Bioassay-guided Fractionation Of Ethanol Extract Research Articles

Scientific Baseline Information for the Potential Use of Hibiscus surattensis L against Malaria: Phytochemistry and Biological Studies

Background and aim: Hibiscus surattensis L. is a medicinal plant widely used traditionally in Benin to treat malaria. This study was designed to investigate antiplasmodial activity, hemolytic power, acute toxicity, antioxidant activity and phytochemical content of H. surattensis.
 Methodology: Bioassay-guided isolation approach was adopted and extracts and fractions collected were continuously tested in vitro against Chloroquine-sensitive and field isolate strains of Plasmodium falciparum by immune-dosage of Plasmodium lactate dehydrogenase. Hemolytic effect and acute toxicity of extracts were evaluated respectively on human erythrocytes and according to OECD guideline N°423. 2, 2-diphenyl-1- picrylhydrazyl) radical scavenging, Ferric reducing antioxidant power, superoxyde radical scavenging and hydrogen peroxide radical scavenging methods were used to investigate antioxidant activity.
 Results and conclusion: The best antiplasmodial activity was obtained with ethanolic extract of H. surattensis L. against field isolate (IC50 = 5.03±0.31 µg/mL) and Chloroquine -sensitive (IC50 = 7.55±0.59 µg/mL) whereas aqueous extract exhibited moderate activity. Bioassay-guided fractionation of ethanolic extract shows progressive decrease of the antiplasmodial activity. Both extracts exhibited strong antioxidant activity, hemolytic power less than 1%. No mortality of rats was recorded with ethanolic extract at 2000 mg/kg body weight. Flavonoids, anthraquinones, coumarins, and triterpenes are present in both extracts with tannins in the ethanolic extract. In summary, the extracts of H. surattensis have interesting antiplasmodial and antioxidant properties probably resulting from a synergetic action of their secondary metabolites, without toxicity effect on rats and human erythrocytes. These findings strengthen the traditional use of H. surattensis as antimalarial plants.
 Keywords: Hibiscus surattensis; antiplasmodial; antioxidant; toxicity; phytoconstituents.

Antiproliferative activity of Haematoxylum brasiletto H. Karst.

Background:Haematoxylum brasiletto is a tree that grows in Central America, commonly known as “Palo de Brasil,” which is used in the traditional medicine for the treatment of cancer and gastric ulcers.Objective:The aim of this study was to isolate the compounds responsible for antiproliferative activity of H. brasiletto.Materials and Methods:A bioassay-guided fractionation of ethanol extract of H. brasiletto was performed using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide cell proliferation assay to measure the antiproliferative activity on six human cancer cell lines (A549, LS180, HeLa, SiHa, MDA-MB-231, and NCI-H1299) and one human noncancer cell line (ARPE-19). The ethanol extract was partitioned with hexane, dichloromethane, and ethyl acetate. The active dichloromethane fraction was fractioned by silica-column chromatography, and active subfractions were separated using preparative-thin layer chromatography. The chemical structure of an isolated compound was elucidated with different chemical and spectroscopic methods.Results:The flavonoid brazilin (1) was isolated from the heartwood of H. brasiletto. The measurement of antiproliferative activity showed that brazilin can inhibit the growth of SiHa, MDA-MB-231, A549, and NCI-H1299 cell lines by 50% at doses of 44.3, 48.7, 45.4, and 48.7 μM, respectively. Furthermore, the flavonoid showed a high antiproliferative activity on LS 180 and HeLa with IC50 values of 62.2 and 71.9 μM, respectively. Brazilin also exhibited a high antiproliferative activity on the human noncancer cell line ARPE-19 with an IC50 value of 37.9 μM.Conclusions:Brazilin: (6aS, 11bR)-7,11b-Dihidro-6H-indeno[2,1-c] cromeno-3,6a, 9,10-tetrol was isolated; this compound demonstrated antiproliferative activity against several human cancer cell lines. This work demonstrated that brazilin, a flavonoid isolated and characterized of H. brasiletto, has antiproliferative activity against cancer cell lines.SUMMARY The flavonoid brazilin was isolated from the heartwood of H. brasilettoBrazilin is able to inhibit the growth of SiHa, MDA-MB-231, A549 and NCI- H1299 cancerous cell linesBrazilin exhibited a moderate antiproliferative activity on the human non-cancer cell line ARPE-19Brazilin demonstrated to have antiproliferative activity against human cancer cell lines and could be a potential source of anticancer agents. Abbreviations used: MTT: [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium]; FBS: Fetal bovine serum; TLC: Thin layer chromatography.

Anti-quorum sensing activity of flavonoid-rich fraction from Centella asiatica L. against Pseudomonas aeruginosa PAO1.

Inhibition of quorum sensing (QS), a cell-density dependent regulation of gene expression in bacteria by autoinducers is an attractive strategy for the development of antipathogenic agents. In this study, the anti-QS activity of the ethanolic extract of the traditional herb Centella asiatica was investigated by the biosensor bioassay using Chromobacterium violaceum CV026. The effect of ethyl acetate fraction (CEA) from the bioassay-guided fractionation of ethanol extract on QS-regulated violacein production in C. violaceum ATCC12472 and pyocyanin production, proteolytic and elastolytic activities, swarming motility, and biofilm formation in Pseudomonas aeruginosa PAO1 were evaluated. Possible mechanism of QS-inhibitory action on autoinducer activity was determined by measuring the acyl homoserine lactone using C. violaceum ATCC31532. Anti-QS compounds in the CEA fraction were identified using thin layer chromatography biosensor overlay assay. Ethanol extract of C. asiatica showed QS inhibition in C. violaceum CV026. Bioassay-guided fractionation of ethanol extract revealed that CEA was four times more active than the ethanol extract. CEA, at 400μg/mL, completely inhibited violacein production in C. violaceum ATCC12472 without significantly affecting growth. CEA also showed inhibition of QS-regulated phenotypes, namely, pyocyanin production, elastolytic and proteolytic activities, swarming motility, and biofilm formation in P. aeruginosa PAO1 in a concentration-dependent manner. Thin layer chromatography of CEA with biosensor overlay showed anti-QS spot with an Rf value that corresponded with that of standard kaempferol. The anti-QS nature of C. asiatica herb can be further exploited for the formulation of drugs targeting bacterial infections where pathogenicity is mediated through QS.

Antibacterial activities and cytotoxicity of terpenoids isolated from Spirostachys africana

Spirostachys africana Sond. stem bark is used traditionally for the treatment of diarrhoea and dysentery in Limpopo Province of South Africa. Bioassay-guided fractionation of ethanolic extract from bark of Spirostachys africana led to the isolation of four known compounds, two triterpenoids, compound 1 [ d-Friedoolean-14-en-oic acid (3-acetyl aleuritolic acid)] and compound 2 (Lupeol), and two diterpenes, compound 3 [ ent-2,6α-dihydroxy-norbeyer-1,4,15-trien-3-one (diosphenol 2)] and compound 4 ( ent-3β-hydroxy-beyer-15-ene-2-one). Isolated compounds were tested for antibacterial activity using micro-dilution method. Compound 1, exhibited minimum inhibitory concentration (MIC) of 50 μg/ml against Staphylococcus aureus, Salmonella typhy, Vibrio cholera, Escherichia coli and Shigella dysentery. Compound 2 was not active against all tested microorganisms at 200 μg/ml, which was the highest concentration tested. At this concentration, all four compounds were not active against Shigella sonnei. Cytotoxicity of ethanol crude extracts and isolated compounds from Spirostachys africana was determined using the sodium-2,3-bis-[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) assay on Vero cells. Compounds 2 and 3, isolated from Spirostachys africana, had up to three times higher [50% inhibitory concentration (IC 50 values; 300.9 and 308.9 μg/ml)] than the ethanol crude extracts (102.8 μg/ml) suggesting higher toxicity of the crude extract as compared to these two compounds. In contrast, compounds 1 and 4 were not cytotoxic to Vero cell lines (African green monkey) in vitro at the concentrations tested (IC 50 > 400 μg/ml). This is the first report on the antibacterial activity and cytotoxicity of purified compounds from Spirostachys africana.

Sesquiterpenes from the wood of Juniperus lucayana

Bioassay-guided fractionation of ethanolic extract from the wood of Juniperus lucayana afforded three sesquiterpenes named 3-hydroxypseudowiddran-6(7)-en-4-ol ( 1), 15-hydroxyallo-cedrol ( 2) and 12-hydroxywiddrol ( 3) together with six known sesquiterpenes ( 4– 9) and two known flavonoids ( 10 and 11). Their structures were established on the basis of comprehensive spectroscopic analyses, including 2D NMR spectroscopy and mass spectrometry. The structures of compounds were identified as 1α,4β,11α,11β-tetramethylbicyclo[5,4,0]undec-6(7)-en-3α, 4α-diol ( 1), 4β-hydroxymethyl-5,5,9β-trimethyltricyclo[4.3.0.2 1.4]undecan-3α-ol ( 2) and 4β-hydroxymethyl-7α,11α,11β-trimethylbicyclo [5.4.0]undec-1-en-4α-ol ( 3). The major compounds isolated were evaluated for their antifungal activity against Botrytis cinerea. Widdrol ( 7) was the most active, reaching the 71% inhibition level on mycelial growth after 6 days.