Phytopharmaceutical Preparations Research Articles

Antiproliferative activity of standardized herbal phytopreparation from Asclepias subulata.

Background: Several studies have shown that active compounds of Asclepias subulata (cardenolides) have antiproliferative effect on human cancer cells. Cardenolides isolated from A. subulata can be used as active chemical markers to elaborate phytopharmaceutical preparations. The aim of this work was to evaluate the antiproliferative effect of a standardized extract of the aerial parts, based on Asclepias subulata cardenolides. Methods: Four standardized extracts were prepared by HPLC-DAD depending on the concentration of calotropin and the antiproliferative activity was measured for the MTT assay, on the A549, MCF-7, HeLa, PC3 and ARPE cell lines. The concentrations of calotropin used for the standardization of the extracts were 10, 7.6, 5 and 1 mg/dL. Results: Standardization of the A. subulata extract based on calotropin at 7.6 mg/g dry weight was achieved and the antiproliferative activity was evaluated over A549, HeLa and MCF-7 cell lines, obtaining proliferation percentages of 3.8 to 13.4% . Conclusions: The standardized extracts of A. subulata at different concentrations of calotropin showed antiproliferative activity against all the cell lines evaluated. The greatest effect was observed against the HeLa cell line.

Antiproliferative activity of standardized herbal phytopreparation from Asclepias subulata.

Background: Several studies have shown that active compounds of Asclepias subulata (cardenolides) have antiproliferative effect on human cancer cells. Cardenolides isolated from A. subulata can be used as active chemical markers to elaborate phytopharmaceutical preparations. To evaluate the antiproliferative effect of a standardized extract of the aerial parts, based on Asclepias subulata cardenolides. Methods: Four standardized extracts were prepared by HPLC-DAD depending on the concentration of calotropin and the antiproliferative activity was measured for the MTT assay, on the A549, MCF-7, HeLa, PC3 and ARPE cell lines. The concentrations of calotropin used for the standardization of the extracts were 10, 7.6, 5 and 1 mg/dL. Results: Standardization of the A. subulata extract based on calotropin at 7.6 mg/g dry weight was achieved and the antiproliferative activity was evaluated over A549, HeLa and MCF-7 cell lines, obtaining proliferation percentages of 3.8 to 13.4% . Conclusions: The standardized extracts of A. subulata at different concentrations of calotropin showed antiproliferative activity against all the cell lines evaluated. The greatest effect was observed against the HeLa cell line.

Influence of Biopesticides and Natural Preparations on the Regulation of Diseases and Pests in the Ecological Protection of Tomatoes (Lycopersicon esculentum L.) and Cucumbers (Cucumis sativus L.)

Organic agriculture is increasingly present on farms in response to environmental pollution and food safety. However, organic farmers have a big production risk due to frequent plant diseases. This research was conducted to compare the influence of preparations for plant protection allowed in organic agriculture and phytopharmaceutical preparations which are used in conventional agriculture and observe their effect in regulation of pests and diseases in tomato and cucumber production in an open field. The research included the tomato variety „Oxheart“, and the cucumber variety „Long green“. The time of research was four months from planting to harvest (period: May 2020. – August 2020.) until more than 70% of vegetation was completed. The tests were performed at a location in the rural part of the town of Živinice (Tupković) in an open field with an altitude of 250 m, during the vegetation period of 2020. The tests were performed on two separate plots. The plot where phytopharmaceutical (conventional) preparations were used, was physically separated from the plot where preparations allowed in organic agriculture were used. Temperature measurement was performed using a TFA Dostmann thermometer (-25ºC to + 50ºC) and it was set at a height of 2 meters in the open field. The amount of rainfall was monitored with the TFA Dostmann rain gauge. In the tests with organic plant protection, the nettle solution and preparation with active substance Bacillus thuringiensis for pests was used, and for plant diseases, the milk+soda solution and copper-oxide were used. In the tests with conventional protection, preparations with active substances acetamiprid and imidacloprid for pests were used, and the diseases were treated with mancozeb+copper and metalaxyl-M+mancozeb. After treatments, the onset of diseases was monitored. To conclude, the conventional insecticides gave better results in the regulation of pests in comparison to organic preparations, and in the case of organic preparations, the nettle solution had a better effect. Plant protection with copper-oxide gave the same results as using conventional preparations in disease regulation. This shows that plant protection is possible in organic production, but preparations must be applied more often and in a timely manner.

Analysis of bioactive compounds and antioxidant activities of cultivated garlic (allium sativum l.) and red onion (allium cepa l.) in algeria

In all times, humankind has used several species of the genus Allium as food, spice, or herbal remedy. Some of these species have been cultivated, such as garlic (Allium sativum) or onion (Allium cepa). Today, their value for human health care is one of the most important aims of research. Up to now, many applications of Allium species are known for the use of phyto-pharmaceutical preparations. Therefore, the present study aimed to determine the phytochemical profile of cultivated garlic (Allium sativum), and red onion (Allium cepa) in Algeria, both quantitatively (total phenolic, total flavonoids, condensed and hydrolysable tannins contents) and qualitatively (phytochemical screening), to characterize the phenolic compounds using HPLC method and to evaluate the antioxidant properties using DPPH assay. Red onion gave the higher amounts of total phenolic compounds (86±1.00mg GAE/100g DM), flavonoids (43.33±0.57mg QE/100 g DM), condensed tannins (4.4±0.52 mg CE/100g DM) and hydrolyzable tannins (0.22±0.04mg TAE/100g DM) compared to garlic (45±1.00mg GAE/100g DM, 34.66±0.57mg QE/100g DM, 6.8±0.34mg CE/100g DM and 0.05±0.01mg TAE/100g DM) respectively. Five compounds were found in red onion extract and one compound in garlic extract after chromatographic analysis of the samples. Furthermore, red onion possessed the higher antioxidant activity (IC50= 420.9±5.00 µg/ml) as compared to garlic (919.87±4.43 µg/ml). These findings provide ample evidence of the existence of bioactive compounds in garlic and red onion, both of which are rich in phenolics primarily flavonoids and tannins, have strong antioxidant activity, and can be further consumed directly or as food products.

Application of sage herbal dust essential oils and supercritical fluid extract for the growth control of Escherichia coli in minced pork during storage

Sage has various biological activity and long tradition of the using in phytopharmaceutical preparations and, recently, in food industry. In this work, the essential oil obtained by hydrodistillation and essential oil and extract obtained by supercritical fluid extraction from sage herbal dust, waste product of filter tea factory, were screened for antimicrobial activity against Escherichia coli in the medium which simulates the conditions in meat and in real system of minced pork. The supercritical fluid extract had much lower effect to the reduction of a number of E. coli in meat simulation medium. The total inhibition of E. coli growth was achieved after 12 h and 24 h with the addition of 0.6 μl/ml of essential oil obtained by hydrodistilation and supercritical fluid extraction, respectively. The addition of essential oils and supercritical fluid extract of sage herbal dust could be used for the effective control of E. coli growth in minced meat.

The Potential Gastrointestinal Health Benefits of Thymus Vulgaris Essential Oil: A Review

In recent times, medicinal plants have received great attention worldwide due to their effective pharmacological properties and therapeutic benefits. Numerous chemical compounds extracted from various medicinal plants have manifold biological activities. Thymus vulgaris (TV) is a flowering plant with an aromatic odor that has been broadly applied in conventional medicine, food additives, and phyto-pharmaceutical preparations. It is recognized to have promising therapeutic potential for curing various types of diseases. The essential oil extracted from TV, which contains a high quantity of flavonoids, possess antioxidant and antimicrobial activities. Consequently, it could be utilized as a good source in developing novel natural antioxidants and antibiotics. This review explores some of the potential health benefits of TV essential oil (TVEO) on the gastrointestinal tract.

Phytochemical content, extract standardization and antioxidant activity of Calliandra calothyrsus Meissn leaf, a potential phytoestrogen source

Calliandra (Calliandra calothyrsus Meissn.) is a leguminous shrubs that almost spread throughout the area of Bali island, especially the area of Baturiti, Tabanan. The ethanol extract of 80% of C. calothyrsus leaves is a potential source of exogenous estrogen in terms of its phytoestrogen content. Potential extract of potassium which is very useful can be consumed in the form of phytopharmaca preparations. One of the requirements to prepare the formulation of phytopharmaceutical preparations should be done related to phytochemical screening and standardization of raw materials. This study aims to perform phytochemical screening and standardization of 80% ethanolic extract of C. calothyrsus leaves. Phytochemical screening can provide an overview of chemical compounds which are contained in C. calothyrsus leaf extracts to their activity. The standardization of extracts will ensure the quality of raw materials extract to be used. The leaf extraction of 80% ethanol yielded as much as 21.85%. Phytochemical screening showed that 80% positive ethanol extracts contained flavonoid group compounds. The powder standardization obtained 8.35% of water content. While the extracts standardization obtained 5.26% of extract water content. The total ash content of the extract was obtained 4.54% and the acid soluble ash content was 0.5%. The ethanol extract of 80% C. calothyrsus leaf has met the standardization of raw material extract.

Tetraglochin andina Ciald.: A medicinal plant from the Argentinean highlands with potential use in vaginal candidiasis

Ethnopharmacological relevanceThe Argentinean medicinal plant Tetraglochin andina Ciald, formerly classified as T. cristatum (Britton) Rothm is used in traditional medicine by inhabitants from Argentinean northwestern highlands (Puna) to treat candidiasis and as anti-inflammatory. Aim of the studyTo assess the potential of the crude drug as an anti-Candida agent with anti-inflammatory properties. The bioactivity and phytochemical composition of a dry extract of the plant was investigated. Material and methodsThe pharmacognostic description of the crude drug is carried out for the first time, including macroscopic and microscopic examinations of the different organs, physicochemical and extractive values (petroleum ether-, ethanol- and water-soluble). The dry extract from T. andina was evaluated as antifungal against pathogenic Candida sp. and Saccharomyces cerevisiae isolated from vaginal infections and reference strains, by the macrodilution and microdilution assays. The normal vaginal microbiome in women is characterized by the dominance of lactic acid-producing bacteria, mainly Lactobacillus spp. The effect of T. andina extract on Lactobacillus strains was also assayed. The inhibitory effect on proinflammatory enzymes (cyclooxygenase, lipoxygenase and phospholipase A2) and antioxidant capacity was studied. The chemical profile was analyzed by HPLC-ESI-MS. ResultsThe hydroalcoholic extract inhibited the growth of all yeasts with Minimal Inhibitory Concentration (MIC) values between 12.5 and 400 µg GAE/mL and the MIC values on Lactobacillus were higher than the MIC values against Candida isolates ( > 400 µg GAE/mL). These results indicate that the hydroalcoholic extract could be used without affecting the normal microbiota of vaginal fluid. The extract showed antioxidant activity and could modulate the inflammatory process by three pathways (sPLA2, COX-2, LOX). The plant extract contained high total phenolic levels (386.9±1.7 mg GAE/g dry extract) and flavonoid levels (260.4±2.7 mg GAE/g dry extract). Fifty phenolic compounds were identified by HPLC-ESI-MS. They were mainly hydrolysable and condensed tannins. The dry extract was chemically and biologically stable during one year at room temperature or 4 °C. ConclusionsThe presence of anti-Candida and anti-inflammatory activities in Tetraglochin andina extracts give support to their traditional use for treating conditions associated with microorganism infections and inflammatory process in humans. This plant preparation could be used to design phytopharmaceutical preparations to inhibit yeast growth and moderate the inflammatory and oxidative process.

Evaluation of Asteraceae herbal extracts in the management of diabetes and obesity. Contribution of caffeoylquinic acids on the inhibition of digestive enzymes activity and formation of advanced glycation end-products (in vitro)

The study was performed to assess, for the first time, the in vitro anti-diabetic potential of ten Asteraceae plant extracts to inhibit the activity of digestive enzymes (α-amylase, α-, β-glucosidases and lipase) responsible for hydrolysis/digestion of sugar and lipids. Prevention of advanced glycation end-products (AGEs) formation was evaluated in bovine serum albumin/ribose glycation reaction model. The phytochemical profiles and caffeoylquinic acids (CQAs) contents were determined for the methanolic extract of each plant. Analyzed plant extracts exhibited significant inhibitory activity against key digestive enzymes linked to type II diabetes and obesity. A strong inhibition was observed for glucosidases and mild activity towards amylase and lipase (compared to reference compounds). Moreover, some extracts exhibited potent ability to prevent formation of AGEs, implicated in some diabetic complications. Caffeoylquinic acids were dominant in all plant extracts and findings demonstrate that these compounds are the most relevant hypoglycemic and anti-glycation agents. From the obtained results, Argyranthemum pinnatifidum, Helichrysum melaleucum, and Phagnalon lowei are good candidates for further development of phyto-pharmaceutical preparations as complementary therapy for diabetes and obesity control.

The effects of supplemented diets with a phytopharmaceutical preparation from herbal and macroalgal origin on disease resistance in rainbow trout against Piscirickettsia salmonis

The present study aimed to evaluate the effects of a commercial phytopharmaceutical preparation from herbal and macroalgal origin on the growth and immune response of rainbow trout adapted to seawater and its susceptibility to Piscirickettsia salmonis infection. Preliminary in vitro trials, evaluated the effects of the commercial product Futerpenol® on the expression levels of selected immune-regulatory genes and its protective effect in a challenge against Piscirickettsia salmonis (LF89). Subsequent in vivo feeding trials were conducted to corroborate fish protection against Piscirickettsia salmonis. Control and treatment diets (with or without the commercial product Futerpenol® at a concentration of 1kg/ton) were fed to triplicate groups of 50 fish (average weight: 100.1±11.1g) during 30days. Fish from all dietary groups were equally redistributed in three tanks and challenged by cohabitation with fish infected with P. salmonis (shedders) at the end of the feeding treatment, and mortalities were recorded over 80days post-infection. A cumulative mortality of 35.0±4.3 and 15.0±6.0% was registered when challenged fish were previously fed during 30days with the control and treatment diets respectively. Futerpenol® dietary administration preceding the cohabitation challenge gave significant protection from P. salmonis as suggested by the Relative Percent Survival (RPS) values of 62.3 and 57.1% after 60 and 80days post-infection, respectively. These results suggest that dietary supplementation with Futerpenol® at 1kg/ton do not affect growth performance and strengthen immunity of Oncorhynchus mykiss against P. salmonis under the experimental conditions applied during this study. Statement of relevanceThis study demonstrates that dietary supplementation with a phytopharmaceutical preparation, rich in fucoidans and labdane diterpenes strengthen immunity of O. mykiss against P. salmonis. The use of feed supplemented with this kind of phytopharmaceutical preparations in anticipation to potential infections or vulnerable stages of the production could be useful as an alternative method to prevent and limit the outbreak of salmonid rickettsial septicaemia (SRS) which constitutes one of the main infectious diseases causing production problems and substantial loss in salmonids and marine fish aquaculture worldwide.

Quantification of methyl esters of fatty acids in the oil of Physalis minima by GC-MS

Objective: To investigate quantification of methyl esters of fatty acids in the oil extracted from Physalis minima (P. minima) using gas chromatography-mass spectrometer. Methods: Oil was extracted from the shade dried plant with n-hexane through Soxhlet extraction. Fatty acids that present in the oil were derivatized to fatty acid methyl esters and analysed through gas chromatography-mass spectrometer. Results: A total of nine fatty acids were detected in quantifiable amount in the oil. Both the saturated fatty acids and unsaturated fatty acids were identified. Palmitic acid was found in the highest concentration as 46.83%. Linoleic acid (ω-6) and linolenic acid (ω-3) were obtained in appreciable amount as 16.98% and 14.80% respectively among the unsaturated fatty acids in the oil under study. From the literature review, it appeared that fatty acids were determined for the first time in the oil of P. minima. Conclusions: Presence of these important fatty acids in high amount makes P. minima oil beneficial for health, which can be used in the preparation of phytopharmaceutical or pharmaceutical preparations. Moreover, the results of this study are useful for the phytopharmaceutical industries to establish their quality control profile.

Hypericins: biotechnological production from cell and organ cultures.

Hypericum perforatum L. (St. John's wort), a perennial flowering plant native to Europe, is widely used as a medicinal plant and has a long history of its use in the treatment of various ailments. Currently, H. perforatum is widely used as an herbal remedy for the treatment of mild to moderate depression. Hypericins are natural napthodianthrone compounds produced from H. perforatum (St. John's wort) which are having antitumor, antiviral (i.e., against human immunodeficiency and hepatitis C virus), antineoplastic, and antidepressant properties. Currently, field-grown plant materials are generally used for the commercial production of hypericins. It has been reported that hypericin accumulation in natural plants is influenced by different ecological and environmental conditions including light intensity, nitrogen availability, temperature, seasons, and growing regions. Therefore, up to 17-fold and 13-fold differences in hypericin and pseudohypericin amounts, respectively, are reported in different phytopharmaceutical preparations. Plant cell and organ cultures are effective systems for producing natural products, and attempts were made for the production of biomass and stable concentrations of hypericins through in vitro cultures of H. perforatum. Cell, callus, shoot, plantlet, and adventitious root cultures have been established and various chemical and physical factors which influence the biomass and secondary metabolite accumulation have been investigated. Large-scale plantlet and adventitious root cultures have also been attempted in H. perforatum in bioreactors, and various strategies have been applied for the production of higher biomass and secondary products. This review describes the biotechnological approaches employed for the production of hypericins and focuses upon the challenges and future prospects.

Assessment of stability of a spray dried extract from the medicinal plant Bidens pilosa L.

Spray drying has been successfully employed for the encapsulation of herbal bioactive compounds resulting in stable phytopharmaceutical preparations. Bidens pilosa L. is a South American medicinal plant with proved antimalaric, hepatoprotector and antioxidant activities, generally linked to their secondary metabolites, flavonoids and polyacetylenes. In this work the physicochemical stability of an optimized spray dried composition from a B. pilosa extract was evaluated at three different stress storage conditions in open containers and in sealed sachets. High performance liquid chromatography was employed to monitor the concentration of three marker compounds over 12months. Color variation of the stored samples was evaluated by using a color spectrophotometer. It was observed that the concentration of the monitored compounds of the plant decreases more drastically in samples stored in open containers. The two flavonoids monitored, rutin and hyperoside, showed lower degradation than the polyacetylene. The concentration of the markers did not change significantly at the lowest temperature. With regard to color, darker hues were observed at higher temperatures and storage times. This study showed that the storage conditions cause significant impact on stability of standardized spray dried B. pilosa extract.

Morphological and chemical characterization of Arnica montana L. under different cultivation models

Arnica (Arnica montana L.) is a perennial mountain plant whose flower heads are mainly used for topical treatment of bruises and sprains in phytopharmaceutical preparations such as ointments, creams and gels. In this study, different models of arnica cultivation have been tested in three-year field trial arranged in 2×3×2 factorial design. Planting date (spring and autumn), fertilizater type (manure, NPK and control) and mode of reproduction (generative and clonal) were induced factors on the trial. The variation of the rosette diameter, flowering stem count and height, flower heads count and diameter, together with flower heads yield and chemical properties were investigated as morphological parameters. Spring planting of clonally propagated plants was showed to be the least successful model due to the low percent of rooted plants and weak shoot development. Other models yielded from 59.8 to 143.6kgha−1 and 116.3 to 258.7kgha−1 flower heads (dw) in the second and third year, respectively. Total sesquiterpene lactones content in flower heads of all tested variants was above the minimum of pharmacopeia quality regulation and ranged from 4.6 to 13.9mgg−1 depending on factor levels, while the amounts of two dominant flavonoid compounds quercetin-3-O-glucoside and kaempferol-3-O-glucoside ranged from 7.8 to 13.9mgg−1 and from 2.1 to 4.7mgg−1, respectively. All induced factors in the field trial significantly influenced variation in both morphological and chemical parameters, where fertilized variants from autumn planting have shown to be superior in flower heads yield with satisfying levels of the secondary metabolites content. Factorial analysis distinguished that half of morphological parameters positively correlated with flower heads yield. First three ranks of sequential classification of cultivation models with desirable properties were captured by variants planted in autumn, while the last rated variants were those from clonal propagation planted in spring. This attempt of arnica domestication in agroecological conditions of western Serbia has proven to be successful in ensuring high quality raw material, providing at the same time answers to the questions that have not been reviewed so far.

Antimalarial activity of two spray dried formulations of Bildens pilosa L. based on Brazilian ethnopharmacology

Bidens pilosa L. (Asterarecae), a plant native of South America is employed in the Amazon region, for the treatment of malaria. This biological activity, seem to be associated to the presence of polyacetylenes and phenolic compounds as flavonoids (Brandão et al., 1997). According to World Health Organization, each year, occurs 219 million cases of malaria in the world resulting in about 660.000 deaths, mostly in children of low income countries (WHO, 2012). Medicines based on phytopharmaceutical preparations for treatment of endemic diseases such as malaria are interesting strategies justifying the research in this area. The spray drying technique have been successfully applied for the production of solid delivery systems containing plant extracts. The aim of this work was to evaluate the in vivo antimalarial activity of two spray dried formulations prepared with the hydroethanolic extract of roots of B. pilosa employing β-cyclodextrin and Aerosil:Microcrystalline celulose (MC) as carriers. Antimalarial activity was performed in mice infected with Plasmodium berghei, NK-65 by an approved ethical protocol. The presence of flavonoids and polyacetylens was monitored by high performance liquid chromatography. Results showed that the formulation containing Aerosil®:MC increased in 75% the mice survival, at a concentration of 150 mg/kg, the formulation containing β-cyclodextrin also increased the mice survival at all the doses tested being the concentration of 50 mg/kg the most effective with a 50% survival. Both formulations reduce parasitemia the 5th day compared with the control group. The preparations developed can be used as adjuvants for the treatment of malaria.

Bioactive compounds in Bidens pilosa L. populations: a key step in the standardization of phytopharmaceutical preparations

The total flavonoid content (TFC), total polyphenols content (TPC), and in vitro antioxidant activity (AA) of six Bidens pilosa L., Asteraceae, populations harvested from different localities were evaluated in this work. The plants were separated in roots, stems, and leaves/flowers, and the influence of extraction methods was investigated. A reversed-phase high-performance liquid chromatography method (HPLC) was developed and employed to obtain characteristic HPLC fingerprints of the bioactive compounds present in the extractive solutions, which were correlated with the TFC, TPC, and AA. Extractive solutions of leaves/flowers presented a higher AA when compared with those obtained from other parts of the plant (IC50 of 35.35±0.10μg/mL). The stem extracts presented the lowest AA (IC50 117.2±1.96μg/mL). A direct correlation of AA with TFC and TPC was evidenced. The highest AA was obtained by dynamic maceration and was statistically different from the AA presented by the extractive solutions obtained by other extraction methods. The results of this work evidenced differences that can be found at different stages of development of phytopharmaceutical preparations from B. pilosa and highlighted the importance of using the concentration of marker compounds as well as HPLC fingerprints as quality control parameters.

Physicochemical Properties of Phytopharmaceutical Preparations as Affected by Drying Methods and Carriers

This article reports a study of the effects of the drying methods and drying carriers on system performance and physicochemical properties of spray- and spouted bed–dried phytopharmaceutical preparations from Bidens pilosa L. Colloidal silicon dioxide, β-cyclodextrin, maltodextrin dextrose equivalent (DE) 10, and microcrystalline cellulose were used as drying carriers. The dried product was characterized by the particle size and morphology, total flavonoid content, solubility, flowability, and water activity. High-performance liquid chromatography (HPLC) was used to detect four marker compounds previously reported for this plant. Spray and spouted bed drying systems were compared through energetic efficiency, product recovery, elutriation, and product accumulation. The crystalline state of the powders was assessed by X-ray diffraction. Results showed high degradation rates for total flavonoid content and marker compounds during spouted bed drying. Depending on the drying carrier added to extractive solutions, different degrees of protection from degradation were conferred, and the physical characteristics of the product were changed accordingly. The lowest flavonoid degradation (8.6%) and the higher concentration of marker compounds were obtained using β-cyclodextrin as the drying carrier. Particle size was higher when a blend of Aerosil (Evonik Degussa, Hanau, Germany) and cellulose was employed as the drying carrier. A maximum product recovery of 86.9% was achieved by spray drying and 72.9% by spouted bed drying with the Aerosil : cellulose composition.

Rhizome and root yield of the cultivated Arnica montana L., chemical composition and histochemical localization of essential oil

Arnica montana (L.) is a medicinal plant whose flower heads are widely used for the topical treatment of bruises and sprains in phytopharmaceutical preparations. Despite being cultivated for flower heads production for a long time, the quantity and quality of its underground parts have not received much attention. In this study underground parts of a three-year cultivation trial have been estimated for yield potential and chemical composition of essential oil in two consecutive years. Yields of rhizome dry mass were about 208kgha−1 and 647kgha−1, and root dry mass were about 330kgha−1 and 525kgha−1 in the second and third year of growing period, respectively. The chemical composition of the essential oil from root and rhizome, obtained by hydrodistillation, was analyzed by gas chromatography–flame ionization detection and GC–mass spectrometry. At the end of second year essential oils from rhizome and root, yielded on average about 4.05% and 1.89%, and at the end of third year, they yielded 2.43% and 1.85%, respectively. The composition of essential oils was independent regarding cultivation trial factors with small differences regarding years. Aromatic hydrocarbons were predominant group of essential oil constituents (about 84% in average) and were identified as thymol derivates. Main individual components were 2,5-dimethoxy-p-cymene, thymol methyl ether and 2,6-diisopropylanisole. Furthermore, histochemical localization of essential oils in rhizome and root was carried out. Secretory structures responsible for essential oil synthesis and accumulation were studied using light, scanning and transmission microscopy. Secretory tissue is localized in the cortical region of the root and rhizome in form of idioblastic secretory cells and secretory cavities, respectively. Histochemical analyses confirmed the lipid nature of secreted material.

Effects of Thymol and Carvacrol, Constituents ofThymus vulgarisL. Essential Oil, on the Inflammatory Response

Thyme (Thymus vulgaris L., Lamiaceae) is an aromatic and medicinal plant that has been used in folk medicine, phytopharmaceutical preparations, food preservatives, and as an aromatic ingredient. The effect of Thymus vulgaris essential oil (TEO) and its isolated constituents thymol and cavacrol (CVL) were studied in the following experimental models: ear edema, carrageenan-induced pleurisy, and chemotaxis in vitro. In the pleurisy model, TEO, CVL, and thymol significantly inhibited inflammatory edema. However, only TEO and CVL inhibited leukocyte migration. In the in vitro chemotaxis experiment, CVL inhibited leukocyte migration, whereas thymol exerted a potent chemoattractant effect. In the ear edema model, CVL (10 mg/ear), applied topically, reduced edema formation, exerting a topical anti-inflammatory effect. Thymol did not reduce edema formation but rather presented an irritative response, probably dependent on histamine and prostanoid release. Our data suggest that the antiinflammatory effects of TEO and CVL are attributable to the inhibition of inflammatory edema and leukocyte migration.

Investigação da presença de anorexígenos, benzodiazepínicos e antidepressivos em formulações fitoterápicas emagrecedoras

Several weight-reducer phytotherapic formulations commercialized in Brazil contain undeclared synthetic drugs. This situation constitutes a public health issue for the country. This study aimed at evaluating the occurrence of anorexigenic, benzodiazepinic and antidepressant drugs in the phytopharmaceutical preparations monitored and collected by the Public Health Surveillance of the State of Minas Gerais, Brazil. By chemical tests, the absence of at least one class of the analyzed secondary metabolites (flavonoids, saponins and quinones) was found in all analyzed samples. Fenproporex, fluoxetine and chlordiazepoxide, alone or in association, were identified by GC-MS in all formulations. Therefore, the analyzed samples did not comply with the established quality standards, as these formulations contain undeclared synthetic compounds in their compositions, along with the association of anorexigens and anxiolytics drugs, which are not allowed by the Brazilian sanitary legislation. Hence, a more effective quality control of the weight-reducer phytotherapy formulations is demanded, as well as improving educational actions addressed to health professionals and consumers.