Ethyl Acetate Root Research Articles

Anticancer Activity of Ethyl Acetate Root Extract of Xanthium strumarium Linn. in N-Nitroso N-Methyl Urea Induced Mammary Carcinoma in Female Sprague-Dawley Rats

The objective of the study was to evaluate the anticancer activity of ethyl acetate root extract of Xanthium strumarium Linn. (EAREXS) in N-Nitroso N-methyl urea (NMU) induced mammary carcinoma in female Sprague-Dawley rats. Thirty virgin female Sprague-Dawley rats of 55 day old were divided into five groups (normal control, disease control, EAREXS 200 mg/kg, EAREXS 400 mg/kg, tamoxifen10 mg/kg) of six animals each. The mammary carcinoma was induced by two intraperitoneal doses of N-Nitroso N-methyl urea (NMU) (50mg/kg/body weight each) in 0.9% NaCl (maintained at 4°C) at 55 day of age and 4 week after the prior administration. The drug EAREXS was administered in two doses, 200 mg/kg, and 400 mg/kg orally and compared with the oral dose of tamoxifen (10 mg/kg). EAREXS 200 mg/kg, 400 mg/kg, and tamoxifen10 mg/kg decreased the tumor incidences by 50%, 66.67%, and 83.33% and the total number of tumors in group by 46.15%, 69.23% and 92.30% respectively, when compared to the disease control. EAREXS 200 mg/kg, 400 mg/kg, and tamoxifen10 mg/kg decreased the average tumor burden by 68.61%, 86.65% and 94.91% and average tumor volume by 66.45%, 82.01%, and 92.98% respectively, when compared to disease control group. The results concluded that EAREXS prevent the mammary carcinogenesis induced by N-Nitroso N-methyl urea in female Sprague-Dawley rats.

Antifungal Activity of the Extractives of Coleus Forskohlii Roots and Forskolin

Hexane, ethyl acetate and methanol extractives of Coleus forskohlii roots exhibit mycelial growth inhibition of Colletotrichum gloeosporioides and spore germination inhibition of Alternaria solani. Forskolin, a diterpene present in C. forskohlii exhibit dose dependent spore germination inhibition of A. solani, this being the first report on the antifungal activity of this compound. Estimation of forskolin in the extractives by HPLC showed that while hexane extractive contained 3.9% forskolin and ethyl acetate extractive contained 4.6% forskolin, while the methanol extractive did not contain forskolin. The results clearly show that forskolin is one of the antifungal compounds present in the plant.

Chemical Composition, Bioactive Components and Antioxidant Activities from Root, Bark and Leaf Lindur

<p>Plants lindur (Bruguiera gymnorrhiza) is one of the plants that have a potential as<br />a source of bioactive compounds for the antioxidant. This study aimed to determine the<br />chemical composition, bioactive compound and determine antioxidant activity of leaves,<br />bark and roots of plants lindur (B. gymnorrhiza). This research was conducted through<br />several stages proximate test, stratified extraction, phytochemical test, and antioxidant<br />activity test with DPPH. The highest yield value resulting by ethanol extract of the leaves<br />is 12,85 % and the lowest was produced by n-hexane root extract that is 0,18 %. The leaves<br />contain high levels of protein, fat, and water which is the higher of the two parts from the<br />other samples. Bark containing ash content (4,12%) and carbohydrates (46,02%). Root has<br />the highest levels of carbohydrates that is 25,91 %. Bioactive components were detected on<br />ethanol and ethyl acetate extracts of the leaves are flavonoids, tannins, phenols, saponins,<br />steroids and tritepenoid. Bark extract ethanol, ethyl acetate and ethanol root bark has<br />bioactive components there are flavonoids, tannins, phenols, saponins, and tritepenoid. </p><p>Ethyl acetate extract of the roots only have bioactive components there are flavonoids,<br />phenols, saponins, and triterpenoids. The potential antioxidant activity produced from<br />ethyl acetate extracts of bark lindur with IC50 values is 14,21 ppm.<br />Keywords: Antioxidants, phytochemicals, roots, ethanol, Bruguiera gymnorrhiza</p>

In vitro Antimicrobial Study for Biological Evaluation of Clerodendrum infortunatum Linn.

To explore a scientific idea, this study was examined for evaluation of antimicrobial potency using root, leaf and stem of ethyl acetate and chloroform extracts of C. infortunatum (Verbenaceae) due to randomly use in traditional medicine to cure common ailments such as intestinal disorder, diarrhea, tuberculosis and respiratory problems etc. The in vitro application was carried out by using disc diffusion, micro broth dilution and serial dilution techniques against clinically important life threatening organisms. All the extracts showed significant inhibitory activity over the bacteria and fungus comparable to the standard drug tetracycline and fluconazole. The maximum average diameter zone of inhibition was recorded to bacterial strains against B. megaterium, S. typhi, K. pneumoniae and to fungi against A. niger and C. albicans. The MIC values of ethyl acetate and chloroform root extract were determined 64 µg/ml to B. subtilis, and K. pneumoniae; to S.-β-haemolyticus and S. typhi for ethyl acetate extracts, 128 µg/ml to S. aureus, and E. coli for both ethyl acetate and chloroform root extracts but only S. typhi and S.-β-haemolyticus for chloroform extract. The findings evidently appear promising antibacterial and antifungal properties of C. infortunatum against antagonistic pathogens. Leaf possess quite potent activity than root and stem specially root extract > leaf extract > stem extract. One of the more significant achievements of this study to follows and covers the most recent and important patents WO2009075290 (2009) which deals on yeast having immunopotentiating effect and food or feed. This study serves as basis for further research to lead compounds to be isolated so that may be as a template for the implications of these results for bioactivity and drug discovery potential of herbal products are discussed.

PHYTOCHEMICAL AND BIOLOGICAL STUDY OF (Senecio glaucus subsp. coronopifolius ) (MAIRE) C. ALEXANDER GROWING IN EGYPT

Senecio glaucus subsp. coronopifolius (Maire) C. Alexanderis wild annual herb distributed in the Egyptian deserts. Total phenolic and flavonoid content of plant root were determined using both HPLC and colorimetric analysis. Syringic acid and hesperidin (1378.802 and 6638.247 mg / 100 gm. dried plant root powder, respectively) were of the highest concentration compounds resulted from HPLC analysis of total phenolic and flavonoid content. The colorimetric estimation of total phenolic and flavonoid content resulted in concentration of (98.23 ± 0.28 mg/gm. expressed as Gallic acid equivalent (GAE) and35.9± 0.17 mg/gm.expressed as quercetin equivalent (QE), respectively). GC-MS analysis of un-saponifiable matters and fatty acid methyl esters of the plant leaves indicated that octacosane (11.85%) and linolenic acid methyl ester (31.07%) (poly- unsaturated fatty acid) were the major identified compounds, respectively. The DNA of the plant was analyzed using twelve random decamer primers. A total of 52 random amplified polymorphic DNA (RAPD) markers were identified. Root extracts (ethyl acetate, acetone and methyl alcohol) were subjected to determine the antimicrobial behavior and also their cytotoxic activity, by using (3- (4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (MTT) assay against colon carcinoma cell lines (HCT-116). Among the fore mentioned extracts, root ethyl acetate extract gave appreciable antibacterial and antifungal behavior and also had promising cytotoxic activity with IC50 = 7.39 ±1.2 µg/ml. Root methyl alcohol extract showed antioxidant activity with IC50 = 79.57 ± 0.74 µg/ml, using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay.

KOMPOSISI KIMIA DAN AKTIVITAS ANTIOKSIDAN AKAR, KULIT BATANG DAN DAUN LINDUR

Plants lindur (Bruguiera gymnorrhiza) is one of the plants that have a potential as a source of bioactive compounds for the antioxidant. This study aimed to determine the chemical composition, bioactive compound and determine antioxidant activity of leaves, bark and roots of plants lindur (B. gymnorrhiza). This research was conducted through several stages proximate test, stratified extraction, phytochemical test, and antioxidant activity test with DPPH. The highest yield value resulting by ethanol extract of the leaves is 12,85 % and the lowest was produced by n-hexane root extract that is 0,18 %. The leaves contain high levels of protein, fat, and water which is the higher of the two parts from the other samples. Bark containing ash content (4,12%) and carbohydrates (46,02%). Root has the highest levels of carbohydrates that is 25,91 %. Bioactive components were detected on ethanol and ethyl acetate extracts of the leaves are flavonoids, tannins, phenols, saponins, steroids and tritepenoid. Bark extract ethanol, ethyl acetate and ethanol root bark has bioactive components there are flavonoids, tannins, phenols, saponins, and tritepenoid. Ethyl acetate extract of the roots only have bioactive components there are flavonoids, phenols, saponins, and triterpenoids. The potential antioxidant activity produced from ethyl acetate extracts of bark lindur with IC50 values is 14,21 ppm. Keywords: Antioxidants, phytochemicals, roots, ethanol, Bruguiera gymnorrhiza

Activity of Fabaceae species extracts against fungi and Leishmania: vatacarpan as a novel potent anti-Candida agent

Leishmaniasis and fungal infection treatment efficacy is limited by toxicity and ever increasing resistance to available drugs, requiring development of alternative compounds. The richness of Cerrado plant antimicrobial secondary metabolites justifies screening of Fabaceae species extracts: Enterolobium ellipticum Benth., Sclerolobium aureum (Tul.) Baill. and Vatairea macrocarpa (Benth.) Ducke, against Leishmania (Leishmania) amazonensis, yeasts and dermatophytes. Among the 26 extracts tested, more than 50% of the total demonstrated significant antifungal activity in comparison to the drug controls (minimal inhibitory concentration 0.12 to ≤31.25μg/ml). Six extracts capable of complete parasitic growth inhibition had the inhibitory concentration index for 50% values from 9.23 to 78.65μg/ml. The results led to the selection of the V. macrocarpa ethyl acetate root bark extract for chemical fractionation. This plant, traditionally referred to as angelim-do-cerrado or maleiteira, is used to treat superficial mycoses in Amazonia. A previously unreported pterocarpan vatacarpan together with the known compound musizin was isolated. Vatacarpan demonstrated a minimal inhibitory concentration value of 0.98μg/ml against Candida albicans ATCC 10231, and thus comparable or superior to fluconazole and amphotericin B. The results add to literature's information the ability of pterocarpans to act as antimicrobial agents.

Evaluation of antioxidant and anticancer activity of extract and fractions of Nardostachys jatamansi DC in breast carcinoma.

BackgroundNardostachys jatamansi DC is a Himalayan medicinal herb that has been described in various traditional systems of medicine for its use in cancer. In view of its traditional claims, and chemical constituents, antioxidant and anticancer activities were evaluated in breast carcinoma.MethodsPetroleum ether (NJPE), methanol extract (NJM) and subsequent diethyl ether (NJDE), ethyl acetate (NJEA) and aqueous (NJAQ) fractions of roots and rhizomes of N. jatamansi were prepared. Total phenolic, flavonoid content, and antioxidant activities were determined using suitable methods. Antiproliferative activity was assessed in estrogen receptor (ER)-positive (MCF-7) and ER-negative breast carcinoma (MDA-MB-231) cells by MTT and SRB assay. Cell cycle analysis, Hoechst staining, and clonogenic assay were employed to determine the mode of antiproliferative and pro-apoptotic activity in MDA-MB-231 cells.ResultsNJM/fractions exhibited prominent antioxidant activity with significant correlation between phenolic content and ABTS (IC50) scavenging (R = −0.9680, P < 0.05), and total antioxidant capacity (R = 0.8396, P > 0.05). In MTT assay, NJM exhibited the highest antiproliferative activity (IC50: 58.01 ± 6.13 and 23.83 ± 0.69 μg/mL in MCF-7 and MDA-MB-231 respectively). Among the fractions, NJPE and NJDE were found to be most potent in MCF-7 (IC50: 60.59 ± 4.78 μg/mL) and MDA-MB-231 (IC50: 25.04 ± 0.90 μg/mL) cells respectively. Statistical analyses revealed NJM and NJDE exhibited significantly higher (P < 0.05) cytotoxicity in MDA-MB-231 cells. Cell cycle analysis demonstrated that NJM, NJPE and NJEA caused G2/M arrest while NJDE caused G0/G1 phase arrest in MDA-MB-231 cells. Further, NJM/fractions induced significant (P < 0.001) cell death by apoptosis characterized by apoptotic morphological changes in Hoechst staining and inhibited long-term proliferation (P < 0.001) of MDA-MB-231 cells in clonogenic assay. Lupeol and β-sitosterol were identified as anticancer principles in NJM/fractions by HPTLC.ConclusionOur results suggest that NJM/fractions possess significant antiproliferative potential which is mediated through cell cycle perturbation and pro-apoptotic effects in MDA-MB-231 cells. Moreover, this study highlights the antioxidant potential of NJM/fractions which can be attributed to the presence of phenols. NJDE emerged as the most potent fraction and further mechanistic and phytochemical investigations are under way to identify the active principles.

FREE RADICAL-SCAVENGING POTENTIAL OF DIFFERENT EXTRACTS FROM AZIMA TETRACANTHA LAM.

Oxidative stress resulted from free radicals and reactive oxygen species are associated with man y diseases. Worldwide several studies are directed towards finding natural antioxidants of plant origin. Plants containing phenolic compounds have been reported to pos sess strong antioxidant activities . The aim of the present study was to investigate the p hytochemical constituent of hexane, chloroform, ethyl acetate and methanolic root extracts and in vitro antioxidant activities of A zima tetracantha Lam. The different extract s of A. tetracantha root were studied for antioxidant potential by using different in vitro assays such as inhibition of DPPH, ABTS + , Hydroxyl radical and superoxide anions. The total phenolic contents and ferric reducing antioxidant power of the extracts were also determined by using standard phytochemical reaction methods. Butylated h ydroxyl toluene (BHT), Vitam in C and ascorbic acid were taken as standards. The different extract s of A. tetracantha root showed good dose dependant free radical scavenging activity in all the models. The total phenolic content of hexane, chloroform, ethyl acetate and methanolic root extracts was found to be 3 .30±0.20, 4.20±0.20, 6.30±0.20 and 8.80±0.20 mg GAE /g respectively . Ferric reducing antioxidant power was also found to be increase d with increase in extracts concentrations. All the results of the in vitro antioxidant assays revealed the free radical scavenging potential of A. tetracantha root and compared with standard antioxi dants. These antioxidant activities may be endorsed to its high free radical scavenging and phenolic contents. Thus, our findin gs provide evidence that A. tetracantha is a potential source of natural antioxidants.

Larvicidal Potentiality of Root Extracts of &lt;i&gt;Annona reticulata&lt;/i&gt; Linn. Against the Filarial Vector &lt;i&gt;Culex quinquefasciatus&lt;/i&gt; Say (Diptera: Culicidae)

Present study was undertaken to evaluate the mosquito larvicidal potentiality of root extracts of Annona reticulata Linn. ( A. reticulata ), family Annonaceae against filarial vector Culex quinquefasciatus Say, 1823 ( Cx. quinquefasciatus ). Petroleum ether, hexane, and ethyl acetate root extracts of the plant were examined to established their larvicidal potentiality against 3 rd instar larvae of Cx. quinquefasciatus mosquito species. Crude and only ethyl acetate root extracts of the plant at different concentrations were examined on 1 st - 4 th instars larvae of Cx. quinquefasciatus . LC 50 value of ethyl acetate extract after 24 h of exposure against 3 rd instar larvae of Cx. quinquefasciatus were also examined on some non target organisms. Phytochemical analyses of the root extracts of A, reticulata were also investigated. 100% mortality were noticed at very low concentrations of petroleum ether, hexane, and ethyl acetate root extracts of A. reticulata against 3 rd instar larvae of Cx. quinquefasciatus . LC 50 and LC 90 values of ethyl acetate root extract was lower than petroleum ether and hexane root extracts after 24, 48, and 72 h of exposure. Crude and ethyl acetate root extracts showed excellent larvicidal activity against Cx. quinquefasciatus mosquito species. 100% mortality was observed at very low concentration for crude and ethyl acetate root extracts. LC 50 and LC 50 values gradually decreased with time of exposure for crude and ethyl acetate root extracts of A. reticulata . No mortality and sluggishness were observed in control experiments as well as on examined non target organisms. Phytochemical analyses of root extracts of the plant revealed the presence of different secondary metabolites. So crude and ethyl acetate root extracts of A. reticulata can effectively be used for larval mortality against Cx. quinquefasciatus mosquito species at a very low concentration.

Evaluation of anti-oxidant and acetylcholinesterase activity and identification of polyphenolics of the invasive weed Dittrichia viscosa.

The bioactive metabolites derived from weeds have attracted the interest of the food and pharmaceutical industries due to their health benefits. To evaluate the anti-oxidant and acetylcholinesterase activity of Dittrichia viscosa extracts and characterise the polyphenolic metabolites using the LC coupled with diode-array detection (DAD) and positive mode electrospray ionisation (ESI) MS method with a view to evaluating the exploitation potential of this invasive weed. Roots and aerial parts of D. viscosa were extracted with solvents of increasing polarity and their major polyphenolic metabolites were identified by LC - DAD/ESI(+)/MS. The total phenolic content of the extracts was determined using the Folin-Ciocalteu method, while their anti-oxidant activity was evaluated on the basis of their ability to scavenge the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydrogen peroxide. Thin-layer chromatography was used to screen for acetylcholinesterase inhibitors. Stem extracts gave the highest phenolic content, whereas the roots showed the lowest content. Twenty-five polyphenolic constituents of the extracts were tentatively characterised according to their MS and UV spectroscopic data. Among the extracts studied, roots-ethyl acetate and flowers-diethyl ether revealed the highest activity according to the DPPH and chemiluminescence assays respectively. The metabolic profile of D. viscosa was studied and the structures of the major polyphenolic metabolites were tentatively assigned based on their MS and UV-vis spectra. The extracts exhibited high levels of anti-oxidant and acetylcholinesterase inhibitory activity and the inhibitors are probably localised mainly in flowers and roots.

In-Vitro and in-Silico characterization of Sophora interrupta plant extract as an anticancer activity.

Sophora interrupta belongs to the family of Fabaceae and the species in this genus have a diverse medicinal importance as a folk medicine for preventing many ailments including cancer. In order to evaluate the anticancer activity of S.interrupta, we have performed in vitro anti-oxidant, anti-inflammatory, anti-proliferative, and cell based anticancer activity in MCF-7 and PC-3 cell lines. Secondary metabolites of S.interrupta were used to identify anticancer compounds using Open Eye software. The antioxidant activity of the S.interrupta root ethylacetate (SEA) extract at 100 µg/ml is equal to that of ascorbic acid at 50 µg/ml. The antiinflammatory activity of SEA is half of that of diclofenac at 50 µg/ml. Anticancer activity was detected by measuring the mitochondrial dehydrogenase activity (MTT assay). The half maximal inhibitory concentrations (IC50) for MCF-7 and PC-3 cell lines are 250 and 700 µg/ml respectively. This was supported by the morphological changes such as membrane blebbing, cell detachment and rounded cell morphology when compared to the parental cells. In addition, we observed few green cells (live) over red cells (dead) based on the uptake of acridine orange and ethidium bromide dyes. Kaempferol-3-O-b-D-glucopyranoside, a Secondary metabolite of S.interrupta form 6 hydrogen bond interactions with Arg 202, Gln 207, Gly 227, Gly 229, Thr 231 and Ala 232 human DEAD box RNA helicase, DDX3 protein and is equivalent to crystal structure of adenosine mono phosphate to DDX3. Overall, it suggests that the SEA extract has anticancer compounds, and it can be used to enhance death receptor mediated cancer cell death.

Antimicrobial activity of Capparis spinosa as its usages in traditional medicine

Summary Capparis spinosa roots and fruits are traditionally used for the treatment of diseases such as diarrhea and hemorrhoids. In the 10th century, farmers also used aqueous extract from its roots for disinfecting their plants’ seeds. Due to widespread use of this plant in traditional medicine, we evaluated different extracts (aqueous, methanol, ethanol and ethyl acetate) of fruits and roots of this plant against bacteria and fungi causing infections in plants or humans. The total phenolic and flavonoid content of extracts from fruits and roots and the antioxidant activity were evaluated. The antimicrobial activity was evaluated by microdilution assay in fluid medium. Among different extracts from roots and fruits, root ethyl acetate and fruit methanol extracts had higher total phenolic content, while fruit ethyl acetate extract contained higher total flavonoid content. The higher antioxidant activity was shown for roots ethanol extract (IC50=88 μg/ml). As its traditional uses, roots aqueous extract from C. spinosa exhibited inhibitory effect against bacteria and fungi with the exception of Pseudomonas aeruginosa. With the exception of fruit aqueous extract, other extracts (methanol, ethanol and ethyl acetate) from fruit and root exhibited good activity against microorganisms, especially fungi. This study reveals the traditional uses of roots and fruit extracts as antimicrobials agent.

Centrifugal partition chromatography for antibacterial bio-guided fractionation of Clidemia hirta roots

Clidemia hirta is an invasive shrub used in traditional medicine to treat some bacterial infections. In order to identify the molecules involved in this antimicrobial activity and to consider C. hirta as a potential source of preservative molecule usable in cosmetic applications, an antibacterial bio-guided screening of C. hirta root ethyl acetate extract was developed. Centrifugal partition chromatography performed using an Arizona N (heptane/ethyl acetate/methanol/water 1:1:1:1 v/v/v/v) system in descending mode, allowed to recover three fractions. For three samples of C. hirta root, independently collected, each crude extract and each fraction were tested against strains recommended in the current regulatory method for cosmetic preservative to evaluate the antimicrobial activity and on keratinocyte and fibroblast cells to verify the absence of skin cell toxicity. The first fraction showed an interesting antibacterial activity but also sometimes skin cell toxicity. Hydrolysable tannins, derivatives of ellagic acid were identified using mass spectrometry in this fraction. The second fraction was both antibacterial and not cytotoxic. Mass spectrometry and NMR experiments confirmed the isolation of only one compound in this fraction which was identified as arjunolic acid.

Phagodeterrence, larvicidal and oviposition deterrence activity of Tragia involucrata L. (Euphorbiaceae) root extractives against vector of lymphatic filariasis Culex quinquefasciatus (Diptera: Culicidae)

ObjectiveTo evaluate phagodeterrence or biting deterrence, larvicidal and oviposition deterrence activity of Tragia involucrata (T. involucrata) root extractives against Culex quinquefasciatus (Cx. quinquefasciatus). MethodsPhagodeterrence test was done using five different solvents namely petroleum ether, benzene:ethyl acetate (1:1 v/v), chloroform:methanol (1:1 v/v), acetone and absolute alcohol against the females of Cx. quinquefasciatus. Effective deterrence was recorded. The larvicidal bioassay was performed by solvent extractives of T. involucrata roots against 1st, 2nd, 3rd and 4th instars larval forms of Cx. quinquefasciatus. Mortality rates were recorded after 24, 48 and 72 h of exposure followed by justification of LC50 and LC90 values at different concentrations. Finally, the larvicidal assay was ended up by recording the health belongings of non-target water fauna. Oviposition deterrence activity of root extractives was examined against the gravid females of Cx. quinquefasciatus. ResultsAbout 92.85% phagodeterrence was noticed at the level of 4% of chloroform:methanol (1:1 v/v) for the very 1st hour of exposure. Entire larval populations of 1st and 2nd instars were subjected to diminution following the treatment of 0.4% and 0.5% of benzene:ethyl acetate (1:1 v/v) root extractives respectively. While 92.33% and 78.33% mortality were recorded for 3rd and 4th instars larval forms respectively. Mean effective oviposition deterrence was found to be 97.85% at 2.5% concentration level of chloroform:methanol (1:1 v/v) root extractives. ConclusionsThis study is a pioneer attempt to establish phagodeterrence, larvicidal and oviposition deterrence activity of T. involucrata root extractive.

Evaluation of in Vitro Antioxidative, Cytotoxic and Apoptotic Activities of Rheum ribes Ethyl Acetate Extracts

Rheum species are medicinally important plants due to the presence of anthracene derivatives. This study was designed to determine the antioxidative, cytotoxic and apoptotic properties of Rheum ribes shoot and root ethyl acetate extracts using human promyelocytic leukemia (HL-60) cell line as a model system. R. ribes shoot and root dry powder samples were prepared and extracted with ethyl acetate. The extracts were revealed to be a potential scavenger of DPPH radicals (IC50 value of 206.28 μg/ml for shoot and 10.92 μg/ml for root) and the chemical composition of the extracts was quantified by colorimetric determination of total phenol (GAE) and flavonoid (CAE) contents. HL–60 cells were cultured in the presence of various concentrations of extracts up to 72 h. R. ribes inhibited the survival of HL-60 cells in a concentration- and time-dependent manner, shown by XTT assay. R. ribes caused HL-60 cells apoptosis via formation of phosphatidylserine externalization, as evidenced by flow cytometry. Exposure of HL-60 cells to higher concentrations of extracts for 72 h resulted in a shift of 87% of the cell population from normal to the early/late apoptotic stage. These findings suggest that Rheum ribes ethyl acetate root extracts exhibits potential antioxidant and cytotoxic properties against HL-60 cells better than shoot extracts and exert their toxicity via induction of apoptosis.

Antibacterial activity of the roots, stems and leaves of Alchornea floribunda

Ethnopharmacological relevanceAlchornea floribunda Müll. Arg. is used in traditional medicine across Africa for the treatment of bacterial, fungal, parasitic and inflammatory disorders. Aim of the studyTo evaluate the antibacterial activity of the crude extracts of different plant parts in order to provide a scientific rationale for the proposed broad efficacy of Alchornea floribunda in the treatment of bacterial infections. Materials and methodsExtracts of roots, stems and leaves were prepared using solvents of various polarities in order to extract a wide range of phytochemicals. The antibacterial activity of these crude extracts was evaluated by micro-dilution assay, against Gram-positive (i.e. Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus saprophyticus) as well as Gram-negative (i.e. Escherichia coli, Klebsiella pneumoniae, Moraxella catarrhalis and Proteus mirabilis) bacteria. ResultsGenerally, the ethanol (EtOH), methanol (MeOH), ethyl acetate (EtOAc) and chloroform (CHCl3) extracts demonstrated the best activities, with the leaves exhibiting the highest average activity for six of the eight pathogens. Of these, the ethanolic leaf extract was the most active against Staphylococcus aureus with an MIC value of 50µg/mL. Some other notable activity was observed for the ethyl acetate and chloroform root extracts against Staphylococcus aureus (50µg/mL), and for selected stem extracts against Staphylococcus aureus (50µg/mL), Klebsiella pneumoniae (63µg/mL) and Staphylococcus saprophyticus (63µg/mL). ConclusionThis study demonstrates the promising antibacterial activity of Alchornea floribunda against both Gram-positive and Gram-negative bacteria responsible for gastrointestinal, skin, respiratory and urinary ailments, and validates its use in the ethnopharmacology of the region.

In vitro analysis on bactericidal screening and antioxidant potentiality of leaf and root extracts of Thottea siliquosa (Lam.) Ding Hou. An ethnobotanical plant

ObjectiveNatural products of plant origin are potential source of novel antimicrobial and antioxidative agents. Thottea siliquosa (Lam.) Ding Hou. (T. siliquosa). A medicinal herb used by local tribals for treating various ailments. The present study aims at the phytochemical screening, GC-MS analysis, In vitro antibacterial activity and antioxidant potentiality of root and leaf extracts of T. siliquosa. MethodsHot continuous Soxhlet extraction, GC-MS analysis, antibacterial analysis by disc diffusion, microdilution assay and antioxidant potentialities by hydroxyl radical and nitric oxide radical scavenging. The data was statistically analyzed. ResultsPhytochemical screening of the ethyl acetate and methanolic extract of leaf and root revealed the presence of phenols, alkaloids, tannins and saponin. The extract revealed a pool of phytochemicals by comparison with authentic standards from spectral library. Both the extracts has shown their broad spectrum of inhibition against the selected bacteria Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumonia compared with standard antibiotic drug streptomycin. The extracts showed antioxidant activity by scavenging of free radicals such as hydroxyl and nitric oxide. The IC50 values of the ethyl acetate extracts leaf and root and standard in this assay were 167.5±0.67, 99.4±1.2, 192±2.5 μg/mL respectively. Similarly those methanolic extracts of leaf and root were 269.5±0.89 and 289.1±2.66 μg/mL respectively. Similarly, ethyl acetate and methanolic extracts also caused a moderate dose-dependent inhibition of nitric oxide with an IC50 range 65.5±1.55 to 148 ±3.09 μg/mL. The inhibitory activities were found to be dose dependent. ConclusionThe present study provides evidence that ethyl acetate and methanol extract of leaf and root of T. siliquosa are potential source of natural antioxidants and bactericidal nature. It is essential that research should continue to isolate and purify the bio active components of this natural plant and use in drug discovery and development.

Tyrosinase modulation by five Rwandese herbal medicines traditionally used for skin treatment

Ethnopharmacological relevanceTraditional herbal medicines provide an interesting, largely unexplored source for the development of potential new drugs and skin-care cosmetics. Some herbal extracts are known to be inhibitors of melanin formation, sometimes more potent than the classical inhibitors, hydroquinone/arbutin or kojic acid, and are not associated with melanocytes cytotoxicity or mutagenicity. Such plants are used in traditional medicine in many countries, particularly in Africa, for skin lightening. Aim of the studyTo evaluate in vitro the ability of Rwandese medicinal plants, traditionally used for the treatment of skin (discoloration and attenuation of discolored spots), to modulate pigmentation and tyrosinase activity. Materials and methodsBased on an ethnopharmacological survey, five herbs [Brillantaisia cicatricosa Lindau (Acanthaceae), Chenopodium ugandae (Aellen) Aellen (Chenopodiaceae), Dolichopentas longiflora Oliv. (Rubiaceae), Protea madiensis Oliv. (Proteaceae) and Sesamum angolense Welw. (Pedaliaceae)] were selected. Twenty-seven extracts, obtained by treating the herbs with increasing polarities solvents, were investigated for their effects on cell viability (MTT test) and on pigmentation: inhibition of the enzyme tyrosinase (colorimetry of reaction products, measurement of enzyme activity, TLC-autography; studies on crude cellular extracts obtained from normal melanocytes and on a mushroom tyrosinase) and measurement of melanogenesis by human melanoma cells. ResultsNone of the tested plant extracts were cytotoxic on tested human melanoma cell lines, except for Dolichopentas longiflora (IC50 of leaves n-hexane extract, 4μg/ml for MM028 and 4.5μg/ml for MM001; IC50 of roots ethyl acetate extract, 0.8μg/ml for MM028 and 3.9μg/ml for MM001). Almost all extracts inhibited melanogenesis in a melanoma whole cells overall pigmentation assay, a model reflecting the entire cycle of melanogenesis. All the Protea madiensis extracts quite strongly inhibited melanogenesis and, surprisingly, one of the Dolichopentas longiflora leaves extracts was found to increase melanogenesis. These results were confirmed by the modulation of pigmentation reactions by crude cellular extracts obtained from normal melanocytes; interestingly, one of the extracts (Dolichopentas longiflora ethyl acetate extract) is even more active (61% at 500μg/ml) than kojic acid (<3% at 142μg/ml and 68% at 1421μg/ml). In a mushroom tyrosinase inhibition assay, data obtained on some extracts fairly agree with pigmentation inhibition measured on melanocytes proteins as, for example, the methanol extract of Protea madiensis. While a few others extract display discording data, this probably reflects either differences between human and mushroom tyrosinase, interference with melanocytes enzymes at later steps than tyrosinase or the simultaneous presence of compounds with conflicting activities in a given extract. ConclusionsEthnopharmacological data represent an efficient approach to discover active herbs. Some of the selected medicinal plants clearly show potent tyrosinase inhibitions while one extract significantly increases cell pigmentation; one extract contains potent growth melanocytes inhibitors.

Antimalarial and safety evaluation of extracts from Toddalia asiatica (L) Lam. (Rutaceae)

Ethnopharmacological relevanceToddalia asiatica (L) Lam. (Rutaceae) is a medicinal plant traditionally used in Kenya by many communities for the treatment of malaria and other ailments. All parts of the plant are claimed to have medicinal value, but the root bark in particular is believed to be more potent. Decoctions or infusions of the roots are taken orally to treat malaria, fever and stomach ache. Aim of the studyTo evaluate antimalarial activity of aqueous and organic extracts prepared from Toddalia asiatica and determine in vitro and in vivo safety of the extracts. Materials and methodsAqueous, ethyl acetate, hexane and methanol extracts were obtained from Toddalia asiatica root bark, fruits and leaves. In vitro antiplasmodial activity was done using chloroquine-sensitive (D6) and chloroquine-resistant (W2) Plasmodium falciparum strains and the concentration causing 50% inhibition of radioisotope incorporation (IC50) was determined. In vivo assay was done by administering mice infected with Plasmodium berghei four consecutive daily doses of the extracts through oral route following Peters 4-Day suppressive test. The percentage suppression of parasitaemia was calculated for each dose level by comparing the parasitaemia in untreated control with those of treated mice. Quinine hydrochloride was used as positive control while double distilled water or 20% Tween-80 was used as a negative control. In vivo acute toxicity was determined in mice using standard procedures. In vitro cytotoxicity assay was carried out using actively dividing sub-confluent Vero cells. ResultsInhibitory concentrations of ethyl acetate extract of Toddalia asiatica fruits showed high activity against chloroquine resistant (W2) strains of Plasmodium falciparum (IC50=1.87μg/ml), followed by root bark aqueous extract (IC50=2.43μg/ml). Tested in vivo against Plasmodium berghei, the fruit ethyl acetate extract (500mg/kg) and root bark aqueous extract (250mg/kg) reduced malaria parasitaemia by 81.34% and 56.8% respectively. Higher doses were found to be less effective in vivo. Acute toxicity and cytotoxictiy of the tested extracts, with the exception of hexane extract from the roots, showed LD50>1000mg/kg and CC50>100μg/ml respectively. ConclusionsThe results obtained contribute to the validation of traditional use of Toddalia asiatica and provides in vivo and safety data of the plant extracts tested for the first time. Ethyl acetate extract of the fruits was active against chloroquine resistant Plasmodium falciparum as well as against Plasmodium berghei. These findings confirm the suitability of Toddalia asiatica as a good candidate for further tests to obtain a prototype for antimalarial medicine.