Ethyl Acetate Fraction Of Extract Research Articles

Piper betle leaves extract adversely affect motility of Pseudomonas aeruginosa

Pseudomonas aeruginosa is an opportunistic pathogen that orchestrate its disease manifestation via a complex communication system known as quorum sensing (QS). Under the regulation of this complex network of QS system, is a myriad of virulence factors including the flagella and pili that contribute to its motility. To establish an infection in a host, P. aeruginosa must first attach itself to host cells with the assistance of flagella and pili and it would be a mere impossible for P. aeruginosa to initiate and establish an infection in the host without those features. In this study, we investigated the motility of P. aeruginosa under the influence of Piper betle leaves extract by growing the bacteria on a semi solid medium and compared the growth spread of the pathogen against the untreated sample. There are three types of motilities that were being investigated, which were swimming, swarming and twitching. In all three assays, a fraction of ethyl acetate extract of P. betle leaves were found to reduce the motility of P. aeruginosa with greatest defect in swimming at more than 80% reduction as compared to the untreated sample. Meanwhile, the fraction partially decreased the swarming and twitching of P. aeruginosa by 58% and 50% reduction respectively, in comparison to the untreated sample. The exhibited defect in the motility of P. aeruginosa proves the anti-QS property of the fraction of ethyl acetate extract of P. betle leaves and could be developed as a potential therapeutic agent against the pathogen.

Anti-type I allergic effects of Jing-Fang powder extracts via PI3K/Akt pathway in vitro and in vivo

Jing-Fang powder (Schizonepeta tenuifolia Briq. and Saposhnikovia divaricata (Turcz.) Schischk.) was used to treat chronic bronchitis, asthma and chronic urticaria. Based on the preliminary results of screening research on the antiallergic effective parts of Jing-Fang powder, its ethyl acetate extract fractions (JFEE) and isolate D (JFEE-D) showed the best anti-allergic effect. RBL-2H3 cell activation degranulation model and mice passive cutaneous anaphylaxis (PCA) reaction model were used to investigate the effects and mechanisms of JFEE and JFEE-D on IgE-mediated type I allergic reactions. LC–MS was utilized to determine the composition of JFEE and JFEE-D. We found that JFEE and JFEE-D significantly reduced β-HEX, histamine, IL-4, IL-6 levels in cell supernatants, and improved the degree and morphology of cell degranulation. JFEE and JFEE-D significantly inhibited the increase of ear vascular permeability and abnormal increase of serum IgE, TNF-α, IL-6 levels. JFEE and JFEE-D inhibited mRNA expression of PI3K and Akt and down-regulated protein expression of PI3K, Akt, p-Akt, and PLCγ1 in sensitized RBL-2H3 cells. The combined use of JFEE and JFEE-D with pathway inhibitor Wortmannin revealed synergistic down-regulation of PI3K, Akt, and p-Akt protein expression. The combined use of pathway agonist IGF-1, JFEE and JFEE-D down-regulated increase of p-Akt/Akt protein expression. Moreover, JFEE and JFEE-D significantly inhibited protein expression of PI3K, p-Akt and PLCγ1 in PCA model mice. These results show that JFEE and JFEE-D inhibit type I allergic reactions by inhibiting PI3K/Akt signaling pathway.

A secondary metabolite with in vitro radical scavenging activity from endolichenic fungus Daldinia eschscholzii found in lichen, Parmotrema sp. in Sri Lanka

Endolichenic fungi, an unexplored group of microorganisms, are a promising source of bioactive compounds. Secondary metabolites were isolated from the chloroform fraction of crude ethyl acetate extract of endolichenic fungus Daldinia eschscholzii inhabiting the lichen, Parmotrema sp. in Sri Lanka. Two pure compounds, 1 and 2 were isolated and the structures were identified using 1H-, 13C-, 2D- nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS) data. Compound 1 did not show any radical scavenging activity in DPPH assay. Compound 2, identified as 8-methoxynaphthalen-1-ol showed strong radical scavenging ability in the DPPH assay with a half maximal inhibitory concentration (IC50) of 10.2 ± 5.8 µg/ mL. The activity of compound 2 was higher than that of the standard, butylated hydroxy toluene (BHT).

Antimalarial Activity of Ethyl Acetate Extract and Fraction of Bidens pilosa against Plasmodium berghei (ANKA).

Background Malaria is one of the most critical diseases causing about 219 million cases worldwide in developing countries. The spread and development of resistance against chemical antimalarial drugs is one of the major problems associated with malaria control. The present study was to investigate the antimalarial efficacy of ethyl acetate extract and one fraction of Bidens pilosa in vivo in order to support the usage of this plant by traditional healers to treat malaria. Methods The extracts were prepared by maceration of B. pilosa leaf powder in ethyl acetate. The liquid filtrate of the extract and the best in vitro antiplasmodial fraction using HPLC were concentrated and evaporated using a rotavapor under vacuum to dryness. The antimalarial activity of B. pilosa plant products were evaluated in vivo against Plasmodium berghei infected mice according to the Peter and Rane test. The antimalarial efficacy of the a selected crude extract (ethyl acetate extract) was evaluated at 125, 250, and 500 mg/kg, while a selected fraction from ethyl acetate extract (fraction 12) was evaluated at 62.5 and 125 mg/kg. Blood from experimental animals was collected to assess hematological parameters. Results The crude extract of ethyl acetate and fraction 12 demonstrated 100% in vivo parasite suppressive activity at doses of 500 mg/kg and 125 mg/kg, respectively, for the crude extract and fraction 12. The mice treated with 250 and 500 mg/kg had their parasitemia (intraerythrocytic phase of P. Berghei) drop considerably, disappearing by the 8th day in mice receiving 500 mg/kg. The ethyl acetate extract of B. pilosa, fraction 12 showed an even higher antiplasmodial activity. By the 5th day of the experiment, the treatment led to a modification of hematological parameters in mice. The chloroquine (5 mg/kg), fraction 12 (125 mg/kg), and the crude extract (500 mg/kg) groups all survived the 30 days of the experiment, while the negative control group registered 100% of the deaths. Conclusion This study scientifically supports the use of Bidens pilosa leaves in the traditional treatment of malaria. However, the mode of action and in vivo toxicity of the plant still need to be assessed.

AKTIVITAS ANTIBAKTERI FRAKSI DAUN CABE RAWIT (Capsicum frutescens L.) TERHADAP BAKTERI Staphylococcus aureus

ABSTRACTStaphylococcus aureus is a normal bacterial flora found on the skin and mucous membranes of humans. Staphylococcus aureus bacteria can also overgrow and invade under certain conditions, act as pathogenic bacteria that cause various diseases or disorders in the body, one of which is a respiratory tract infection. The cayenne pepper plant is widely used by people in the cayenne pepper fruit section as a spice ingredient in various traditional dishes, however, the cayenne pepper leaves are still not widely used. Cayenne pepper leaves contain saponin and phenol group compounds, namely flavonoids, which have inhibitory activity against Staphylococcus aureus bacteria. The purpose of this study was to determine the most active fraction inhibiting the growth of staphylococcus aureus bacteria in vitro. It carried testing the antibacterial activity of the test plants out by the solid diffusion method. The results showed that the ethyl acetate extract fraction from cayenne pepper leaves (Capsicum frutescent L.) Was the most active antibacterial fraction against staphylococcus aureus bacteria with a minimum inhibitory level (MIC) of 10% w / v with a very strong category.

In-vitro Bactericidal Activity of a Novel Plant Source Plumeria pudica against Some Human and Fish Pathogenic Bacteria.

The only remedy for up surging problem of antibiotic resistance is the discovery of antibacterial agents of natural origin. The present study was aimed at finding antibacterial potential of crude and solvent extracts of mature leaves of Plumeria pudica. Antibacterial activity of three different solvent extracts were evaluated in four human and four fish pathogenic bacteria by measuring the zone of inhibition and determining Minimum Inhibitory Concentration and Minimum Bactericidal Concentration values. Standard antibiotics were used as positive control. Preliminary phytochemical screening of most effective extract i.e., ethyl acetate extract, Fourier Transform Infra Red analysis and GC-MS analysis of the Thin Layer Chromatographic (TLC) fraction of ethyl acetate extract were done meticulously. All experiments were done thrice and analyzed statistically. Crude leaf extracts and solvent extracts caused good inhibition of bacterial growth in all selected bacteria. Ethyl acetate extract showed highest inhibition zones in all tested strains with maximum inhibition (19.50±0.29 mm) in Escherichia coli (MTCC 739). MBC/MIC of the extracts indicated that all three solvent extracts were bactericidal. Preliminary phytochemical tests revealed the presence of tannins, steroids and alkaloids and FT-IR analysis revealed presence of many functional groups namely alcoholic, amide, amine salt and aldehyde groups. From the GC-MS analysis of TLC fraction of ethyl acetate extract, five different bioactive compounds e.g., 2,4-ditert -butylphenyl 5-hydroxypentanoate, Oxalic acid; allyl nonyl ester, 7,9-Ditert-butyl-1-oxaspiro(4,5)deca-6,9-diene- 2,8-dione, Dibutyl phthalate and 2,3,5,8-tetramethyl-decane were identified. Leaf extracts of P. pudica contain bioactive compounds that can be used as broad spectrum bactericidal agent.

Biological activities of in vitro liverwort Marchantia polymorpha L. extracts

To overcome the problems in liverwort collecting such as small size and easily mixed with other species in the wild, we have successfully cultivated Marchantia polymorpha L. under in vitro conditions in the previous study. The aim of this study is to evaluate the biological activities of this in vitro biomass as a confirmation of the sufficient protocol in cultivation this species. Cultured biomass was dried at a temperature of 45-50 oC to constant weight and ground into a fine powder. The coarse powder was extracted with organic solvents of increasing polarization including n-hexane, chloroform, ethyl acetate, and ethanol using the maceration technique. Four extracts were investigated antioxidant (iron reduction power, DPPH), antibacterial (agar diffusion), tyrosinase inhibitory activity, anti-proliferation on MCF-7 cells. Additionally, the presence of natural metabolite groups of the extracts was detected by using specific reagents. For antioxidant activity, ethyl acetate fraction extract had the highest iron reducing power and DPPH free radical scavenging ability with IC50 = 439.31 µg ml-1. All three n-hexane, chloroform, and ethyl acetate extracts possessed resistance to the bacterial strain tested. At a concentration of 2 mg ml-1, n-hexane and chloroform extracts had the highest percentage of tyrosinase inhibition (69.54 and 69.10%, respectively). The n-hexane extract is a potent extract that inhibits the proliferation of MCF-7 cells with the lowest IC50 of 38.15 µg ml-1. A preliminary chemical composition survey showed that the cultured biomass liverwort contains many bioactive compounds, particularly the compounds of range of non- and less-polarized fractions.

Comparative antioxidant and antimicrobial potentials of leaf successive extract fractions of poison bulb, Crinum asiaticum L.

Crinum asiaticum L. (family: Amaryllidaceae) is a widely used medicinal plant species having great ethnomedicinal values. The present study aimed to evaluate in vitro antioxidant and antimicrobial potentials of the leaf extracts of C. asiaticum with their phenolics content. The total antioxidant assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, and ferric ion reducing antioxidant power (FRAP) assays of the leaf crude aqueous extract and the five successive extract fractions were analyzed to evaluate their antioxidant potential. The antimicrobial potential was investigated against two gram-negative and one gram-positive bacteria strain by the disc diffusion method. The aqueous fraction of C. asiaticum (CaAq) showed relatively higher antimicrobial potentials than the crude aqueous extract (CaLAE), in terms of zone of inhibition, indicating the fact that serial extraction procedure can more successfully concentrate the specific bioactive components. Along with CaAq and CaLAE, CaEA fraction (ethyl acetate extract fraction) also exhibits antimicrobial activities probably due to the presence of terpenoids, phenolics, and flavonoids. It might be concluded that antioxidants and antimicrobial agents are available in the leaves of C. asiaticum where water and ethyl acetate might be the effective solvent systems to extract antimicrobial and antioxidant agents respectively.

Bioassay-Guided Fractionation of Endophytic Fungal Extract of Fusarium solani (Saccardo) against Cancer Cell Lines and Zebrafish Embryo

Abstract: Background: This study aimed to explore the toxicity and apoptotic potential of Fusarium solani extracts on cancer cell lines and zebrafish embryo. Materials and Methods: F. solani was initially extracted with ethyl acetate and methanol. Further purification was performed using column chromatography. Each fraction was assayed for its cytotoxic potential against four cancer cell lines using MTT assay. The apoptotic potential of the HepG2 cells was investigated using Hoechst 33342 dye and caspase 3/7. The in vivo developmental toxicity of the active fractions was assessed using zebrafish embryos. Results: The ethyl acetate extract and active fractions affected the cell viability of tested cell lines on dose dependent manner. The IC50 values of F1 fraction for Jurkat, HEK 293, MDA-MB-231 and HepG2 cell lines were 48.07, 60.09, 97. 82 and 69.7 μg/ml respectively whereas, the IC50 values of F7 fraction were 58.17μg/ml and 34.29 for HepG2 and HEK 293 cell lines respectively. Morphological changes of cell such as chromatin condensation as well as the expression of caspase 3/7 were observed. In agreement with cell line data, the active fraction obtained from F. solani were toxic for zebrafish embryos at ≥100 μg/ml and lower concentration (≤60μg/ml) induced various developmental defects. Conclusion: The F. solani ethyl acetate extract and its fractions showed anticancer activity in tested cancer cell lines and induced apoptosis via caspase 3/7 pathway and showed in vivo toxicity in zebrafish embryos. Key words: Cell lines, Endophyte, Fusarium solani, Toxicity, Zebrafish.

Macrophylloflavone: A New Biflavonoid from Garcinia macrophylla Mart. (Clusiaceae) for Antibacterial, Antioxidant, and Anti-Type 2 Diabetes Mellitus Activities.

Investigations of antibacterial, antioxidant, and anti-type 2 diabetes mellitus activities have been carried out on Garcinia macrophylla Mart. plant extract fractions. An isolate from a fraction of ethyl acetate extract was characterized with spectroscopic data. A new biflavonoid compound was found to have a skeleton of 5,7,4′,5″,7″,3‴,4‴-heptahydroxyflavanone[3-6″] flavones which was named macrophylloflavone (1). The compound was evaluated for its antibacterial activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 with cephazolin as a positive control, antioxidant assay against 2,2 diphenyl-1-picrylhydrazyl (DPPH) with ascorbic acid as the positive control, and anti-type 2 diabetes mellitus treatment with metformin as a positive control. The biflavonoid compound exhibited a good inhibition for bacteria and free radical DPPH. Furthermore, biflavonoid compound treatment on the diabetic rats suggested its ability to decrease the blood glucose level. This study provided evidence that the plant has antibacterial, antioxidant, and antidiabetic properties.

Comparative assessment of antimicrobial and antioxidant activity between whole plant and parts of Sphaeranthus\xa0indicus Linn. (Asteraceae)

BackgroundSphaeranthus indicus is an important medicinal plant, which is used to cure various illnesses. The present study is the first investigation of the antimicrobial, antioxidant and phytochemical analysis of Sphaeranthus indicus from Chhattisgarh, India.MethodsThe whole plant and plant parts were extracted with polar and non-polar solvents such as water, methanol, ethyl acetate and hexane to assess various bioactivities. The antimicrobial and antioxidant activities were performed by ager well diffusion method and ferrous reducing capacity, respectively; However free radical scavenging activity was analyzed using DMPD and DPPH scavenging assay. The DMPD and DPPH assay were performed in a time dependent manner. Qualitative and quantitative analysis were performed for the total phytochemicals present in the plant extracts. The total content of phenols, flavonoids and terpenoids was analyzed by colorimetric methods.ResultsEthyl acetate and hexane extract of plant inflorescence and stem exhibited significant antibacterial activity against tested bacterial pathogens. The clinically isolated gram positive pathogenic bacteria responded better as compared to clinically isolated gram negative bacteria as well as pathogenic gram positive and negative bacteria acquired from Microbial Type Culture Collection, India. The leaf and inflorescence exhibited potent antioxidant activity. The polar fraction of leaf methanol extract exhibited the highest reducing power capacity. The aqueous extract of inflorescence exhibited highest inhibition against DMPD and DPPH radicals. The whole plant aqueous extract showed maximum inhibition while aqueous extract of inflorescence exhibited high inhibition among different plant parts. Qualitative phytochemical analysis revealed the presence of terpenoids, phenols, flavonoids, tannins and cardiac glycosides in plant parts. Total terpenoid content was found to be highest in polar fraction of stem methanol extract and ethyl acetate extract of inflorescence. However, total phenol was found to be highest in polar fraction of leaf methanol extract, similarly highest flavonoid content was observed in aqueous extract of leaf.ConclusionThe results suggest that biological activities of plant parts depend on content of active phytochemicals. The inflorescence could be a potential source of antimicrobial and antioxidant compound. Further, investigation pertaining isolation and characterization of active ingredient may provide an insight regarding its phytochemical activity.

The Antioxidant Activity and Phytochemical Analysis of Medicinal Plant Veronica Biloba

A medicinal plant veronica genus has 450 well known species and found across both temperate and hemisphere region their 26 species are endemic and known in a total of 79 popular species and are widely utilized throughout the world due to important biological activities. In this study fully powdered uniform size specie veronica biloba plant taken in porous bag were manually subjected to soxhlet hot continuous process for cyclization of extraction using ethanol (300 mL) a concentrated dried extract obtained after solvent evaporation. Furthermore, liquid-liquid extracted fractions as water, dichloromethane, n-hexane, and ethyl acetate yields results founds polar fraction with highest percentage (water 47.51 %). The phytochemical screening of veronica biloba has shown all major compounds entirely present in extracts. One of the primitive phenolic compound flavonoid is present in plant and show potency towards antioxidants. All the extracted fractions of plant showed excellent antioxidant activity using a stable DPPH (1,1-diphenyl-2-picryl-hydrazyl) method at concentration (range from 31.25 to 500µg/mL). Primary our ethyl acetate extract fraction showed highest inhibition potential at IC50 = 1.70±0.05µg/mL which is much closer to a standard positive control Propyl gallate showed IC50 = 1.6±0.05µg/mL percent potential. The purification and isolation of these extract is important which can provide us help in novel antioxidants discovery also natural antioxidants currently in cosmetics products, food and therapeutics health related products significantly demanded because they are very effective, efficient and harmless as compared to synthetic one.

Bioactive Compounds in Moringa oleifera Lam. Leaves Inhibit the Pro-Inflammatory Mediators in Lipopolysaccharide-Induced Human Monocyte-Derived Macrophages.

Moringa oleifera (MO) is an important plant for traditional medicine. The present study aimed to identify the MO active phytochemical compounds for their ability against inflamed macrophages. An ethyl acetate extract fraction of MO was fractionation by flash column chromatography. Human macrophages were stimulated by Lipopolysaccharide and then treated with fractions of MO to examine their anti-inflammatory activity and cellular mechanism. The active fractions were analyzed by liquid chromatography with electrospray ionization quadrupole time-of-flight mass spectrometer (LC-ESI-QTOF-MS). MO treated cells showed a decreased production of pro-inflammatory mediator in response to lipopolysaccharide. This was evident at both mRNA and protein levels. The study revealed that MO suppressed mRNA expression of IL-1, IL-6, TNF-α, PTGS2, NF-κB (P50), and RelA. Furthermore, the extract effectively inhibited the expression of inflammatory mediators, including IL-6, TNF-α, and cyclooxygenase-2. Interestingly, the effect of MO inhibited phosphorylation of IκB-α and the ability to reduce expression of the nuclear factor (NF)-κB p65, suppressing its nuclear translocation. Moreover, LC-ESI-QTOF-MS analysis of the MO active fraction revealed seven compounds, namely 3,4-Methyleneazelaic acid, (2S)-2-phenylmethoxybutane-1,4-diol, (2R)-2-phenylmethoxybutane-1, 4-diol, γ-Diosphenol, 2,2,4,4-Tetramethyl-6-(1-oxobutyl)-1,3,5-cyclohexanetrione, 3-Hydroxy-β-ionone, and Tuberonic acid. Our findings highlight the ability of MO compounds to inhibit inflammation through regulation of the NF-κB pathway.

The antioxidant activity and phytochemical analysis of medicinal plant Veronica biloba

A medicinal plant veronica genus has 450 well known species and found across both temperate and hemisphere region their 26 species are endemic and known in a total of 79 popular species and are widely utilized throughout the world due to important biological activities. In this study fully powdered uniform size specie veronica biloba plant taken in porous bag were manually subjected to soxhlet hot continuous process for cyclization of extraction using ethanol (300 mL) a concentrated dried extract obtained after solvent evaporation. Furthermore, liquid-liquid extracted fractions as water, dichloromethane, n-hexane, and ethyl acetate yields results found polar fraction with the highest percentage (water 47.51 %). The phytochemical screening of veronica biloba has shown all major compounds entirely present in extracts. One of the primitive phenolic compound flavonoid is present in plant and show potency towards antioxidants. All the extracted fractions of plant showed excellent antioxidant activity using a stable DPPH (1,1-diphenyl-2-picryl-hydrazyl) method at concentration (range from 31.25 to 500µg/mL). Primary our ethyl acetate extract fraction showed the highest inhibition potential at IC50 = 1.70±0.05µg/mL which is much closer to a standard positive control Propyl gallate showed IC50 = 1.6±0.05µg/mL percent potential. The purification and isolation of these extract areimportant which can provide us help in novel antioxidants discovery also natural antioxidants currently in cosmetics products, food and therapeutics health related products significantly demanded because they are very effective, efficient and harmless as compared to synthetic.

Bioactive pterocarpans from Trigonella foenum-graecum L

Trigonella foenum-graecum L. (fenugreek) is used as a leafy vegetable and spice in China and North African countries. However, the biochemical components of its aerial parts were rarely explored. In this study, the bioactivities of the various extract fractions from the aerial parts of this edible plant were assessed, the ethyl acetate extract fraction exhibited strong antioxidant and anti-inflammatory effects. Through bioassay-guided isolation, one new pterocarpan (1), as well as twelve known pterocarpans (2–13) were obtained, nine of them (5–13) were first reported in the fenugreek, four pterocarpans (9, 11–13) had strong antioxidant activity, eleven pterocarpans (1–3, 5–12) possessed obvious anti-inflammatory activity. This study indicates that pterocarpans are main bioactive components of this edible plant. Apart from its nutritional value as food, the aerial parts of this plant can also be further explored as functional foods or antioxidants in food industry.

Uji Antimikroba Fraksi Ekstrak Metanol, Etil Asetat dan n-Heksana Daun Tabar-Tabar (Costus speciosus) dan Toksisitasnya Terhadap Larva Udang

A Study on antimicrobial examination of fractions of methanol, ethyl-acetate, and n-hexane extract of tabar-tabar (Costus speciosus) leaf to bacteria (Bacillus sp., Staphylococcus aureus, Escherichia coli, and Serratia marcescens) and yeast (Candida albicans), and their toxicity to brine-shrimp (Artemia salina) larvae has been done. Examination of antimicrobial activities to the microbes were done by agar diffusion method, while test for toxicity was done by exposing 2 days-old larvae to certain concentration of the extract. Preliminary tests on phytochemical compounds in leaf were done by Mayer’s, FeCl3, Mg-HCl, foam, and 10% NaOH in methanol tests. The results showed that methanol leaf extract of tabar-tabar in general inhibited more in the growth of gram-positive Bacillus sp. and S. aureus rather than to other tested microbes. Compared to methanol and ethyl-acetate fraction, n-hexane fraction of the leaf inhibited more Bacillus sp. and E. coli. Interestingly, C. albicans was highly inhibited by n-hexane fractions. LC50 of methanol extract, ethyl-acetate, and n-hexane fraction, were 45.53, 478.37, and 824.20 ppm, respectively. Preliminary test on phytochemical compounds showed saponin was only detected in fractions of methanol and ethyl acetate extracts, steroid was detected in fraction of methanol, while triterpenoid was detected in fraction of n-hexane extract. Phenolic was detected in all extract fractions, while flavonoid, kumarin, and alkaloid on the other hand were not detected.

Ethyl acetate fraction of Psorospermum febrifugum Spach aqueous extract did not exhibit acute or sub-chronic toxicity. Experimental study on Wistar rats

Psorospermum febrifugum Spach (Clusiaceae) was a tropical plant whose root bark was used to treat anemia. This work aimed to evaluate the safety of the ethyl acetate fraction of the aqueous extract of this bark. Methods: The ethyl acetate fraction of the extract was administered to Wistar rats in a single dose of 2000 mg / Kg body weight for acute oral toxicity test or daily doses of 200 mg / Kg of body weight during 28 days for sub-chronic oral toxicity test, as recommended by the OECD. At day 0, then at day 14 for the acute phase and day 28 for the sub-chronic phase, the rats were weighed and their blood collected for tests. The activity of transaminases AST and ALT were measured in the liver function tests, blood urea and creatinine were measured for renal function tests and blood leukocytes were counted for the immune balance. These analyzes were supplemented by the histology of the liver, kidneys and spleen, an immune organ. Results: In acute and sub-chronic oral toxicity tests, rat’s weight, liver, kidney and immune balances as well as these organs histology were not affected, suggesting the safety of the extract fraction. Conclusion: The ethyl acetate fraction of the aqueous extract of the root bark of Psorospermum febrifugum did not reveal any acute or sub-chronic oral toxicity. This effect could be related to its richness in flavonoids which have cytoprotective effects. The study of biological tolerance deserves to be continued by the chronic toxicity test and appropriate clinical trials.

Antioxidant Activity of Daemonorops draco Resin

Jernang resin is secretion of jernang rattan (Daemonorops draco, Arecaceae family) fruits which is endemic in Southeast Asia. This resin has various biological activities and empirically used as wound healing, headache medicines, and fever remedies by Anak Dalam ethnic group from Jambi. This study was performed to evaluate the antioxidant activity of nonpolar fraction of D. draco resin which collected from Jambi Province, Sumatera, Indonesia. Resin was extracted with n-hexane, ethyl acetate, and methanol respectively. The antioxidant properties of the extracts were then evaluated using 1,1-diphenyl-2picryl-hidrazyl radical scavenging assay. The most active extract was further fractionated using n-hexane and methanol and separated using column chromatography and preparative thin layer chromatography. Separation of the extract was conducted through antioxidant assay-guided fractionation. Characterization of the active fraction was carried out by infrared spectroscopy. The result shows that ethyl acetate extract provides higher antioxidant activity (IC50 = 27.61 µg/mL) compare to methanol and n-hexane extracts. N-hexane fraction of ethyl acetate extract used for further separation using column and preparative thin layer chromatography due to its antioxidant activity. Separation using column chromatography resulting in 9 fractions (F.1-9). Fraction F.5 provide high antioxidant activity (IC50 = 17.27 µg/mL) and further separated using preparative thin layer chromatography resulting two fractions with lower antioxidant activity F.5.1 (IC50 = 85.18 µg/mL) and F.5.2 (IC50 = 34.94 µg/mL). Characterization of fraction F.5.2 using infrared spectroscopy showed that component in fraction F.5.2 contains NH-substituted benzene.

Active Constituent in the Ethyl Acetate Extract Fraction of Terminalia bellirica Fruit Exhibits Antioxidation, Antifibrosis, and Proapoptosis Capabilities In Vitro.

Terminalia bellirica (Gaertn.) Roxb. fruit (TBF) is a widely planted traditional medicinal herb in Tibet. We aimed to determine the most active substance-enriched extract by comparing the in vitro antioxidant activities of different extract fractions of TBF that were subsequently extracted by petroleum ether, chloroform, ethyl acetate, and n-butanol after initial extraction by 95% ethanol. The main compounds of the ethyl acetate extract fraction (EF) were analyzed via HPLC-MS. Gallic acid (GA) was obtained from EF to determine in vitro antifibrotic activity based on the traditional usage of TBF. After HSC-T6 cells were incubated with GA, extracellular secreted levels of fibrosis-associated cytokines, such as collagen I, collagen III, TGF-β1, and hydroxyproline, were estimated by ELISA. Gene and protein expressions of PDGFR, CTGF, NF-κB, MMP-2, TIMP-1, TIMP-2, α-SMA, and the Bcl-2/Bax family were determined by quantitative PCR and western blot. The proapoptotic effect of GA was further investigated by annexin V-PI and TUNEL staining. These results indicate that EF has prominent in vitro antioxidant activity among four extract fractions, and its main component, GA, manifests antifibrosis activity and its potential mechanism of action includes inhibition of cytokine secretion and collagen synthesis, as well as proapoptosis of HSCs.

Sub-chronic oral toxicity profile of Carpolobia lutea leaf fractions in rats

Carpolobia lutea (CL) (Polygalaceae) leaf is widely reported to be effective for the treatment of diarrheal diseases. The subchronic oral toxicological investigation is yet to be executed with both the crude ethyl acetate extract (CEAE) and ethyl acetate fraction (EAF) to verify suitability of plant for the management of diarrhea diseases. The pathological alteration following oral administration of CEAE and EAF of C. lutea leaf were explored in rodents over a period of 60 days adopting Good Laboratory Practice (GLP) conditions. Thirty adult male and female wistar rats were randomized to four treatment groups of 6 rats /sex/group and were administered orally with three doses of CEAE, 192.5, 385.0, 770 mg / kg and one dose of EAF 770.0 mg/kg; while 10ml/kg of 20% Tween 80 was used for the control test. Weekly relative body weight and vital signs were assessed. Blood samples were collected weekly for hematological assessment. At autopsy, the major organs were cautiously excised and weighed. The study showed that both CEAE and EAF do not have significant impact (P > 0.05) on the hematopoietic system rather it impact more on the biochemical parameters which increased significantly (P < 0.05 - 0.001) and dose dependently. These results indicate that oral administration of CEAE and EAF in rats mediates biochemical but not haematopoietic sub-chronic toxicity. Its pharmacological and therapeutic effectiveness is not without toxicity implication and should be used cautiously sub-chronically.