A comparative study of in vitro antimicrobial, antioxidant and cytotoxic activity of Albizia lebbeck and Acacia nilotica stem bark

This study aims to explore the cytotoxic test (LC 50 ) towards Artemia salina L. larvae of the partition ethyl acetate fractions from methanol extracts and petroleum ether (PE) extracts of Pistiae folium . The leaves of Pistiae folium was extracted using solvents soxhletation with methanol solvent and petroleum ether solvent. Both extracts successively partitioned with n-hexane and ethyl acetate, respectively. Identification of secondary metabolites was using phytochemical screening method and TLC (Thin Layer Chromatography). TLC analysis was using the eluent chloroform: methanol 3: 1 by citroboric acid spot analysis. The cytotoxic activity was using Brine Shrimp Lethality Test (BSLT) method. The result of partition from methanol extracts was 6.25% and from PE extracts was 0.69%. The phytochemical screening test showed that it contained flavonoids, saponins and steroids. The TLC analysis identified flavonoid compounds. The cytotoxic activity (LC 50 ) of the ethyl acetate partition from methanol extracts and petroleum ether extracts of Pistiae folium were 79.9298mg/mL and 51.7608mg/mL, respectively. The result showed that the cytotoxic activity of ethyl acetate fractions partitined from methanol extract of Pistiae folium was higher than the PE extracts.

BackgroundThe use of plants and their derived substances increases day by day for the discovery of therapeutic agents owing to their versatile applications. Current research is directed towards finding naturally-occurring antioxidants having anticancer properties from plant origin since oxidants play a crucial role in developing various human diseases. The present study was designed to investigate the antioxidant and anticancer properties of Sygygium fruticosum (Roxb.) (abbreviated as SF).MethodsThe dried coarse powder of seeds of SF was exhaustively extracted with methanol and the resulting crude methanolic extract (CME) was successively fractionated with petroleum ether, chloroform and ethyl acetate to get petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and lastly aqueous (AQF) fraction. The antioxidant activities were determined by several assays: total antioxidant capacity assay, DPPH free radical scavenging assay, hydroxyl radical scavenging assay, ferrous reducing antioxidant capacity and lipid peroxidation inhibition assay. The in vivo anticancer activity of SF was determined on Ehrlich’s Ascite cell (EAC) induced Swiss albino mice.ResultsAll the extractives showed strong antioxidant activities related to the standard. The total antioxidant capacity (TAC) of the fractions was in the following order: EAF>AQF>CME>PEF>CHF. The TAC of EAF at 320 μg/mL was 2.60±0.005 which was significantly higher (p < 0.01) than that of standard catechin (1.37 ± 0.005). The ferrous reducing antioxidant capacity of the extracts was in the following order: EAF>AQF>CME>AA>CHF>PEF. In DPPH free radical scavenging assay, the IC50 value of EAF was 4.85 μg/mL, whereas that of BHT was 9.85 μg/mL. In hydroxyl radical scavenging assay and lipid peroxidation inhibition assay, the EAF showed the most potent inhibitory activity with IC50 of 43.3 and 68.11 μg/mL, respectively. The lipid peroxidation inhibition assay was positively correlated (p < 0 .001) with both DPPH free radical scavenging and hydroxyl radical scavenging assay. The total phenolic contents of SF were also positively correlated (p < 0 .001) with DPPH free radical scavenging, hydroxyl radical scavenging and lipid peroxidation inhibition assay. Based on antioxidant activity, EAF was selected for cytotoxic assay and it was found that EAF inhibited 67.36% (p < 0.01) cell growth at a dose of 50 mg/kg (ip) on day six of EAC cell incubation.ConclusionsOur results suggest that EAF of seeds of SF possess significant antioxidant and moderate anticancer properties. Seeds of SF may therefore be a good source for natural antioxidants and a possible pharmaceutical supplement.

Antioxidant activity of petroleum ether, chloroform, ethyl acetate and methanol extracts of leaf, stem and root of Mimosa pudica L. was observed through DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay. Five concentrations (12.5, 25.0, 50.0, 100.0 and 200.0µg/ml) were taken for each extract as well as the standard and the absorbances were measured at 517nm using a spectrophotometer against methanol blank. The activity was increased by the increment of concentrations of the extracts. In case of leaf, the highest scavenging percentage was found in chloroform extract (86.40%) at 200.0µg/ml concentration. But for stem and root, the highest scavenging percentages were found in ethyl acetate extracts (73.72% and 83.79% respectively) at same concentration. The ethyl acetate extracts showed the highest activity among all the extracts where the IC50 values were 65.152µg/ml, 76.036µg/ml and 65.000µg/ml and the lowest was found in petroleum ether extracts where the IC50 values were 130.129µg/ml, 147.891µg/ml and 186.449µg/ml for leaf, stem and root respectively and that was for ascorbic acid (standard) was 18.012µg/ml.

Qualitative phytochemical and anti-trypanosomal properties of the petroleum ether, chloroform, methanol and aqueous extracts, obtained by cold extraction from the leaves, stem bark and roots of Prosopis africana were evaluated. The methanolic and aqueous extracts of the stem bark and leaves of the plant contained alkaloids, saponins, tannins and flavonoids. Anthraquinone was present in the stem bark methanolic extract and in the methanolic and aqueous extracts of the root as well as the aqueous extract of the leaves. Resins was present in the petroleum ether and chloroform extracts of the stem bark and leaves, while tannins was detected in the methanol and aqueous extracts of the stem bark and leaves of the plant in addition to the petroleum ether and chloroform extracts of the root bark. All the solvent extracts showed strong in vitro anti-trypanosomal activity and 2 and 4 mg/ml, but in vivo only the methanolic extract of the leaves displayed the most promising anti-trypanosomal effect at 200 mg/kg dose. Hence, Prosopis africana extracts possess significant anti-trypanosomal activity to warrant bioassay-guided evaluation and identification of the active principle.

Use of different solvent systems for extraction of plant materials may cause variation in their bioactivities. The present study was conducted to evaluate the presence of different phytoconstituents and to compare in vitro bioactivities of petroleum ether, dichloromethane (DCM) and methanol extracts of Bombax ceiba (B. ceiba) roots available in Bangladesh. Preliminary phytochemical screening was conducted using specific standard procedure. Antioxidant activity of the extracts was evaluated using DPPH radical scavenging assay. Determination of total phenolic and flavonoid content was also carried out. Antibacterial and cytotoxic activities were investigated using disc diffusion method and brine shrimp lethality bioassay, respectively. All the experiments were carried out from February 2016 to September 2016. Phytochemical evaluation revealed the presence of alkaloids, terpenoids, carbohydrates, tannins, flavonoids, saponins and steroids. The methanol extract showed the highest DPPH radical scavenging activity and had the highest phenolic (187.42 ± 3.77 mg/g, GAE) and flavonoid content (74.67 ± 4 mg/g, QE) followed by the DCM and petroleum ether extracts. The extracts showed positive correlation between DPPH radical scavenging activity with the phenolic and flavonoid content. All the extracts showed mild to moderate in vitro antibacterial activity with zone of inhibition ranging from 7 mm to 13 mm. In brine shrimp lethality bioassay, the observed LC50 values for petroleum ether, DCM and methanol extracts were 70.72 μg/ml, 37.72 μg/ml and 22.58 μg/ml, respectively which revealed strong cytotoxic potential of the extracts compared to the positive control. The results indicated that B. ceiba roots could be a very potent source of natural radical scavenger and cytotoxic agent.

Use of different solvent systems for extraction of plant materials may cause variation in their bioactivities. The present study was conducted to evaluate the presence of different phytoconstituents and to compare in vitro bioactivities of petroleum ether, dichloromethane (DCM) and methanol extracts of Bombax ceiba (B. ceiba) roots available in Bangladesh. Preliminary phytochemical screening was conducted using specific standard procedure. Antioxidant activity of the extracts was evaluated using DPPH radical scavenging assay. Determination of total phenolic and flavonoid content was also carried out. Antibacterial and cytotoxic activities were investigated using disc diffusion method and brine shrimp lethality bioassay, respectively. All the experiments were carried out from February 2016 to September 2016. Phytochemical evaluation revealed the presence of alkaloids, terpenoids, carbohydrates, tannins, flavonoids, saponins and steroids. The methanol extract showed the highest DPPH radical scavenging activity and had the highest phenolic (187.42 ± 3.77 mg/g, GAE) and flavonoid content (74.67 ± 4 mg/g, QE) followed by the DCM and petroleum ether extracts. The extracts showed positive correlation between DPPH radical scavenging activity with the phenolic and flavonoid content. All the extracts showed mild to moderate in vitro antibacterial activity with zone of inhibition ranging from 7 mm to 13 mm. In brine shrimp lethality bioassay, the observed LC50 values for petroleum ether, DCM and methanol extracts were 70.72 μg/ml, 37.72 μg/ml and 22.58 μg/ml, respectively which revealed strong cytotoxic potential of the extracts compared to the positive control. The results indicated that B. ceiba roots could be a very potent source of natural radical scavenger and cytotoxic agent.

BackgroundMangifera pajang Kosterm is a plant species from the mango family (Anacardiaceae). The fruits are edible and have been reported to have high antioxidant content. However, the detailed phytochemical studies of the plant have not been reported previously. This study investigates the phytochemicals and biological activities of different parts of Mangifera pajang.MethodsThe plant samples were extracted with solvents of different polarity to obtain the crude extracts. The isolated compounds were characterized using spectroscopic methods. The extracts and isolated compounds were subjected to cytotoxicity tests using human breast cancer (MCF-7), human cervical cancer (HeLa) and human colon cancer (HT-29) cells. The free radical scavenging activity test was conducted using the DPPH assay. Antimicrobial activity tests were carried out by using the disc diffusion method.ResultsPhytochemical investigation on the kernel, stem bark and leaves of Mangifera pajang led to the isolation of methyl gallate (1), mixture of benzaldehyde (2) and benzyl alcohol (3), mangiferonic acid (4), 3β-hydroxy-cycloart-24-ene-26-oic acid (5), 3β,23-dihydroxy-cycloart-24-ene-26-oic acid (6), lupeol(7) lupenone(8), β-sitosterol(9), stigmasterol(10), trans-sobrerol(11) and quercitrin (12). Crude ethyl acetate and methanol extracts from the kernel indicated strong cytotoxic activity towards MCF-7 and HeLa cells with IC50 values of less than 10 μg/mL, while petroleum ether, chloroform and ethyl acetate extracts of the stem bark showed strong to moderate activity against MCF-7, HeLa and HT-29 cancer cell lines with IC50 values ranging from 5 to 30 μg/mL. As for the antimicrobial assays, only the ethyl acetate and methanol extracts from the kernel displayed some inhibition against the microbes in the antibacterial assays. The kernel extracts showed highest free radical scavenging activity with IC50 values of less than 10 μg/mL, while the ethyl acetate and methanol extracts of leaves displayed only weak activity in the DPPH assays.ConclusionsPhytochemical investigations on various parts of Mangifera pajang have identified terpenoids and a flavonol derivative as major constituents. Bioassay studies have indicated that the crude extracts and isolated compounds have potential as naturally-derived anticancer and antimicrobial agents, besides possess high free radical scavenging activity.

Protective effects of Flacourtia indica aerial parts extracts against paracetamol-induced hepatotoxiciy in rats

BackgroundRumex species are traditionally used for the treatment of neurological disorders including headache, migraine, depression, paralysis etc. Several species have been scientifically validated for antioxidant and anticholinestrase potentials. This study aims to investigate Rumex hastatus D. Don crude methanolic extract, subsequent fractions, saponins and flavonoids for acetylcholinestrase, butyrylcholinestrase inhibition and diverse antioxidant activities to validate its folkloric uses in neurological disorders. Rumex hastatus crude methanolic extract (Rh. Cr), subsequent fractions; n-hexane (Rh. Hex), chloroform (Rh. Chf), ethyl acetate (Rh. EtAc), aqueous fraction (Rh. Aq), crude saponins (Rh. Sp) and flavonoids (Rh. Fl) were investigated against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) at various concentrations (125, 250, 500, 1000 μg/mL) using Ellman’s spectrophotometric analysis. Antioxidant potentials of Rh. Sp and Rh. Fl were evaluated using DPPH, H2O2 and ABTS free radical scavenging assays at 62.5, 125, 250, 500, 1000 μg/mL.ResultsAll the test samples showed concentration dependent cholinesterase inhibition and radicals scavenging activity. The AChE inhibition potential of Rh. Sp and Rh. Fl were most prominent i.e., 81.67 ± 0.88 and 91.62 ± 1.67 at highest concentration with IC50 135 and 20 μg/mL respectively. All the subsequent fractions exhibited moderate to high AChE inhibition i.e., Rh. Cr, Rh. Hex, Rh. Chf, Rh. EtAc and Rh. Aq showed IC50 218, 1420, 75, 115 and 1210 μg/mL respectively. Similarly, against BChE various plant extracts i.e., Rh. Sp, Rh. Fl, Rh. Cr, Rh. Hex, Rh. Chf, Rh. EtAc and Rh. Aq resulted IC50 165, 175, 265, 890, 92, 115 and 220 μg/mL respectively. In DPPH free radical scavenging assay, Rh. Sp and Rh. Fl showed comparable results with the positive control i.e., 63.34 ± 0.98 and 76.93 ± 1.13% scavenging at 1 mg/mL concentration (IC50 312 and 104 μg/mL) respectively. The percent ABTS radical scavenging potential exhibited by Rh. Sp and Rh. Fl (1000 μg/mL) were 82.58 ± 0.52 and 88.25 ± 0.67 with IC50 18 and 9 μg/mL respectively. Similarly in H2O2 scavenging assay, the Rh. Sp and Rh. Fl exhibited IC50 175 and 275 μg/mL respectively.ConclusionThe strong anticholinesterase and antioxidant activities of Rh. Sp, Rh. Fl and various fractions of R. hastatus support the purported ethnomedicinal uses and recommend R. hastatus as a possible remedy for the treatment of AD and neurodegenerative disorders.

The extracts from the stem bark of Commiphora pedunculata, a plant used in Northern Nigeria for the treatment of infectious diseases, were subjected to phytochemical as well as antimicrobial screening using standard procedures. The antimicrobial activity against S. aureus, B. cereus, S. typhii, E. coli and C. albicans was carried out using the disc diffusion and broth micro dilution methods as outlined by the NCCLS. Preliminary phytochemical screening of the petroleum ether, ethyl acetate and methanol extracts revealed the presence of cardiac glycosides, anthraquinones, saponins, triterpenes, steroids, flavonoids, tannins and alkaloids. The results of the antimicrobial activity as indicated by the zone of inhibition of growth of the test microorganisms ranged from 17 to 28 mm, the MIC results ranged from 3.125 to 12.5 mg/mL and the MBC results ranged from 6.25 to 25.0 mg/mL for the petroleum ether, ethyl acetate and methanol extracts. The MIC of 12.50 mg/mL exhibited by the petroleum ether extract against both Gram-positive and Gram-negative bacteria indicates broad spectrum activity of Commiphora pedunculata. The results from this study showed that the extracts from the stem bark of the plant contain antimicrobial components worthy of further investigation and lends credence to the use of the plant for the treatment of infectious diseases.Keywords: Phytochemistry, Commiphora pedunculata extracts, antibacterial activity, MIC, MBC.

The methanol, petroleum ether, chloroform, ethyl acetate, n-butanol and water extracts were obtained by extraction of marigold flower (Calendula officinalis L). The content of total phenolic compounds, determined by UV spectrophotometric method using the Folin-Ciocalteu reagent, was 15.12 mg/g. The content of total flavonoids, determined by UV spectrophotometric method according to Markham, was 5.13 mg/g. Qualitative determination of phenolic compounds in the extracts was performed by one- and two-dimensional thin-layer chromatography (TLC) procedures. The results of one- and two-dimensional TLC analyses showed that different flavonoids and phenolic acids were present in the investigated extracts. The greatest number of flavonoids (rutin, quercetin and some unidentified flavonoid glycosides) and phenolic acids (chlorogenic, caffeic, coumaric and vanillic acid) were deteminated in methanol extract. The influence of marigold extracts, in concentration range 0.6-1.2 mg/mL, on 2,2?-diphenyl-1-picrylhydrazyl (DPPH) free radicals was investigated by electron spin resonance (ESR) spectroscopy. All extracts showed scavenging activity (SA) in the following order: ethyl acetate &gt; n-butanol &gt; methanol &gt; water &gt; chloroform &gt; petroleum ether. The SA increased with increasing concentration of extracts. The ethyl acetate and n-butanol extracts exibited the most significant SA. These extracts in concentration of 1.2 mg/mL eliminated completely DPPH radicals. The lowest SA had chloroform and petroleum ether extracts (in concentration of 0.6 mg/mL SA=0%). The SA of marigold extracts is attributed to its hydrogen-donating ability and scavenging effect.

: Objective: To determine antioxidant, antibacterial and brine shrimp lethality bioassay of Amoora cucullata using bark and leaf extracts of petroleum ether, chloroform and methanol. Methods: Bark and leaf parts were separately soaked into petroleum ether, chloroform and methanol respectively. To determine the presence of antioxidants; DPPH free radical scavenging assay, total phenolics and flavonoids determination and reducing power assay were carried out. Disk diffusion and brine shrimp lethality bioassay were conducted for the preliminary screening of antibacterial activity and cytotoxicity. Results: IC50 values in DPPH free radical scavenging assay for the extracts of petroleum ether, chloroform and methanol were found to be 316.23, 1192.42 and 128.82 in bark and 1106.32, 1330 and 25.12 μg/mL in leaf extracts respectively. Total phenolic content determined by Folin-Ciocalteu method in bark were 0, 0 and 58.41 mg gallic acid/gm of dry plant material in petroleum ether, chloroform and methanol while 8.11, 0 and 0 mg in leaf respectively. Total flavonoid content in bark and leaf was found to be 663.60, 549.47 and 46.25 mg and 1.64, 0 and 14.09 mg QE/gm of dried plant material in petroleum ether, chloroform and methanol respectively. In reducing power assay,methanolic extracts exhibited good reducing capacity. In antibacterial activity, extracts showed significant inhibition against enteropathogenic bacteria. Besides in brine shrimp lethality bioassay LC50 value was found to be 10, 2.301 and 7.28 in bark and 1.308, 1.94 and 2.14 μg/mL in leaf extracts of petroleum ether, chloroform and methanol respectively. Conclusion: From the present research work it can be concluded that A. cucullata leaf possess moderate antioxidant properties; strong antibacterial activity that correlates with their folkloric uses and potential cytotoxic properties.Key words: Antioxidant, Antibacterial, Cytotoxicity, Amoora cucullata.

BackgroundOleanolic acid (NZ-15), 7 α, 28-olean diol (NZ-38) and Stigmasterol (NZ-14) were isolated from the ethanolic extracts of the roots of Leea macrophylla (Family: Leeaceae) by using chromatographic analysis. This is the first report of isolation of these compounds from this plant. Their structures were constructed by spectroscopic analysis and by comparing the data with the published one. Subsequently the ethanolic extract was fractionated with two organic solvents and all the fractions were studied to evaluate their in vitro antioxidant property.MethodsThe ethanolic extract was fractionated with two organic solvents and all the fractions were studied to evaluate their in vitro antioxidant property by DPPH free radical scavenging assay, superoxide anion radical scavenging assay, nitric oxide radical scavenging assay, and reducing power assay.ResultsIn the DPPH free radical scavenging assay and superoxide radical scavenging assay, the ethyl acetate soluble fraction of ethanolic extract revealed the highest free radical scavenging activity with IC50 value of 2.65 and 155.62 μg/ml, respectively as compared to standard ascorbic acid (IC50 value of 5.8 and 99.66 μg/ml). Ethyl acetate fraction also possessed highest reducing power activity with an EC50 value of 15.27 μg/ml compared to ascorbic acid (EC50 0.91 μg/ml). On the other hand, the carbon tetrachloride fraction exhibited most significant NO scavenging activity with IC50 value of 277.8 μg/ml that was even higher than that of standard ascorbic acid (IC50 value 356.04 μg/ml). In addition, the total phenolic contents of these extract and fractions were evaluated using Folin-Ciocalteu reagent and varied from 7.93 to 50.21 mg/g dry weight expressed as gallic acid equivalents (GAE).ConclusionsThis study showed that different extracts of roots of L. macrophylla possess potential DPPH, superoxide, and NO free radical scavenging activities. The antioxidant activities of the plant extracts might be due to the presence of oleanolic acid, oleanolic acid derivative 7 α, 28-olean diol and stigmasterol.

Phenolic compounds profile and antioxidant activity of Dorystoechas hastata L. Boiss et Heldr

The larvicidal activity of crude petroleum ether, ethyl acetate, and methanol extracts of the whole plants of Phryma leptostachya L. was assayed for its toxicity against the early fourth instar larvae of Culex pipiens pallens. The larval mortality was observed after 24 h of exposure. Among three solvent extracts from Phyrma leptostachya L., the petroleum ether extract exhibited the best larvicidal activity. The corresponding LC₅₀ values of petroleum ether, ethyl acetate, and methanol extracts were 3.23, 5.23, and 61.86 ppm against the early fourth instar larvae of Culex pipiens pallens. The petroleum ether extract was successively subjected to column chromatography and preparative high performance liquid chromatography, and yielded the three lignans, phrymarolin-I, haedoxane A, and haedoxane E, which were isolated and identified as new mosquito larvicidal compounds. Phrymarolin-I, haedoxane A, and haedoxane E showed high larvicidal activity, for which the lethal doses LC₅₀ were estimated at 1.21, 0.025, and 0.15 ppm against the early fourth instar larvae of Culex pipiens pallens, respectively. The structures were elucidated by analyses of IR, UV, MS, and NMR spectral data. This is the first report on the mosquito larvicidal activity of the three compounds, phrymarolin-I, haedoxane A, and haedoxane E from Phyrma leptostachya L.