Extract Of Artemisia Annua Research Articles

Validation, transfer and measurement uncertainty estimation of an HPLC–UV method for the quantification of artemisinin in hydro alcoholic extracts of Artemisia annua L.

Malaria is the world's most important parasitic infection with 500 millions cases annually and almost 2 millions death per year. This disease is more present in Sub-Saharan Africa where 90% of the infections are found. Artemisinin and its semi synthetic derivatives (artemether, artesunate) have actually the most powerful activity on malaria, even in its complicated forms and resistance cases. Various methods have been proposed for detection and quantification of artemisinin in Artemisia annua L. by HPLC–UV, but the plant extracts used for this quantification were extracts obtained with organic solvents (toluene, petroleum ether, hexane). To be able to use crude A. annua extracts prepared at low cost to formulate antipaludic drugs, we chose the use of a mixture of water and ethanol as solvent of extraction, but no adequate analytical method for this kind of extracts is published. The main objectives of this work were first to develop an analytical method for artemisinin quantification in hydro alcoholic extracts of A. annua. Second, this method had to be thoroughly validated by the research and development laboratory and, third, the transfer of this method to the routine laboratory had to be demonstrated. The final aim was to compare the estimation of measurement uncertainty obtained during the method validation with validation standards to measurement uncertainty estimates obtained during the method transfer study with real samples. The method was validated following the accuracy profile methodology and was found to be accurate in the concentration range of 10.0–54.0 μg/ml with CV < 8%. Limit of detection and of quantification were 2.73 and 10.0 μg/ml, respectively. The method was then successfully transferred to a laboratory in Benin by showing that the quality of the results that it will generate during routine application of the method is sufficient. Finally, the measurement uncertainty of the method was estimated from the validation experiments as well as from the transfer study with authentic unspiked samples of A. annua. The comparison of these measurement uncertainty estimations showed that they were coherent. It confirmed thus that the estimation of measurement uncertainty from validation experiments predicts well the measurement uncertainty of real routine samples. This analytical method was thus shown to be convenient for routine analysis of hydro alcoholic extracts of A. annua in Benin.

Status of carbohydrate, protein and lipid profile in the mosquito larvae treated with certain phytoextracts

Objective To investigate the impact of the most potent petroleum ether extract of Artemisia annua (A. annua) and Azadirachta indica (Az. indica) on total carbohydrate, lipid and protein level of Anopheles stephensi and Culex quinquefasciatus larvae. Methods Mosquito larvae were exposed to the extracts selected as per standard WHO procedure. Carbohydrate (glucose), total lipid and protein were estimated by the methods as Nelson, Bragdon and Lowry described, respectively. Results The glucose levels were increased to 27.87% and 46.8%, respectively in anopheline larval tissues after treatment with petroleum ether extract of A. annua and methanolic extract of Az. indica. In culicine larvae, glucose levels were reduced to 58.96% and 24.65%, respectively. After treatment with A. annua extract, lipid contents in anopheline and culicine larvae decreased by 28.57% and 25.0%, respectively and increased by 14.29% and 50.00% in the Anopheles and Culex larvae, respectively after treatment with methanolic extract of Az. indica. Total protein levels were reduced to 63.13% and 92.62% in anopheline and to 32.39% and 48.12% in culicine larvae after treatment with A. annua and Az. Indica extracts, respectively. Conclusions Two extracts produce significant alterations in the biochemical profiles of anopheline and culicine larvae. Further, the impacting factors of extracts on carbohydrate, lipid and protein contents of larvae are species and specific extraction. It indicates the disturbed metabolic activity of the larvae.

Artemisia annua extracts, artemisinin and 1,8-cineole, prevent fruit infestation by a major, cosmopolitan pest of apples

Context: Extracts of Artemisia annua (L.) (Asteraceae) and artemisinins are used for treatment of malaria, parasitic infections and have potent anticancer properties in cell lines. Eucalyptus oil and 1,8-cineole have antimicrobial, immune-stimulatory, anti-inflammatory, antioxidant, analgesic, and spasmolytic effects. Codling moth, Cydia pomonella, (L.) (Tortricidae), is a major cosmopolitan pest of the apple, potentially causing damage translating to 40 billion US dollars per year, globally. Currently used control measures are either hazardous to agricultural workers and harmful to environment, or ineffective. The potential of plant-derived semiochemicals for codling moth control is heavily understudied.Objective: This study evaluated the potential of A. annua extracts, and two chemicals that this plant contains, artemisinin and 1,8-cineole, for preventing apple feeding and infestation by neonate Cydia pomonella larvae.Methods: We studied effects of A. annua extracts, artemisinin and 1,8-cineole on apple infestation by neonate codling moth larvae using fruit choice assay in laboratory experiments. Preference of fruit treated with test solutions versus fruit treated with solvent was recorded and analyzed.Results: Crude A. annua extracts prevented fruit feeding at 1, 3, and 10 mg/ml. Artemisinin had feeding deterrent effects at 10 and 30 mg/ml, and 1,8-cineole at 100 and 300 mg/ml.Discussion and Conclusions: A. annua contains chemicals that prevent apple infestation by codling moth neonates. Artemisinin and 1,8-cineole are among them, but there are other, polar constituents of A. annua, which have similar effects. There is a potential of using our findings in codling moth control and production of codling moth-resistant apples.

In vitro activity of Artemisia annua L (Asteraceae) extracts against Rhipicephalus (Boophilus) microplus

The activity of plant extracts on parasites may indicate groups of substances that are potentially useful for controlling Rhipicephalus (Boophilus) microplus. The aim of the present study was to investigate the in vitro action of Artemisia annua extracts on this tick. The concentrations of the sesquiterpene lactones artemisinin and deoxyartemisinin present in plant extracts were quantified via high-performance liquid chromatography. Four extracts produced from the concentrated crude extract (CCE) were evaluated on larvae using the impregnated paper method, with readings after 24 hours of incubation. The engorged females were immersed in the CCE and in its four derived extracts for five minutes, with incubation for subsequent analysis of biological parameters. The extracts were not effective on the larvae at the concentrations tested (3.1 to 50 mg.mL(-1)). The CCE showed greater efficacy on engorged females (EC(50) of 130.6 mg.mL(-1) and EC(90) of 302.9 mg.mL(-1)) than did the derived extracts. These results tend to confirm that the action of artemisinin on engorged females of R. (B.) microplus is conditional to their blood intake. In this case, in vitro methods would be inadequate for effective evaluation of the action of A. annua on R. (B.) microplus.

Analysis of artemisinin in plant extracts of Artemisia annua L. cultivated in Brandenburg

Analysis of artemisinin in plant extracts of Artemisia annua L. cultivated in Brandenburg

Chromatography‐Crystallization Hybrid Process for Artemisinin Purification from Artemisia annua

AbstractThe feasibility of using a chromatography‐crystallization hybrid separation process for isolation and purification of artemisinin from the herbal plant Artemisia annua was studied. After separating the crude extracts of Artemisia annua by flash column chromatography, a two‐step antisolvent crystallization process was used for further isolation and purification. The two‐step antisolvent crystallization process was designed based on the solubility of artemisinin in different solvents and solvent mixtures. The first step crystallization process was to remove impurities from the solution and the second step to isolate artemisinin. The crystallization behavior of the two polymorphs of artemisinin during the antisolvent crystallization process was also studied by performing crystallization in solutions of artemisinin in acetonitrile and acetone, respectively. It was observed that the formation of the two polymorphs of artemisinin in acetone solution follows the Ostwald's rule of stages. Fast feeding of the antisolvent (water) resulted in crystallization of the metastable triclinic form which then transformed to the stable orthorhombic form through a solvent‐mediated transformation mechanism. The stable orthorhombic form crystallized out when water was fed slowly. Crystallization from acetonitrile always yielded the stable orthorhombic form regardless of the feeding rate of the antisolvent. Finally, a chromatography‐crystallization hybrid separation process was proposed and tested. The results of the present work demonstrated the feasibility of combining the advantages of column chromatography and crystallization to improve efficiency of the isolation of artemisinin from Artemisia annua.

Effects of ethanol extract of Artemisia annua L. against monogenean parasites of Heterobranchus longifilis

Ethanol extract of Artemisia annua was effective in the dislodgement and mortality of monogenean parasites of juvenile Heterobranchus longifilis at concentrations ranging from 50 to 200 mg/l. Five hundred 1-week-old juvenile fish were stocked in hapa in earthen pond for 7 days to accumulate parasites. The approximate number of parasites per fish was confirmed by counting the number of parasites attached to body surfaces and the gills with a stereo-microscope before being exposed to the extract under in vivo conditions. The bioactivity of the extract was conducted in plastic Petri dishes with three replications and controls. The results obtained from A. annua extract were matched against those produced by pure artemisinin and artesunate powder, respectively, under similar experimental conditions. There was a faster effect of pure artemisinin crystals on the parasites as compared to A. annua extract and artesunate. Coagulation of parasite cells was observed with artemisinin treatment, whereas parasites were merely dislodged from their attachment organs and killed some hours later in the same concentration of A. annua. There were positive correlations between the number of parasites dislodged/killed and the concentration of A. annua extract, artemisinin, and artesunate powder, respectively, as well as the duration of exposure of affected fish to the substances. This led to the conclusion that A. annua contains substances that are effective against helminthes parasites of H. longifilis.

Metabolomic investigation of the ethnopharmacological use of Artemisia afra with NMR spectroscopy and multivariate data analysis

Ethnopharmacological relevance Artemisia afra has been used as an infusion to treat malaria throughout the southern parts of Africa, in much the same way as the antimalarial plant Artemisia annua in China. The antiplasmodial activity of purified components from an apolar fraction of Artemisia afra has been shown in the past. No data on the efficacy of the tea infusion prepared from Artemisia afra are however available. Objective To investigate the antiplasmodial activity of various extracts of Artemisia afra including an ethnopharmacological prepared sample. To identify polar metabolites in Artemisia afra and Artemisia annua and by using multivariate data analysis investigate the metabolic differences between these species. Materials and methods The antiplasmodial activity of Artemisia afra and Artemisia annua extracts were tested for activity against Plasmodiam falciparum 3D7 (chloroquine-sensitive strain) with chloroquine, quinine and artemisinin as positive controls. Hydrophilic metabolites in Artemisia afra and Artemisia annua were identified directly from the crude extracts through 1D- and 2D-NMR spectra. The NMR spectra were also used to differentiate between the two species using principal component analysis (PCA) for quality control purposes. Results The apolar fractions of both Artemisia afra and Artemisia annua showed activity against P. falciparum while activity was only found in the tea infusion of Artemisia annua. Metabolomic studies using 1D- and 2D-NMR spectroscopy identified 24 semi-polar components in Artemisia afra including three new phenylpropanoids for this species: caffeic acid, chlorogenic acid and 3,5-dicaffeoyl quinic acid. PCA analysis conducted on the samples yielded good separation between the polar extracts of Artemisia afra and Artemisia annua. Conclusion These findings shows that there are no in vitro activity in the tea infusion of Artemisia afra and lists the identified metabolites causing the metabolic differences between Artemisia afra and Artemisia annua for quality control purposes.

Immunosuppressive effect of ethanol extract of Artemisia annua on specific antibody and cellular responses of mice against ovalbumin

Artemisia annua has been widely used to treat autoimmune diseases such as systemic lupus ­erythematosus and rheumatoid arthritis in traditional Chinese medicine. In this study, the ethanol extract of A. annua (EEAA) was evaluated for the immunosuppressive potentials on mice splenocyte proliferation in vitro, and the specific antibody and cellular immune responses in the ovalbumin (OVA)-immunized mice. EEAA significantly suppressed concanavalin A (Con A)- and lipopolysaccharide (LPS)-stimulated splenocyte proliferation in vitro in a concentration-dependent manner. EEAA also significantly suppressed Con A-, LPS- and OVA-induced splenocyte proliferation in the OVA-immunized mice in a dose-dependent manner. Meanwhile, the OVA-specific serum IgG, IgG1 and IgG2b antibody levels in the OVA-immunized mice were markedly reduced by EEAA. The results suggest that EEAA could suppress the cellular and humoral response in mice. This study provided evidence to understand the therapeutic effects of A. annua for treatment of some autoimmune diseases and an immunosuppressive natural products to further researches to be developed as immunosuppressant.

Synthetic Biology in Pursuit of Inexpensive, Effective, Anti-Malarial Drugs

One of the big success stories in synthetic biology is the re-engineering of an existing organism to produce the anti-malarial drug artemisinin. The drug is also found naturally in extracts of the plant Artemisia annua (Wormwood), but demand is far oustripping supply and extracting the drug from the plants is complex, time-consuming and expensive. In this lecture, Jay Keasling describes how he and his laboratory re-designed yeast microbes into living mini anti-malaria drug factories producing artemisinin more cheaply and efficiently. The lecture was given at the 12 May 2009 launch of the Imperial-based Centre for Synthetic Biology and Innovation—established in partnership with the London School of Economics and funded by the Engineering and Physical Sciences Research Council (EPSRC) as part of a science and innovation award.

Antitrypanosomal and antileishmanial activities of organic and aqueous extracts of Artemisia annua

Antitrypanosomal and antileishmanial activities of organic and aqueous extracts of Artemisia annua

Development of HPLC analytical protocols for quantification of artemisinin in biomass and extracts

Quantification of artemisinin purity and amount in plant material and extracts to date has been characterized by a considerable inconsistency in values. This is likely to be due to the adoption of varied analytical procedures and use of inappropriate to the specific applications analytical techniques. In this paper we are attempting to further develop artemisinin analysis to the point where a universally acceptable reference method is available to the research and end-users communities. Thus, we have developed and validated an HPLC-RI method and optimized an HPLC-ELSD method. We used the gradient HPLC-UV method recommended by the current artemisinin monograph as a comparison for the method improvements presented herein, and show the limitations for its application scope. The data reported should help to allow more reliable laboratory analysis of artemisinin in both pure samples and in Artemisia annua extracts.

Simultaneous Nonchiral Determination of Artemisinin and Arteannuin B in Artemisia annua Using Circular Dichroism Detection

A new method was performed using on-line coupling of nonchiral reversed-phase liquid chromatography (RP-LC) to circular dichroism (CD) spectroscopy for simultaneous determination of artemisinin and arteannuin B in crude plant extracts of Artemisia annua. Analysis was carried out on an LC–CD system equipped with an Agilent TC-C18 column (4.6 mm I.D. × 200 mm L, 5 μm) using gradient of acetonitrile. The method was validated to be practicable and reliable at alterable wavelength in the ranges of 220–420 nm as desired. LOD and LOQ of artemisinin and arteannuin B were 0.08, 0.26 and 0.31, 1.02 μg mL−1, respectively. It was more sensitive than conventional LC–UV and comparatively cheaper than LC–MS in analysis of TCM.

Hemin Modified Carbon Fat Electrode for Analyzing Antimalarial Endoperoxide Artemisinin in Artemisia annua

AbstractA hemin bulk modified carbon electrode with Adeps neutralis (solid fat) as binder was developed for the determination of antimalarial endoperoxide artemisinin in plant matrix. The hemin modified electrode showed significant catalytic activity for the electrochemical reduction of artemisinin at about −380 mV vs. Ag/AgCl in phosphate buffer solution of pH 7 by using cyclic and differential pulse voltammetry. Under optimized conditions strict linearity between artemisinin concentration and height of the cathodic catalytic current peak was observed in 4.8×10−6–7.8×10−5 M concentration range (R=0.9991) when using differential pulse voltammetry. The detection limit was calculated as 1.4×10−6 M of artemisinin. The developed electroanalytical device is suitable for the determination of artemisinin in Artemisia annua extracts.

Effects of Artemisia annua L. (Asteracea) on nutritional physiology and enzyme activities of elm leaf beetle, Xanthogaleruca luteola Mull. (Coleoptera: Chrysomellidae)

Methanolic extract of Artemisia annua L., a weed collected around paddy fields in north of Iran, was investigated for its toxic effects on: feeding, growth, fecundity, fertility including the biochemical characteristics of elm leaf beetle Xanthogaleruca luteola Mull. The toxicity tests indicate that insecticidal effect was 2.4-fold higher in 3rd instar larvae than in adults. The LC 50 was 48% and 43.77% of a methanolic leaf extract at 24 and 48 h, respectively. The feeding deterrence was highest at 10% concentration and lowest at 0.625% for 3rd instar larvae and adults during the first 24 h and decreased thereafter. Larval duration significantly increased at higher concentrations. Larval treatment with 10% methanolic extract resulted in deformed adults (adultoids) with drastic changes in gross morphology of female reproductive system. Food digestion and assimilation significantly decreased in adults. Larval duration and adult fecundity and fertility were also significantly reduced. Twenty-four hours after treating 3rd instar larva with the extract the levels of glucose, protein, urea, uric acid, α-amylase, alkaline phosphatase, alanine amino transferase and aspartate amino transferase significantly changed. However, at 48 h the extract lost its potency. The extract did not effect the level of cholesterol in treated larva. These results indicate that Artemisia annua L. extract contains inhibitors of key metabolic pathways that may be useful in future control of the elm leaf beetle.

Growth Inhibitory Nature of Artemisia annua Extract against Culex quinquefasciatus (Say)

Petroleum ether (Pee), carbon tetrachloride (Cte) and methanol extract (Mee) of Artemisia annua, Chenopodium album and Sonchus oleraceus were screened for their efficacy against Culex quinquefasciatus larvae. Pee of A. annua, Mee of A. annua and Ch. album, Cte of A. Annua were found effective in descending order after 24 and 48 hrs of treatment. Pee of A. annua, the most potent extract with LC 50 78.2 ppm was selected to study its influence on the development and metamorphosis of the culicine mosquito. The extract significantly affected the hatching, larval development, pupal transformation and also lengthened the larval and pupal periods. Growth index was remarkably reduced. Treated culicine eggs, larvae and pupae showed deformities including disruption of the body wall, distorted alimentary canal, damaged tracheal network and arrested histogenesis. The extract has remarkable effect on the metamorphosis and high larvicidal potential, hence, can be used as an effective alternative to the existing synthetic pesticides for the control of Cx. quinquefasciatus.

Simultaneous analysis of artemisinin and flavonoids of several extracts of Artemisia annua L. obtained from a commercial sample and a selected cultivar

Artemisia annua L. (Qinghao) is a promising and potent antimalarial herbal drug. This activity has been ascribed to its component artemisinin, a sesquiterpene lactone that is very effective against drug-resistant Plasmodium species with a low toxicity. Our studies indicate that several flavonoids of A. annua can promote and enhance the reaction of artemisinin with hemin. These data are in good agreement with previous investigations on the in vitro potentiation of antimalarial activity of artemisinin by such flavonoids. As a consequence, in view of a possible use of the phytocomplex rather than pure artemisinin, an HPLC/DAD/MS method is proposed for the simultaneous detection and quantification of both flavonoids and artemisinin. Different extracts, obtained from two different herbal drugs, a commercial sample and a selected cultivar, were analyzed in order to determine which solvents provide the best yields of both artemisinin and flavonoids. Qualitative and quantitative results obtained using an HPLC method are described, which will be useful for developing highly effective herbal drug preparations.

Phytoextract-induced developmental deformities in malaria vector

Larvicidal potential of petroleum ether (Pee), carbon tetrachloride (Cte) and methanol extract (Mee) of Artemisia annua, Chenopodium album and Sonchus oleraceus was observed against malaria vector, Anopheles stephensi Liston. The Pee of A. annua with LC 50 16.85 ppm after 24 h and 11.45 ppm after 48 h of treatment was found most effective, followed by Cte of A. annua and Ch. album, Pee of Ch. album and Mee of A. annua. However, no significant larvicidal activity was observed in Mee of Ch. album and all the three extracts of S. oleraceous. The Pee of A. annua was further investigated for its effect on the metamorphosis and the development of the malaria vector. It influenced the early life cycle of An. stephensi by reducing the percentage of hatching, larval, pupal and adult emergence and also lengthening the larval and pupal periods. The growth index was also reduced significantly. As the extract has remarkable effect on the metamorphosis and high larvicidal potential, it could, therefore, be used as an effective biocontrol agent against the highly nuisant malaria vector.

Lipids from Flowers and Leaves of Artemisia annua and Their Biological Activity

Chemical, chromatographic, and spectral methods were used to show that the main components of the lipid extract of flowers and leaves are free and bound aliphatic and cyclic alcohols (sterols and triterpenols) and essential oil. It was shown that the lipid extract of Artemisia annua has a positive influence on skin metabolism and possesses anti-inflammatory activity.

&lt;i&gt;In vitro&lt;/i&gt; Inhibition of &lt;i&gt;Mycobacterium smegmatis&lt;/i&gt; and &lt;i&gt;Mycobacterium tuberculosis&lt;/i&gt; by Some Nigerian Medicinal Plants

Some Nigerian plants used in traditional medicine to treat tuberculosis and/or some of its symptoms were screened for in vitro activity against Mycobacterium smegmatis and a clinical isolate of Mycobacterium tuberculosis. Only 3 of the 6 crude methanolic extracts of the 6 plant species exhibited inhibitory activities against M. smegmatis, while 5 inhibited the growth of M. tuberculosis. Three and four water extracts inhibited M. smegmatis and M. tuberculosis, respectively. Both methanol and water extracts of Artemisia annua, Pterocarpus erinaceus and Piper guiniense showed inhibitory activities against the two Mycobacteria. Methanol extracts of Anogeissus leiocarpus and Piper guiniense exhibited the highest activity against M. tuberculosis with a minimum inhibitory concentration of 0.1 mg/ml. Key Words: Nigerian medicinal plants, Mycobacterium species, Inhibition East & Central African Journal of Pharmaceutical Sciences Vol.6(1) 2003: 15-19