GC/MS) and 95% of the oil (17 components) was identified accordingly. The identity of the components was assigned by comparing their mass spectra and retention indices with those of authentic samples. Results: Most oil components were α-terpinyl acetate (24.6%), 3-n-Butyl phthalide (20.5%), p-cymene (18.2%), and β-phellandrene (18.2%). Conclusion: The components of essential oil from S. fruticulosa were identified for the first time, and these substances may be responsible for the biological effects of these essential oils.

New Rare Ent-Clerodane Diterpene Peroxides from Egyptian Mountain Tea (Qourtom) and Its Chemosystem as Herbal Remedies and Phytonutrients Agents.

Genus Stachys, the largest genera of the family Lamiaceae, and its species are frequently used as herbal teas due to their essential oils. Tubers of some Stachys species are also consumed as important nutrients for humans and animals due to their carbohydrate contents. Three new neo-clerodane diterpene peroxides, named stachaegyptin F-H (1, 2, and 4), together with two known compounds, stachysperoxide (3) and stachaegyptin A (5), were isolated from Stachys aegyptiaca aerial parts. Their structures were determined using a combination of spectroscopic techniques, including HR-FAB-MS and extensive 1D and 2D NMR (1H, 13C NMR, DEPT, 1H-1H COSY, HMQC, HMBC and NOESY) analyses. Additionally, a biosynthetic pathway for the isolated compounds (1–5) was discussed. The chemotaxonomic significance of the isolated diterpenoids of S. aegyptiaca in comparison to the previous reported ones from other Stachys species was also studied.

Antioxidant and protective effect of Stachys pilifera Benth against nephrotoxicity induced by cisplatin in rats.

The aim of current study was to assess the antioxidant and renoprotective effects of Stachys pilifera Benth (S.P.B.) hydroalcoholic extract on nephrotoxicity induced with cisplatin (CP). Adult rats with bodyweight of 180-220g were divided into five groups (n=7) treated as follows: group 1, control; group 2, CP; group 3, pretreatment with S.P.B. before CP; group 4, posttreatment with S.P.B. after CP; and, group 5, S.P.B. extract. A single dose of CP (7mg/kg) was intraperitoneally injected on the fifth day and hydroalcoholic extract of S.P.B. (500mgkg-1 day-1 ) was orally administered. The levels of oxidative stress markers, biochemical tests, and histopathological staining were assayed in serum and renal tissue. Also, the chemical composition of S.P.B. extract was determined by GC-MS analysis. The main compositions of S.P.B. extract identified by GC-MS analysis, were hexadeca-2,6,10,14-tetraen-1-ol, 3,7,11,16-tetramethyl (24.77%), thymol (14.1%), and linolenic acid (13.4%). In groups treated and pretreated with S.P.B., blood urea nitrogen, creatinine, malondialdehyde, and nitric oxide metabolite in serum as well as malondialdehyde and protein carbonyl content of kidney tissues were significantly decreased in comparison to CP group; in contrast, total thiol group showed a significant increase in treated group as compared to CP group. Furthermore, histological investigation indicated that treatment with S.P.B. improved renal damages induced by CP. The current study showed that S.P.B. hydroalcoholic extract improved the biochemical parameters and kidney function as well as restored antioxidant activity in CP-induced nephrotoxicity. However, it needs more investigations to define the mechanism of S.P.B. action. PRACTICAL APPLICATIONS: In different regions of Iran, Stachys is demonstrated by 34 species, out of which 13 are endemic, one of these endemic species is Stachys pilifera Benth (S.P.B.). The oil of S.P.B. is mainly consisted of cis-chrysanthenyl acetate, cis-chrysanthenol, spathulenol, β-caryophyllene, linalool, and terpinen-4-ol. Moreover, phytochemical studies have shown the presence of compounds such as diterpenes, phenylethanoid glycosides, saponins, terpenoides, and flavonoids in Stachys species. The aerial parts of S.P.B. are consumed as herbal tea to treat several disorders, for example, infections, asthma, and rheumatoid arthritis in Iranian folk medicine. The aim of current study was to evaluate the antioxidant and protective effects of S.P.B. hydroalcoholic extract on nephrotoxicity induced with cisplatin (CP). The current study showed that S.P.B. hydroalcoholic extract improved the biochemical parameters and kidney function as well as restored antioxidant activity in CP-induced nephrotoxicity. However, it needs more researches to define the mechanism of S.P.B. action.

Phenolic Content, Antioxidant Effects and Tyrosinase Inhibitory Activity of Extract of Some Stachys Species from Iran

Phenolic Content, Antioxidant Effects and Tyrosinase Inhibitory Activity of Extract of Some Stachys Species from Iran

Chemical composition of the essential oil and n-hexane extract of Stachys tmolea subsp. Tmolea Boiss., an endemic species of Turkey, and their mosquitocidal activity against dengue vector Aesdes aegypti

Stachys tmolea subsp. tmolea Boiss. is endemic to Turkey and is a species of the genus Stachys L. which is one of the largest genera of the family Lamiaceae with about 300 species. The aims of this study were to examine the chemical composition of the essential oil and n-hexane extract of S. tmolea subsp. tmolea as natural sources of insecticidal activity against the dengue vector, Aedes aegypti. Analysis of the essential oil by GC-FID and gas chromatography-mass spectrometry (GC-MS) systems identified hexahydrofarnesyl acetone (15%), viridiflorol (10%), hexadecanoic acid (7%) and 9-geranyl-p-cymene (6%) as major components. The volatile components of the n-hexane extract were extracted using headspace solid-phase microextraction (HS-SPME) and were analyzed using GC-MS. The principal constituents were 3,4-dimethyl decane (16%), 3-methyl-3-pentanol (15%), 2-methyl-2-pentanol (12%), 1,4-bis (1,1-dimethylethyl) benzene (12%), heptanal (10%), acetic acid (6%) and decane (4%). Bioassay of the n-hexane extract, at 5 µg/mosquito, produced 90% mortality against adult Ae. aegypti while the S. tmolea essential oil demonstrated 13% mortality. No larvicidal activity was observed both in essential oil and n-hexane extract. Further studies are needed to assess the adulticidal activity of the responsible compounds in the crude extract.

Determination of nutritive value and analysis of mineral elements for wild edible Stachys lavandulifolia vahl. var. lavandulifolia growing in eastern Anatolia

The genus Stachys (Lamiaceae) is represented by about 300 species found in the world. Turkey is one of the richest countries in Stachys diversity being represented by 83 species with a level of 48% endemism. In this study, Stachys lavandulifolia Vahl. was collected from Alacabük mountain of Tatvan in Eastern Anatolia. S. lavandulifolia Vahl. subsp. lavandulifolia, known as “dağ çayı, tüylü çay”, is widely consumed in Anatolia as herbal tea. Stachys species belong to one of the oldest medicinal plants that are used both for pharmaceutical purposes and in folk medicine; It is used for the treatment of gastrointestinal and respiratory disorders, and is known as an appetizer, carminative, stimulant, digestive, diuretic, and throat pain reliever. Wild plants gathered from nature are cheaper food and important for human health. Thus, in the present study, nutritional value and mineral composition of used parts of selected S. lavandulifolia Vahl. var. lavandulifolia was investigated.In laboratory analysis, total ash, % N, crude protein, crude fiber and pH were examined as nutritional value. Useful minerals (Ca, Cu, Fe, K, Mg, Mn, Na, P, S and Zn) and some heavy metals (Cd, Co, Cr and Pb) that are hazardous elements for livings were also determined. The total ash, total nitrogen and crude protein, pH and crude fiber content of the plants were identified as 7.00%, 1.31%, 8.03%, 6.53% and 38.40%, and respectively. Mineral analysis showed that the wild plants samples contained considerably high amounts of potassium (17.46g/kg), phosphorus (4.70g/kg), calcium (17.25g/kg), magnesium (2.47g/kg), iron (241.37mg/kg), manganese (22.95mg/kg) and zinc (18.56mg/kg). This work contributed to the nutritional properties of some wild plants, and the results may be useful for the evaluation of dietary information.

Phenolic ingredients and therapeutic potential of Stachys cretica subsp. smyrnaea for the management of oxidative stress, Alzheimer’s disease, hyperglycemia, and melasma

Woundwort plants (Stachys species) are used traditionally as food additives, herbal tea, and medicine. At this work, antioxidant activity of S. cretica subsp. smyrnaea was comprehensively investigated using 6 different methods together with phenolic profile and total phenolics/flavonoids content of its several extracts. Moreover, key enzymes inhibitory potential of the herb associated with Alzheimer’s disease, hyperglycemia, and hyperpigmentation was evaluated. Promising antiradical activity (1.9–17.3 mg Trolox equivalents/g dry plant) and metal ion reducing power (4.6–14.6 mg Trolox equivalents/g dry plant) was recorded. Also, S. cretica manifested strong antityrosinase (2.45 mg kojic acid equivalent/g dry plant) and extraordinary anti-amylase and anti-glucosidase (6–61 and 14–47 mg acarbose equivalent/g dry plant, respectively) effects. Reverse phase high performance liquid chromatography analysis identified 12 phenolic compounds of them, chlorogenic acid, hesperidin, kaempferol, apigenin, and benzoic acid were the most abundant (426, 407, 390, 216, and 188 μg/g dry plant, respectively). Findings indicated that, S. cretica has great potential for possible applications in cosmeceutical, pharmaceutical, and food industries.

Gas chromatography combined with mass spectrometry and flame ionization detection for identifying the organic volatiles from Stachys arvensis, S. marrubiifolia and S. ocymastrum

Comparative analysis on the chemical composition and biological properties of the volatile fractions obtained from three Stachys species (S. arvensis, S. marrubiifolia and S. ocymastrum) growing in Tunisia is reported, for the first time, in the present work.The volatile organic compounds were extracted by hydrodistillation in a Clevenger-type apparatus, then analyzed using CG-FID and CG-MS. 37, 34 And 39 components were identified and quantified in previously mentioned species, respectively. It’s noticeable that more than 30% of the volatiles constituting each species are hydrocarbons. Taking into account only the identified compounds, the terpenes content in three Stachys species is lower than 20%. In addition, the comparative study of the chemical composition of these taxa showed that S. ocymastrum volatile fraction was characterized by its richness in phenlypropanoide (E)-anethol (18.1%) and hexahydrofarnesyl acetone (8.9%) in comparison with the two-other species. Moreover, the volatile fractions of S. arvensis and S. marrubiifolia showed a similarity in their chemical compositions.The antimicrobial activities were investigated using the micro-well plate dilution assay for five bacterial strains (Escherichia coli, Salmonella enterica, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis), for Candida albicans (ATCC 30031) yeast and the agar incorporation method for Aspergillus niger and Penicillium Spp. filamentous fungi.The results have shown significant antimicrobial activities against all tested bacteria and Candida albicans human pathogenic yeast. The minimal inhibitory concentrations, MIC values ranged from 156 to 625 μg mL−1. However, the volatile fractions presented a very low antifungal effects against Aspergillus niger and Penicillium spp.

Volatile Constituents of Three Stachys L. Species from Turkey

Volatile compounds of three Stachys species; Stachys macrantha (C. Koch) Stearn , Stachys sylvatica L. and Stachys annua ssp. annua var. annua L. were analyzed by a solid phase micro extraction (SPME) method with gas chromatography-mass spectrometry (GC-MS). A total of 38 (99.2%), 38 (99.0%) and 32 (98.8%) volatile compounds were identified with different constituents from S. annua ssp. annua var. annua, S. sylvatica and S. macrantha respectively. The major volatile constituents of the investigated three Stachys species were; α -pinene (11.2%), p -cymene (18.2%), carvacrol (28.8%) in S. macrantha , γ -muurolene (10.2%), α -cedrene (11.2%), limonene (37.0%) in S. sylvatica and α -pinene (11,4%), β -pinene (23.1%), (Z)-β -ocimene (24.8%) in S. annua ssp. annua var. annua. Comparison studies of all three Stachys species showed the high percentage of monoterpene hydrocarbons within 65.8%, 49.8% area in S. annua ssp . annua var. annua and S. sylvatica respectively. And oxygenated monoterpenes were the main group of constituents (42.1% area) of S. macrantha.

Composition of the essential oil of Stachys sericantha, S. gaziantepensis, and S. mardinensis (Lamiaceae) from Turkey

ABSTRACTThe genus Stachys L. one of the largest genera of Lamiaceae and Stachys species are used by many people for the therapeutic value of their essential oils. In this study, water-distilled essential oil of three species of Stachys was analysed. The analysis was performed by using gas chromatography-flame ionisation detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS) systems simultaneously. The major components found were hexadecanoic acid (23.7%), dodecanoic acid (11.3%), and caryophyllene oxide (10.7%) for S. sericantha P.H. Davis (endemic); α-pinene (53%), β-pinene (8.2%) for S. gaziantepensis M. Dinc and S. Doğu (endemic); Menthyl acetate (15.3%), isomenthone (15.0%), pulegone (10%), spathulenol (7.0%), and caryophyllene oxide (6.7%) for S. mardinensis (Post) R. Mill.

Polar Constituents, Essential Oil and Antioxidant Activity of Marsh Woundwort (Stachys palustris L.).

Stachys palustris, well-known as marsh woundwort, is a perennial herb growing in wet environments of Europe. Its tubers, leaves and seeds are eaten raw or cooked. Alike other Stachys species, the plant is also used as a traditional remedy. Despite S. palustris has been consumed for centuries, little is known about its chemical constituents. In this work the main secondary metabolites of S. palustris from Hungary and France have been analysed. From the plant ethanolic extracts, ethanoid glucosides, isoscutellarein derivatives, caffeoyl-quinic acids and iridoids have been isolated and structurally characterized by NMR. The essential oils were analysed by GC/MS and showed (E)-phytol, fatty acids and carbonylic compounds as the most abundant compounds. The radical scavenging capacity of plant ethanolic extracts, as evaluated by the DPPH assay, was noteworthy, with IC50 values of 92.08 - 105.42 μg/ml.

391 - Evaluation of Biological Activities of Three Stachys Species from Turkey

Stachys L. genus belongs to Lamiaceae which is represented by approximately 300 species and has a wide distribution in the world, except Australia and New Zealand. Stachys species are traditionally used for the treatment of infection, asthma, wounds, tumors, rheumatic and inflammatory diseases, cough, and ulcers. In this study it is aimed to evaluate acetylcholinesterase (ACE), butyrylcholinesterase (BCE), cyclooxygenase COX-1 and COX-2, and also monoamine oxidase MAO-A and MAO-B enzymes inhibitory effects of the aqueous methanolic extracts of S. annua L. subsp. annua var. annua, S. iberica Bieb. subsp. iberica var. densipilosa and S. byzantina C. Koch. at 250 µg/mL. Total phenolic contents of the extracts were determined by Folin-Ciocalteu assay. The enzymatic activity results showed that S. iberica extract exhibited moderate COX-1 inhibition (45.25%) whereas S. annua and S. byzantina extracts showed weak effects when compared to Celecoxib (94.05%). With regard to the MAO-A inhibitory activity, Stachys extracts were significantly effective in the order: S. annua (72.65%) > S. iberica (50.85%) > S. byzantina (49.23%) when compared to Clorgiline (84.75%). Also S. annua extract had the most and strong effect on MAO-B inhibition (65.98%). The extracts showed slight inhibitory activity on ACE and BCE enzymes. Between the three Stachys extracts, S. byzantina had the highest total phenolic content as 151.64 mg gallic acid equivalent/100 g plant.

Evaluation of antioxidant activity, total phenolics, total flavonoids and LC–MS/MS characterisation of phenolic constituents in Stachys lavandulifolia

The aim of this study was to evaluate phenolics profile and antioxidant activity of Stachys lavandulifolia. Total phenolics (TP), total flavonoids (TF), DPPH• assay (IC50), ferric ion reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) of the methanolic extract were measured. The content of TP, TF, IC50, FRAP and TAC, were obtained as 16.59 gallic acid equiv./g dry matter (DM), 4.48 mg quercetin equiv./g DM, 2.07 (μg/mL), 0.014 (absorbance/mg phenolic) and 14.61 (mg BHT equiv./g DM), respectively. The results showed that S. lavandulifolia, compared to other species of Stachys, had moderate TP content with desirable antioxidant activity. Subsequently, 59 various phenolic compounds were identified and confirmed in the methanolic extract of S. lavandulifolia using high mass accuracy by MS2 experiments. The compounds consisted of 6 hydroxybenzoic acids and hydroxybenzoic aldehydes, 9 hydroxycinnamic acids, 1 coumarin, 32 flavonoids, 3 lignans, 2 stilbenes, 3 tannins and 3 other phenolics.

Potential sources for the management global health problems and oxidative stress: Stachys byzantina and S. iberica subsp. iberica var. densipilosa

IntroductionStachys species have been applied externally for the treatment of wounds and used internally for abdominal pains, cramps, dizziness, fever, gout and menstrual disorders. This study aimed to evaluate the in vitro antioxidant and enzyme inhibitory activities of solvent extracts of Stachys byzantina and S. iberica subsp. iberica var. densipilosa. MethodsTwo Stachys species were analyzed for their antioxidant activities using different test systems and, inhibitory properties against acetylcholinesterase, butrylcholinesterase, tyrosinase, α-amylase and α-glucosidase investigated. Phenolic contents in the extracts were determined by HPLC. ResultsIn S. iberica subsp. iberica var. densipilosa, the amount of phenolics, flavonols and saponins were found to be slightly higher than that of S. byzantina. However, the ethyl acetate, methanol, and water extracts of S. byzantina contained considerable amount of flavonoid compounds (31.99, 47.70, and 34.66mg REs/g extract, respectively). The ethyl acetate extracts were found to be rich in phenolics, chlorogenic acid was the most abundant compound. Apart from the phosphomolybdenum assay, S. byzantina exhibited higher antioxidant activity than that of S. iberica subsp. iberica var. densipilosa. The methanol extract of this species showed remarkable activity in phosphomolybdenum, radical scavenging (on ABTS and nitric oxide radicals), and CUPRAC assays (1.49mmol TEs/g extract, 143.85mg TEs/g extract, and 1.84mmol TEs/g extract, respectively), whereas the water extract exhibited a promising activity in radical scavenging (on DPPH and superoxide radicals), reducing power (FRAP), and chelating effect assays. In general, ethyl acetate extracts of both species showed remarkable inhibitory activities on the enzymes. Ethyl acetate extract of S. iberica subsp. iberica var. densipilosa showed higher inhibitory activity on acetylcholineserase, butrylcholinesterase, α-amylase than the same extracts of S. byzantine (2.16, 4.20mg GALAEs/g extract, and 0.34mmol ACEs/g extract, respectively). ConclusionsResults suggest that the natural compounds found in S. byzantine and S. iberica subsp. iberica var. densipilosa extracts may provide some evidence to confirm their traditional use for the management of certain health problems such as Alzheimers disease, skin disorders, diabetes mellitus and when oxidative damage occurs.

Tipificaciones en especies sudamericanas de Hedeoma, Lepechinia y Stachys (Lamiaceae) y nuevo sinónimo en Lepechinia

Se designan ocho lectotipos para los siguientes nombres pertenecientes a especies de Lamiaceae: Astemon graveolens Regel, Hedeoma adscendens Rusby, Hedeoma mandonianum Wedd., Lepechinia floribunda (Benth.) Epling, Lepechinia sagittata Epling, Lepechinia vesiculosa (Benth.) Epling, Sphacele hastata Griseb. y Stachys albicaulis Lindl. A su vez se propone a Lepechinia sagittata Epling como nuevo sinónimo de Lepechinia floribunda (Benth.) Epling.

Genetic diversity and morphological variability of fifteen Stachys (Lamiaceae) species from Iran using morphological and ISSR molecular markers

AbstractStachys L. (Lamiaceae) exhibits a great range of variation with cosmopolitan distribution. An area extremely rich in taxa including 34 species has been introduced in Iran. This study documents the genetic diversity and morphological variations of 48 accessions from fifteen Stachys species and two subspecies belonging to six sections. All of the specimens were collected from natural habitats in Zagros region of Iran. In order to evaluate morphological variations, 21 characters were used. Eleven ISSR primers with DNA samples of leaves were applied to assess the genetic variations among species and populations of the Stachys genus. Based on the results, from the eleven primers screened, nine primers produced reproducible bands. The highest percentage of polymorphism included 100% in section Aucheriana. Analysis of molecular variance revealed a high proportion of gene variation (42%) among populations within species. In total sections, the UPGMA clustering dendrogram genetically detected different groups. Based on the morphological analysis, each section was definitely positioned. A high genetic diversity and polymorphism was found among Stachys accessions belonging to each section. Based on the findings, the germplasm diversity was due to the wealth genetic resources in the central Zagros region which are important in plant breeding and gene conservation purposes. In addition, morphological and ISSR molecular markers are useful to display the diversity in Stachys genus.

Composition and Antibacterial Activity of the Essential Oil of Phlomidoschema parviflorum from Iran

The monotypic genus Phlomidoschema Vved. belongs to the family Lamiaceae, subfamily Lamioideae and tribe Stachydeae [1]. Phlomidoschema parviflorum (Benth.) Vved. grows in restricted parts of Iran (Khorasan Province) as well as Afghanistan, west of Pakistan, Pamir Mountains, and Punjab of India. Morphologically, the species is characterized by perennial, multi-stemmed and subshrub habit and white subfloccose and canescent hairs on the leaves and stem [2]. The species is considered vulnerable and rare according to IUCN categories [3]. The infusion of the aerial flowering part of the plant is used locally for the treatment of gastrointestinal disorders, colic and stomachache. From the phylogenetic point of view, the genus is closely allied with the genus Stachys. The most relative and closest species of the genus Stachys to the genus Phlomidoschema was found to be S. lavandulifolia Vahl. [1]. As far as our literature survey could ascertain, there is no report on the oil composition and biological activity of P. parviflorum. So, in continuation of our research on the composition and biological activity of Iranian aromatic and essential oil-bearing plants of the mint family [4–10], we have investigated the composition and antibacterial activity of the essential oil of P. parviflorum. The aerial full flowering parts of Phlomidoschema parviflorum were collected from Khorasan-e Razavi Province, Ghayen towards Gonabad, Zirkuh region with an altitude of 1300–1350 m. A voucher specimen was identified and deposited (MPH-1220) in the Medicinal Plants and Drugs Research Institute Herbarium (MPH), Shahid Beheshti University of Tehran, Iran. Air-dried aerial parts (200 g) were subjected to hydrodistillation for 3 h using a Clevenger-type apparatus. The oil was dried over anhydrous sodium sulfate and stored in sealed vials at 4C until analysis and tested. GC analysis was carried out on a Thermoquest-Finnigan Trace GC instrument equipped with a capillary DB-5 fused silica column (60 m 0.25 mm i.d., film thickness 0.25 m). The oven temperature was raised from 60C to 250C at a rate of 5C/min, then held at 250C for 10 min. Nitrogen was used as the carrier gas at a flow rate of 1.1 mL/min; the split ratio was adjusted to 1:50. The injector and detector (FID) temperatures were kept at 250C and 280C, respectively. GC-MS analysis was carried out on a Thermoquest-Finnigan Trace GC-MS instrument equipped with a DB-5 fused silica capillary column. Helium was used as the carrier gas at a flow rate of 1.1 mL/min with a split ratio of 1:50. The quadrupole mass spectrometer was scanned over 45–465 amu with an ionizing voltage of 70 eV. Gas chromatographic conditions were the same as given above for GC. The constituents of the oil were identified by calculation of their retention indices under temperature-programmed conditions for n-alkanes (C 6 –C 24 ), and the essential oil on a DB-5 column. Identification of individual compounds was made by comparison of their mass spectra with those of the internal reference mass spectra library or with authentic compounds and confirmed by comparison of their retention indices with those reported in the literature [11]. In vitro antibacterial activity of the essential oil was assessed against Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 as models of Gram positive and Gram negative bacteria, respectively. The broth microdilution susceptibility method using 96 well trays was used to determine the minimum concentration of essential oil required for inhibition of visible growth (MIC) or killing (MBC) of the test strains. For this purpose, the standard protocol of CLSI (Clinical Laboratory and Standards Institute) was used with some modifications [12]. The inoculants of the bacterial strains were prepared from freshly cultured strains using sterile normal saline, which were adjusted to 0.5 McFarland standard turbidity and then further diluted (1:1000 for bacteria) by sterile Mueller-Hinton broth (MHB) just before adding to the wells containing a desired range of diluted essential oil.

Flavonoid Patterns and their Diversity in ten Stachys L. (Lamiaceae) Species from Iran

Stachys genus with medicinal properties and high polymorphic features has been considered oneof the largest genera of Lamiaceae. The aim of this study was to determine the flavonoid patternvariations and flavonoid groups in ten Stachys species belonging to two sections; Fragilicaulis,and Aucheriana. The studied species were collected from natural habitats in Iran and analysedfor their flavonoid constituents using thin layer chromatography with silica gel. The purificationof the flavonoid compounds of each species was carried out using column chromatography withsephadex LH20. The identification of flavonoid class was confirmed by spectral data. In orderto study the flavonoid variations, cluster analysis was used with SPSS ver.20 software. Theresults of this study showed that the highest variations were found in Stachys pilifera Benth.,Stachys aucheri Benth., Stachys ballotiformis Vatke and Stachys benthamiana Boiss. Based onthe results, six flavonoid classes were identified. Most of the flavonoid classes were found to beflavones. The flavones and isoflavones were observed in section Fragilicaulis and flavanones,flavonols, isoflavones, dihydroflavonol, chalcones and flavones were in section Aucheriana. Itcan be concluded that the flavonoid compounds are appropriate markers in chemotaxonomicstudies of the Stachys genus.

Carthamus, Salvia and Stachys species protect neuronal cells against oxidative stress-induced apoptosis

Context: Finding effective therapies for neurodegenerative diseases is of utmost importance for the aging population. Plants growing in Iran are rich sources of antioxidants and active phytochemicals.Objective: The protective capacity of plants, with a special focus on those with reported antioxidant or neuroprotective potential or nervous system-related applications in folk medicine, was tested against oxidative stress-induced apoptosis.Materials and methods: Aerial parts of 20 plants including Carthamus, Salvia, and Stachys species were extracted with 80% methanol and dichloromethane and preincubated with neuronal PC12 cells for 3 h. Oxidative stress and apoptosis were induced by hydrogen peroxide (75 µM, 1 h exposure). Cell viability and intracellular reactive oxygen species (ROS) were measured by MTT and 2′,7′-dichlorofluorescein-diacetate (DCFH-DA) assays, respectively, while apoptosis was determined by annexin V-FITC/propidium iodide staining by a flow cytometer.Results: Eighty percent methanol extracts of Carthamus oxyacantha Bieb. (Asteraceae), Salvia santolinifolia Boiss. (Lamiaceae), and Salvia sclarea L. (Lamiaceae) at the concentration of 100 μg/ml showed significant neuroprotection in the MTT assay by 38.7, 34.7, and 39.5%, respectively, and inhibited intracellular ROS by 48.6, 61.9, and 61.4%, respectively. The first two extracts also significantly inhibited apoptosis. Dichloromethane extracts of C. oxyacantha and Stachys pilifera Benth. (Lamiaceae) at the concentration of 25 μg/ml showed neuroprotection by 27.5 and 26.5%, respectively, and inhibited ROS by 44.5 and 39.4%, respectively.Conclusion: The above-mentioned plants seem to have important biological activities and their further study may lead to the discovery of new natural therapeutics useful against disorders such as Alzheimer and Parkinson diseases.