[Quality and quantity determination of brassino-steroid hormones by HPLC and related assay in some supplement capsules]

Background: Brassino-steroids as hormonal plants are of the most important components that have shown the biological effects. These compounds may be act as inducer of growth in the plant and their concentrations is increased under some stress conditions. Such clinical studies show that these hormones have also anabolic activities similar to the human anabolic hormones hence, may be administrated as supplements in athletes. Their increasing consumption as supplement suggests introducing a precise and accurate analytical method for determination of brassinosteroids in pharmaceutical dosage forms and also in some biological fluids

Objective: In the current study an HPLC method for detect and determination of brassino-steroids in capsules was developed and optimized in aspects of analytical conditions

Methods: In order to analysis of the sample a reversed phase HPLC system including a C18 column, and a mobile phase including water: Acetonitrile in an acidic media were used. Detection was carried out at 210 nm by an UV detector

Results: In this study, we attempted to optimize some analytical aspects e.g: solvent, stationary phase and other assay parameters to obtain best condition to assay. Also the validation parameters such as LOD, LOQ, accuracy, precision and linearity of the method were also studied and were satisfactory

Conclusion: Obtained data indicated that HPLC is a suitable analytical method for assay of brassino-steroids in pharmaceutical dosage forms

Effect of ethanolic extract of mespilus germanica on cutaneous leishmaniasis in BALB/c Mice

Leishmaniasis is a vector-borne disease caused by flagellated protozoan parasites of the genus Leishmania, which affects both humans and other mammals. Most of the available drugs against the disease are toxic and some are parasite resistance to them. The purpose of this study was to examine the effect of Mespilus germanica extract on cutaneous leishmaniasis [CL] in BALB/c Mice. Ethanolic extract of Mespilus germanica with 40, 60 and 80% concentrations were prepared. Then, the BALB/c mice were inoculated subcutaneously by 0.1 ml liquid phase culture containing promastigotes of Leishmania major. Ethanolic extract of the leaves of Mespilus germanica in different concentrations, were used topically on CL lesions. The mean diameter of the lesions were decreased, and also the number of parasites in the lesions had declined with complete healing by ending the period time of treatment in 4 mice [26.7%], [p<0.05] and in 9 animals [82%], [p<0.05] respectively, by using the 40% concentration of the extract. Also in a concentration of 60%, mean ulcer diameter decreased, with complete healing in 3 mice [20%], [p<0.001]. In this concentration, the mean number of parasites in lesions had declined [66.4%], with total elimination in 8 animals [p<0.001]. We showed that the extract of Mespilus germanica has the highest effectiveness in concentration of 40%, causing greater reductions in both ulcer diameter and the number of parasites in the lesions compared with other prepared concentrations. Therefore, we suggest the use of 40% extract for the treatment of human cases

[Optimization of extraction method of hyoscine from Hyoscyamus niger L.]

Hyoscyamus niger L. from Solanaceae is an important pharmacological plant which contains tropane alkaloids, tropane alkaloids constitute one distinct group of secondary metabolites of the Solanaceae. Hyoscyamus species are rich source of tropane alkaloids, mainly hyoscyamine and hyoscine [scopolamine], which are widely used for their mydriatic, antispasmodic, anticholinergic, analgesic and sedative properties. Therefore determine of optimized method is necessary for extraction of hyoscine from Hyoscyamus niger L. The purpose of this research is determine of optimized conditions for extraction of hyoscine from Hyoscyamus niger L. Three factors, temperature, extraction time and mesh size of plant were studied using three level box-behnken design, the hyoscine content were measured by Gas chromatograph, Younglin Acm 6000 model. The results showed that the best conditions for extraction of hyoscine from Hyoscyamus niger L. is Ultrasonic extraction in 43 C, 130 min and 45 mesh size. It can be concluded that in order to efficient extraction of hyoscine from Hyoscyamus niger L., special conditions should be considered

Chemical composition and antimicrobial activity of Oliveria decumbens volatile oil from west of Iran

Oliveria decumbens Vent. is an endemic plant of Flora Iranica that grows in high temperature areas of south and west of Iran. It is used for indigestion, diarrhea, abdominal pain and fever in traditional medicine. In this investigation, chemical composition and antimicrobial effects of Oliveria decumbens has been studied. The essential oil of aerial parts of Oliveria decumbens was obtained by hydrodistillation and analyzed by GC-MS. The antimicrobial activity of the essential oil was also investigated against three Gram positive bacteria, two Gram negative bacteria and two fungi using disc diffusion method. 10 components were identified in the essential oil of Oliveria decumbens. The main components were gamma-terpinene, myristicin, thymol, p-cymene and carvacrol. The essential oil showed high antimicrobial activity against Gram-positive bacteria, Escherichia coli, Aspergilus niger and Candida albicans but low antibacterial effect against Pseudomonas aeroginosa. High amount of thymol, carvacrol and p-cymene in Oliveria decumbens essential oil may be a reason of antimicrobial effects of the plant and the plant essential oil can be considered as a natural source of preservatives in food and cosmetic industries

[Effects of essential oil and extract of thymus vulgaris, zataria multiflora and eugenia caryophilata on Vero, Hela, HepII cell lines by MTT assay]

Natural flavors essential oils are almost present in various food stuffs, cosmetic and pharmaceutical products. Safety assessment of these food additives should be seriously considered as long as their due wide applications are favorable. Natural flavors essential oils are widely used in various foods, cosmetic and pharmaceutical products. These kinds of additives are applied as colors, preservatives, aroma, antioxidant agents and tasting agents. In some cases, it may happen that the large use of some of these additives may accompany with some concerns and unwanted toxic disturbs. In this study, Concentrations of 24, 12, 6, 3, 1.5, 0.75, 0.375, 0.1875ppm [micro g/ml] of three essential oils, Eugenia caryophyllata, Thymus vulgaris and Zataria multiflora, were tested in MTT assay with three cell line Vero, Hep2, Hela. All three Essential oils and extracts were shown cytopathological effects on three cell lines in MTT assay in at least 0.04 micro g/ml to maximum 3.0 micro g/ml concentrations. Based on obtained results, all studied essential oils and extracts may have cytopathologic effect in specific concentrations so suggested using of this compound should be performed with considering their proper concentration and more safety studies on them

Synthesis and radioprotective study of novel amino-alkyldithiocarbamic acid derivatives against gamma-irradiation in mice

The aim of this study was to evaluate the radioprotective capacity of some novel aminoalkylated dithiocarbamic acid potassium salts against gamma-irradiation in mice. Eight compounds containing 2-aminoethyl-, 3-aminopropyl-, 4-aminobutyl-, 5-aminopentyl-, 6-aminohexyl-, 7-aminoheptyl-, 8-aminooctyl and 9-aminononyl of dithiocarbamate derivatives were prepared. Male NMRI mice were injected intraperitoneally [IP] with a geometric progression of doses [300-1000 mg/kg], through the dose response range for lethal toxicity. To evaluate the radioprotecive activity, one-half of the toxic LD[50] of each compound were injected IP to groups of twenty mice, 30 minutes prior to gamma-irradiation. The treated animals were kept for 30 days, and the lethality was recorded each day. Among eight compounds of alkyl dithiocarbamic acid derivatives, 5-aminopentyl, 7-aminoheptyl, 8-aminooctyl and 9-aminononyl dithiocarbamic acid mono potassium salts are new compounds. All evaluated compounds showed a concentration dependent effect on the survival in mice. The LD[50] values were found to be more than 599 mg/kg. The percentages of 30-day survival of mice for 2-aminoethyl, 7-aminoheptyl and 8-aminooctyl dithiocarbamic acid derivatives were 7%, 40% and 13.5%, respectively, when injected 30 minutes before gamma-irradiation. Other compounds had no radioprotective effects. Statistical analysis showed a significant difference between the treated and control groups for the 7-aminoheptyl derivative [p<0.05]. Among the compounds investigated in this study, 7-aminoheptyl dithiocarbamate derivative showed more radioprotective effects in comparison with the others. Although it seems that the radioprotective effects in these derivatives correlate with the size of the alkyl chain, more experiments are required to support this hypothesis

[Investigation of the best method for extraction of phenolic compounds from echinaceae purpurea L. [moench]]

Echinacea spp. Have been used in traditional medicine for many years. Recently, many investigations have been done on this plant and they showed that the plant is an immunostimulant agent. The plant contains different components such as flavonoids, phenolic compounds and isobutylamides. Phenolics are one of the most important agents in immunostimulant effects of the plant; therefore, determination of a method for the best extraction of these compounds from the plant is important. In this research, the effects of solvent, extraction procedure, plant particle size and plant: solvent ratio on extraction of phenolic compounds of Echinacea purpurea [L.] Moench have been investigated. Different polar and non-polar solvents from hexane to acidic water were used in order to determine the best solvent for phenolics extraction. Moreover, the methods of maceration, sonification, perculation, digestion and continuous extraction [by use of soxhelet apparatus], different particle sizes of the plant powder and various plant: solvent ratios were examined as well. The results showed that the best method for extraction of phenolics is methanol: water 80:20 as solvent, method of digestion [2h, 50[degree sign] C], particle size of 300 micro m for the plant powder and plant :solvent ratio of 1:200. It can be concluded that in order to efficient extraction of phenolic compounds from Echinacea purpurea aerial parts, special condition should be considered

Fingerprint study of Thyrnus spp.aby TLC

Thymus is a widely used medicinal plant in food and pharmaceutical industries. Among different species of Thymus, T. vulgaris is used more than other species in therapeutic dosage forms. This study was conducted to differentiate various Thymus species by TLC fingerprint. In order to identify and differentiate various species of Thymus, TLC fingerprints of essential oil, dichloromethane, ethylacetate and n-butanol extracts of four available species named T. vulgaris, T. pubescens, T. citriodorus and T. daenensis were obtained and compared with each other. The results showed that the TLC chromatograms of essential oil and n-butanol extract cannot be used as differential identification for the above-mentioned species; while the ethylacetate and dichloromethane extracts are more reliable to be used for TLC fingerprints. TLC-chromatogram of ethylacetate extract is specific for identification of T. pubescens and T. vulgaris while T. citriodorus and T. daenensis can be best identified by their TLC fingerprints obtained from dichloromethane extract. For best differentiation of various Thymus species, different extracts of the plants should be used

effects of fruit maturity and 1-methylcyclopropene [1-MCP] treatment on alpha-farnesene metabolism in scald resistant and susceptible cultivars of apple fruit

It was found that, climatic conditions and fruit ripeness are important factor for the occurrence and intensity of superficial scald. 1-MCP is an inhibitor of ethylene action in plants and inhibits many ripening related processes along with the production of ethylene and volatiles. Apples are often held for several months at low temperature either in air [MA] or in controlled atmosphere [CA] storage, therefore they are prone to the occurrence of a large number of physiological disorders. In this work an investigation was made in order to determine the effect of 1-methylcyclopropene [1.0 microl I [-1]] treatment on superficial scald development and alpha farnesene metabolism on scald-resistant 'Golden Delicious' and scald-susceptible 'Imperial Delicious' apples. Fruits harvested on two dates and stored 24 weeks in refrigerated air storage [RA] at 6.5-1 °C, Peel tissue samples were taken at harvest after 6, 12 and 24 weeks. A reversed-phase high-performance liquid chromatographic procedure with UV detection at 232 nm and 269 nm has been used for the determination of alpha-farnesene and Ctols contents. Scald symptoms were restricted to untreated control fruit of both harvests of Imperial as during storage period increased to 65 and 45%, respectively for the first and second harvest compared with no scald symptoms in those in Golden Delicious. At 1.0 microl I [-1], 1-MCP completely eliminated superficial scald during storage period in Imperial apples. Large increases in peel tissue content of alpha-farnesene observed in untreated fruit of both harvest dates in Golden Delicious and Imperial cultivars. 1-MCP strongly decreased alpha-farnesene and conjugated trienols accumulation in treated fruit peel tissues at both maturity stages of two cultivars during storage. The results confirm the proposed role of Ctols in scald induction and indicate that alpha farnesene production is strongly regulated by ethylene. In general, we conclude that only alpha-farnesene synthesis and its oxidation products [Ctols] levels in peel tissue could not be involved to the inherent nature of scald susceptibility or resistance in two cultivars tested

Effect of Allicin of garlic on production nitric oxide of macrophage to Candida albicans

Garlic is an herbal plant having various compounds. one of the most important of which is allicin with antibiotic property. Candida Albicans is opportunistic yeast which in case of immune system dysfunction is considered as a pathogenic agent. Immune mechanisms against this fungus are Macrophages which act with oxidative and non-oxidative fungicide mechanisms. Oxidative mechanism includes active oxygen and nitrogen mediators which are produced by active macrophages and thereby the microorganisms are eliminated. The aim of this study was to investigate Allicin effects on the increase activites macrophage to product of Nitric Oxide It is examined in this study the effect of garlic allicin on macrophages' activity in releasing Nitric oxide against Candida Albicans. Garlic allicin was prepared by the method of chloroformic extract, and then made to react with macrophages from male mouse Balb/c with 2-8 week of age in vitro. Candida Albicans was divided into two groups: with and without allicin. Both cases compared as positive and negative samples, the rate of macrophages' activity was determined through production of Nitric Oxide. After examining the rate of Nitric Oxide produced by macrophages, results showed that allicin as a natural material activates immune system against this fungus, so that macrophages with allicin can produce more Nitric Oxide than the group without allicin. This was obtained by comparing the results from these two groups and the control group. Regarding the important role of Candida Albicans in Candidiasis and applying the preventing agents by this fungus in suppressing the macrophages' activities in producing Nitric Oxide, and also by studying related literatures, we concluded that allicin extracted from garlic can affect greatly on the macrophages' activities in producing Nitric Oxide against agents of Candidiasis disease

Taxane diterpenoids from Taxus baccata L. growing in Iran

There are eight Taxus species and two hybrids in the world and Taxus baccata L. [European yew] is the single representative in Iran. Until now, a large number of taxoids possessing different skeleton systems, as well as lignans, flavonoids, steroids and sugar derivatives have been isolated from various Taxus species. Taxoids are highly oxygenated diterpenes isolated from different species of yew trees [family Taxaceae]. They have received considerable attention after the discovery of the clinical efficacy of paclitaxel against different cancers due to its remarkable inhibition of microtubules polymerization leading to apoptosis of cancer cells. This study was conducted for extraction, purification and identification of taxoids occurring in the aerial parts of Taxus baccata L. growing in Iran. The plant materials were extracted with organic solvent and after defatting, partitioning, column chromatography and thin layer chromatography steps, the obtained compounds were characterized on the basis of spectral data. Two taxane diterpenoids were isolated. The structures of these taxoids were established as 5-Cinnamoyl-10-acetyltaxicin-I and 2-Deacetyltaxinine E on the basis of spectral analysis. These two taxoids were not previously encountered in Taxus baccata L. aspecies

Mutagenicity of four natural flavors: Clove, cinnamon, thyme and Zataria multiflora Boiss

Natural flavors are widely used in various foods, cosmetic and pharmaceutical products. These kinds of additives are applied as colors, preservatives, aroma and tasting agents. The large-scale use of certain food flavors requires accumulation of toxicological data on these substances, particularly in cases where structural similarities with other known substances showing genotoxic or carcinogenic properties indicate that some restrictions on human consumption or exposure should be implemented the case of the flavors. In this study, Concentrations of 50, 100, 200, 300, 500, 1000 and 2000 micro g/ml of four essential oils, Eugenia caryophyllata [Clove], Cinnamum zeylanicum [Cinnamon], Thymus vulgaris [Thyme] and Zataria multiflora, were tested in Salmonella typhymurium strains TA100 with and without rat liver S9 using Ames Salmonella reversion assay. Without S9 fraction, increase in mutant colonies per plate was not observed in all used concentrations. Also with S9 fraction all of samples had no significant increase in mutant colonies per plate except Clove in 500 micro g/ml, and higher concentrations. Based on obtained results, Clove oil may have mutagenic effect in 500 micro g/ml and higher concentration

Investigation of a and beta - thujone content in common wormwood from Iran

Artemisia absinthium L. [common wormwood] has been widely used in traditional medicine. The plant contains toxic components: a and beta-thujone, therefore, it should be used with caution. Since common wormwood is considered to be a toxic plant, we decided to evaluate toxic compounds quantities in those used in Iranian folk market. Five samples were purchased from Tehran markets. Essential oil of the flowers was obtained by water distillation. Essential oils were injected to gas chromatograph and their thujone content were determined according to the calibration curve. The results showed that alpha-thujone is absent in all samples and p-thujone content in the essential oils is much less than its amount reported from other countries. Since the quantity of toxic compounds are low in the plant growing in Iran, it is proposed that Artemisia absinthium L. can be studied for its some medical uses

Cytokinins affect fermentation product accumulation, vitamin c reservation and quality of stored broccoli in modified atmosphere packages

Broccoli inflorescences are rich in health promoting compounds such as vitamin C which may contribute to the high antioxidant capacity found in freshly- harvested broccoli. However, high looses of this essential compound has been reported after harvesting. Modified atmosphere packaging has been shown to be potentially useful in storability and quality retention of Broccoli florets; however, supplemental methods of extending shelf life are desirable because of the high fermentation product. In this way florets were treated post harvest with benzyl adenine at 50 ppm before packaging in polymeric bags [polyethylene and polypropylene] and storage at 1°C. Factors including ethylene production, chlorophyll, vitamin C, fermentation product, appearance, pH and titrable acidity were measured every three day intervals. During storage of cytokinin treated florets under MAP, all changes related with loss of quality were reduced and delayed with time. Additionally vitamin C remained almost unchanged. This improved retention was mainly due to decreased ethylene production. Cytokinin treatment significantly reduced fermentation product in packaged broccoli

[Taxuspinanane G from the aerial parts of Taxus baccata L. growing in Iran]

The discovery of paclitaxel [Taxol] as a potent anticancer drug from Taxus brevifolia has encouraged several groups all over the world to conduct research work on other Taxus species, in order to isolate potentially more effective paclitaxel derivatives for the treatment of various cancers or as starting materials for semi-synthesis As a consequence, more than 400 taxane-type diterpenes have been isolated from various Taxus plants, and some of them were found to possess interesting anticancer activity. This study was conducted for isolation, purification and identification of taxoids occurring in the aerial parts of Taxus baccata L. from Iran. The aerial parts of the plant were extracted with organic solvent and after defatting, partitioning, column chromatography and thin layer chromatography steps, the isolated compound was identified on the basis of NMR spectrum data. Taxuspinanane G was isolated and identified according to the NMR spectrum data. In this study, taxuspinanane G was isolated for the first time from the aerial parts of Taxus baccata L. This is the first report of the isolation of this compound from the given species

Chemical composition and antimicrobial activity of Artemisia annua L. essential oil from Iran

The composition of the essential oil obtained from the dried flowering aerial parts of Artemisia annua L. [Compositae] was analysed by GC and GC/MS. Thirty-two components were identified in the essential oil of A. annua L. with campher [48.00%], 1,8-cineole [9.39%], camphene [6.98%] and spathulenol [4.89%] as major components. The essential oil was evaluated for antibacterial and antifungal activities. The activity was more pronounced against fungal organisms than against Gram-positive and Gram-negative bacteria

Comparison of super critical fluid extraction and hydrodistillation methods on lavander's essential oil composition and yield

Lavender's essential oil is commonly used in aromatherapy and massage. Its major clinical benefits are on the central nervous system. Linalyl acetate and linalool are the most predominant chemical constituents in the essential oil of Lavandula angustifolia Comparison of super critical fluid extraction [SCFE] and hydrodistillation [HD] methods on Lavander?s essential oil composition and yield methods In this work we extracted essential oil of this plant with two different methods; SCFE and HD and further analyzed by GC and GC-MS method. seventeen compounds were identified in the oil which prepared by HD, the major components of them were j. pinene [35.9%] and lavandulyl acetate [14.1%]. In the SCFE extracted sample, the most frequent components were linalyl acetate [73.5%] and lavandulyl acetate [7.5%]. According to obvious difference in the composition of the essential oils prepared by two different methods [SCFE and HD], it seems that extraction method differ chemical composition of the oil and probably affects pharmacological properties

[Investigating on the quality of wild licorice roots collected from different regions of Iran]

Glycyrrhiza glabra L. has been used in respiratory diseases, gastric and duodenal ulcers and liver insufficiencies for many years. The plant grows in different regions of Iran. Since our country is considered one of the greatest exporting countries of the plant roots, investigation of licorice quality from different parts of Iran and determination of the best area for its growing seems to be important. According to validated references, glycyrrhizic acid percentage and water-soluble extractives are determinative of quality; therefore, in this investigation, these two factors have been measured in the plants collected from different regions of Iran. Roots of the plant were collected from Kerman, Kermanshah, Sirjan, Sarhadd [Fars province], Istahban [Fars province], Ghasredasht [Fars province], Najaf Abad [Isfahan province], Ganjnameh [Hamedan province], Ekbatan [Hamedan province], Mahabad, Khorram Abad and Ardabil and water-soluble extractives using maceration method and glycyrrhizic acid percentage with HPLC technique were determined in each sample. The results showed that the amount of water-soluble extractives and glycyrrhizic acid are high in samples from Kermanshah, Sarhadd and Kerman while these two factors are the lowest in the plant roots collected from Ekbatan and Ganjnameh. The plant roots collected from Kermanshah, Sarhadd and Kerman have the highest quality and are the best for medicinal purposes, sweetening and flavoring agent and for exporting as well

[Study of the process of essential oil extraction from rosemary plant by using steam distillation method]

Rosemary [Rosemarinus officinialis L.] is an aromatic, medicinal and condiment plant that belongs to the family labiatae, reaching a height of 1.5 meter. Essential oil of rosemary, known as rosemary oil, is extracted by hydrodistillation method from the fresh leaves and twigs. The yield ranges from 0.5 to 1.5% [w/w]. Improving the total yields, determining minimum steam to be passed through the packed bed, evaluating influence of used multi-stage column and trend to change into important component with time of distillation are the aims of this work. In this work, essential oil was extracted from rosemary plant by steam distillation method. For the extraction of essential oil by this method rosemary was exposed to the steam flow. The processes were done and the extraction yield was calculated. Then, the components of the essential oils analyzed by GC-MASS and GC instruments. Experiments that were conducted in multi-stage columns and decreased steam flow rate proved that an increase in the number of stages has a considerable effect upon the yield. Also analyses which were done on essential oils components during 5, 15, 30, 60 and 100 minutes intervals showed different extraction phenomena for three major components of the oil; a-pinene, 1, 8- cineol and camphor. This study showed a direct correlation between increase of contact time of steam and increase of the yield. Also barrier effect of cell wall and boiling point of components affect extraction phenomena of them

Study on seasonal variation of flavonoids in ginkgo biloba l. cultivated in Iran

Ginkgo is a native plant to China which is recently cultivated in Iran. The extract of the Ginkgo leaves contains flavonoids and ginkgolids and has been used pharmaceutically for a while. Flavonoids of Ginkgo have been shown to have several effects which could increase blood flow of brain and limit the damage associated with ischemia also they inhibit cyclo-oxygenases and lipoxygenases activities and show antioxidant effects. The researchers showed that Ginkgo leaves flavonol glycosides varies during different seasons. The aim of the present research is study on seasonal variation of flavonoids of Ginkgo biloba leave's cultivated in Iran by HPLC method. In this study Ginkgo leaves from 5 years old trees collected in 2005 from North of Iran and evaluated. Plant leaves were dried, milled, and extracted with a suitable solvent. For analysis of flavonoid aglycones in the extract, flavonol glycosides were hydrolyzed in acid medium and identified by HPLC instrument and the peaks of standard material; Quercetin, Kaempferol and Isorhamnetin were obtained. According to our findings, flavonoids vary in amount during seasons and upper most amount was found in June Month [5.5%w/w]. Results of our study shows a mild variation of flavonoids content in Ginkgo tree leaves during different months, also we found a high concentration of flavonoids in young tree leaves that is very higher than pharmacopeia's standard [0.5% w/w] and these leaves are suitable for preparing of standardized extracts