Activity Of Chloroform Fraction Research Articles

Evaluation of antiplasmodial potential of Cissampelos pareira L.: Characterization and quantification of isoquinoline alkaloids responsible for the biological activity

Globally, based on traditional knowledge, different parts of Cissampelos pareira plant are being used individually or in combination in various forms to manage malaria. However, the scientific investigation for validating the most effective part of this plant against malaria parasite has not been done. Thus, the study aims to explore the antiplasmodial potential of different parts of the plant against malaria parasite by bio-assay guided isolation of compounds from different extracts and fractions. Antiplasmodial activities of the whole plant parent extract and its fractions along with decoctions (roots, stem, leaves and whole plant) and isolated molecules were evaluated against chloroquine-sensitive Pf3D7 and chloroquine-resistant PfINDO strains of Plasmodium falciparum. A UPLC-DAD based method was developed to quantify the marker compounds in all samples by utilizing isolated molecules as analytical standards. The chloroform fraction of the whole plant was found to be the most potent with IC50 (µg/mL) of 0.79 (Pf3D7) and 2.26 (PfINDO) followed by root decoction with IC50 (µg/mL) 10.22 (Pf3D7) and 7.7 (PfINDO). Further, phytochemical analysis of active chloroform fraction led to the isolation of two potent bisbenzylisoquinoline compounds, namely O,O-dimethylcurine (1) [IC50 (µM) 1.46 (Pf3D7) and 0.51 (PfINDO)], and curine (2) [IC50 (µM) 1.46 (Pf3D7) and 0.51 (PfINDO)]. Analysis of developed UPLC-DAD method showed the highest quantities of curine (2) (∼107 mg/g) and O,O-dimethylcurine (1) (∼15 mg/g) in chloroform fraction which might be responsible for its potent activity. This study showed that the root decoction was more effective than decoctions of each of the other parts of the plant and the whole plant hydroalcoholic extract.

In vitro bioactivities and in silico enzyme interactions of abietatrien-3β-ol by bio-guided isolation from Nepeta italica subsp. italica

Abietatrien-3β-ol (ATO) was isolated from the aerial part of the Nepeta italica subsp. italica methanol-chloroform extract and studied antioxidant, enzyme inhibition, and DNA protection activities. The plant extract was fractionated by silica gel column chromatography using four different solvents. After the active chloroform fraction was divided into six subfractions under the guidance of bioactivity, ATO was isolated from the fourth active subfraction. The molecular structure of ATO was determined by the NMR techniques and confirmed with literature data. In the antioxidant test, ATO showed excellent DPPH˙ (IC50-1.18 ± 0.11 µg/mL), ABTS˙+ (IC50-1.82 ± 0.00 µg/mL), metal chelating (IC50-2.90 ± 0.05 µg/mL), superoxide anion scavenging (IC50-11.59 ± 0.27 µg/mL), reducing power (A0.5-21.09 ± 1.42 µg/mL), H2O2 (A0.5-57.81 ± 4.54 µg/mL), and phosphomolybdenum reducing (A0.5-124.23 ± 0.69 µg/mL) activities when compared to standards. The DNA protection potential was also found to be strong in Form I (47.89%) and Form II (4.56%) formations. ATO had high inhibitions in AChE (IC50-1.25 ± 0.04 µg/mL), BChE (IC50-1.26 ± 0.03 µg/mL), lipase (IC50-7.58 ± 0.27 µg/mL), and tyrosinase (IC50-9.60 ± 0.03 µg/mL). In molecular docking studies, ATO had a high binding affinity with α-amylase (−8.80 kcal/mol), tyrosinase (−8.10 kcal/mol), and urease (−8.10 kcal/mol) enzymes. In addition, by detailing the interaction of ATO with key enzymes that give the best interactions through molecular docking studies, with molecular dynamics studies, and by performing ADMET and DFT calculations, it was tried to explain that it could be a suitable drug candidate. In light of these studies, ATO is thought to be an effective enzyme inhibitor and a protective molecule against oxidation with its active antioxidant properties. Communicated by Ramaswamy H. Sarma

Antihyperlipidemic Bioactivity Guided Isolation and Structural Elucidation of Isolated Phytoconstituents from Convolvulus pluricaulis Choisy

Background: Atherosclerosis is a crucial element in the origination of coronary heart disease (CHD). Lipid aggregation inside the coronary arteries walls lead to the occurrence of hyperlipidemia. Aim: The research was conducted for assessing the lipid lowering effect of ethanol extract and chloroform fraction from Convolvulus pluricaulis Choisy. aerial parts in Triton induced hyperlipidemic rat model. Methodology: The active fraction was obtained from antihyperlipidemic bioactivity. The phytocomponents were separated from the active fraction using flash chromatography technique resulting in the isolation of n-hexatriacontane; 9-octadecenoic acid- octyl ester; 12, 14-heptacosanedione; dodecyl-octadeca-9,12-dienoate; tetracosanyl 9-hexadecenoate; heptacosan-14-ol; stigmasta- 5, 22- dien-3 beta-ol and stigmast-5-en-3 beta-ol. The phytocomponent structures were established by spectral data analysis. Intraperitoneal (i.p.) injection of Triton WR- 1339 at a dose of 400 mg/kg body weight (b.w.) was given to the animals. The ethanol extract at a dose of 200 mg/kg b.w. and chloroform fraction at a dose of 400 mg/kg b.w. were administered orally in rats after 24 hours of the Triton administration. Results: The study resulted in dose dependent inhibition of triglycerides (p <0.01), total cholesterol (p <0.05), Low density lipoprotein level (p <0.05) while significant increased HDL (p <0.01) level. Alteration in lipid profile in Triton treated rats was restored and found more effective in the chloroform fraction probably due to the presence of phytocomponents. The study elucidated different phytocomponents isolated from the active chloroform fraction of C. pluricaulis. Conclusion: Treatment with extract and fraction manifested significant decrease in triglyceride level. HDL exerts an inhibitory action in the etiology of atherosclerosis. It is considered as a beneficial lipoprotein and its lower level leads to the greater risk of CHD. It may be concluded that the isolated components were responsible for the lipid lowering level of active fraction. GRAPHICAL ABSTRACT

Unveiling the therapeutic potential: Evaluation of anti-inflammatory and antineoplastic activity of Magnolia champaca Linn’s stem bark isolate through molecular docking insights

Magnolia champaca Linn. has traditionally been used for medicinal activity in Asia for treating various chronic diseases as well as a source of food, medicines, and other commodities. Due to the long-used history of this plant, the present study was designed to explore the in vitro, in vivo and in silico anti-inflammatory and antineoplastic properties of the methanolic extract and fractions and the pure compound isolated from the most active chloroform fraction (CHF) of the stem bark of the plant. The isolated compound from the most active CHF was characterized and identified as a glycoside, trans-syringin, through chromatographic and spectroscopic (1H-NMR and 13C-NMR) analyses. In the in vitro anti-inflammatory assay, CHF was most effective in inhibiting inflammation and hemolysis of RBCs by 73.91 ± 1.70% and 75.92 ± 0.14%, respectively, induced by heat and hypotonicity compared to standard acetylsalicylic acid. In the egg albumin denaturation assay, CME and CHF showed the highest inhibition by 56.25 ± 0.82% and 65.82 ± 3.52%, respectively, contrasted with acetylsalicylic acid by 80.14 ± 2.44%. In an in vivo anti-inflammatory assay, statistically significant (p < 0.05) decreases in the parameters of inflammation, such as paw edema, leukocyte migration and vascular permeability, were recorded in a dose-dependent manner in the treated groups. In the antineoplastic assay, 45.26 ± 2.24% and 68.31 ± 3.26% inhibition of tumor cell growth for pure compound were observed compared to 73.26 ± 3.41% for standard vincristine. Apoptotic morphologic alterations, such as membrane and nuclear condensation and fragmentation, were also found in EAC cells after treatment with the isolated bioactive pure compound. Such treatment also reversed the increased WBC count and decreased RBC count to normal values compared to the untreated EAC cell-bearing mice and the standard vincristine-treated mice. Subsequently, in silico molecular docking studies substantiated the current findings, and the isolated pure compound and standard vincristine exhibited −6.4 kcal/mol and −7.3 kcal/mol binding affinities with topoisomerase-II. Additionally, isolated pure compound and standard diclofenac showed −8.2 kcal/mol and −7.6 kcal/mol binding affinities with the COX-2 enzyme, respectively. The analysis of this research suggests that the isolated bioactive pure compound possesses moderate to potent anti-inflammatory and antineoplastic activity and justifies the traditional uses of the stem bark of M. champaca. However, further investigations are necessary to analyze its bioactivity, proper mechanism of action and clinical trials for the revelation of new drug formulations.

Chemical and Biological Investigations of the Leaf Extracts of Andrographis paniculata (Acanthaceae)

Introduction: Andrographis paniculata is a plant employed in traditional medicine among the natives of Owerri, Eastern Nigeria, in the management of several ailments. In this study the leaf was investigated for its chemical constituents and to validate its antimicrobial activity in ethnomedicine. Methods: The fresh leaf of the plant was steam-distilled to obtain the volatile oil which was subjected to FT-IR and GC-MS analysis. The powdered air-dried leaf was also extracted with methanol to obtain the crude extract which was subjected to phytochemical analysis. The crude methanolic extract was also partitioned into hexane, chloroform, ethylacetate and 50% methanol in chloroform to obtain the corresponding fractions. The crude extract and fractions were tested for activity against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans and Aspergillus niger. The biologically active chloroform fraction was further purified using column chromatography and the fractions subjected to GC-MS analysis. Results: The steam distillation gave an oil and the GC-MS analysis indicated the presence of butylated hydroxytoluene, hexadecanoic acid, methyl ester, dibutyl phthalate, 8,11-octadecadienoic acid, methyl ester, 11-octadecenoic acid, methyl ester and bis (2-ethylhexyl) phthalate. Phytochemical analysis of the crude extract showed the presence of tannins, alkaloids, sterols/steroids, flavonoids, volatile oils, terpenoids, saponins and phenols. Among the extractives, the chloroform-soluble fraction showed the highest activity against S. aureus and B. subtilis while none of the extracts was active against C. albicans and A. niger. It was also found that the hexane-soluble fraction was inactive against all the test organisms while the ethylacetate-soluble fraction showed the broadest spectrum of anti-bacterial activity. Column chromatographic separation of the biologically active chloroform-soluble fraction gave fractions which on GC-MS analysis and by comparison with the standard library computer MS data led to the identification of some chemical constituents, including aromatic compounds, sterols, alicyclics and long-chain hydrocarbons and carbonyl compounds such as carboxylic acids, esters and lactones. Conclusion: The antimicrobial activity exhibited by the leaf extracts support the traditional medicinal use of the leaf to manage infections. Also, the presence of the identified phyto-constituents may be responsible for the ethno-medicinal properties of the plant.

Suppressive, Curative, and Prophylactic Effects of Maesa lanceolata Forssk. against Rodent Malaria Parasite Plasmodium berghei.

The artemisinin partial resistance is believed to be spread to artemisinin-based combination therapy partner drugs. As a result, new antiplasmodial compounds are required to treat resistant malaria infections. In the invention of antimalarial substances, claimed medical plants are precious resources. So, the current study was designed to assess the antiplasmodial effects of Maesa lanceolata in mice. In this study, preliminary phytoconstituent and in vivo acute oral toxicity tests were done. Early infection, established infection, and residual infection tests were employed to determine the antimalarial effects of the test drugs. Three doses (200, 400, and 600 mg/kg) of the extracts were provided orally to the test mice. Analysis of variance (one-way) followed by post hoc Tukey's test was used to analyze the difference between and within groups. Terpenoids, tannins, saponins, flavonoids, and alkaloids were detected in the phytochemical constituent analysis. Both 80% methanolic crude extract and solvent fractions had no toxic result at the 2000 mg/kg dose. All test drug doses suppressed parasite levels in a significant manner at all tests. The activity of chloroform fraction (maximum percentage suppression, 81.28%) overwhelms the crude extract activity. The curative effects of 80% methanolic crude extract, with a maximum of 80.22% parasitemia suppression, were greater than its suppressive and prophylactic effects. The 400 mg/kg dose of chloroform fraction resulted in a maximum survival period (18 days) than other doses of tested materials. The results of this investigation provide support for the activity of M. lanceolata leaf extract against malaria.

Antiplasmodial diterpenoid alkaloid from Aconitum heterophyllum Wall. ex Royle: Isolation, characterization, and UHPLC-DAD based quantification

Ethnopharmacological relevanceAconitum heterophyllum Wall. ex Royle is a traditionally important medicinal plant having numerous therapeutic actions as documented in Ayurveda. This plant is traditionally known for combating worm infestation, fever, respiratory tract disease, vomiting, diarrhoea, diabetes, skin disorders, anaemia, and joint disorders. Further, it has been used alone and in combination with other plants to prepare various anti-malarial formulations. However, there is no report on the assessment of its anti-plasmodial activity, and the metabolite(s) responsible for this activity. Aim of the studyThe main aim of this study was to conduct phytochemical investigation of A. heterophyllum roots for the preparation of extract, fractions, and isolation of pure molecules to identify active fractions/molecules responsible for the anti-plasmodial activity, and development of UHPLC-DAD based analytical method which can be used for the quantification of marker compounds in the extracts and fractions. Materials and methodsHydroalcoholic extract (1:1 v/v) and fractions (n-hexane, chloroform, ethyl acetate, n-butanol, and water) were prepared from the dried powdered roots of A. heterophyllum. Fractions were further subjected to silica gel column chromatography to isolate pure specialized secondary metabolites from this plant. All extracts, fractions, and pure molecules were evaluated against the chloroquine resistant Pf INDO and chloroquine sensitive Pf3D7 strains in culture for calculating their IC50 values. UHPLC-DAD based analytical method was also developed for the first time for the quantification of marker compounds and quality assessment of this commercially important Himalayan medicinal plant. ResultsPhytochemical investigation of A. heterophyllum root led to the isolation of six specialized metabolites viz. 2-O-cinnamoyl hetisine (1), atisinium (2), 4-oxabicyclo [3.2.2] nona-1(7),5,8-triene (3), atisinium cinnamate (4), aconitic acid (5), and atisinium formate (6). Compound 1 is a new hetisine type diterpenoid alkaloid, compounds 4 and 6 are new counter ionic forms observed with atisinium ion, and compound 3 is being reported for the first time from this genus. Chloroform fraction was found to be the most active with IC50 (μg/mL) 1.01 (Pf INDO) and 1.32 (Pf3D7). The molecule 2-O-cinnamoyl hetisine (1), a new diterpenoid alkaloid isolated from chloroform fraction, showed promising antiplasmodial activities with IC50 (μM) 1.92 (Pf INDO) and 10.8 (Pf 3D7). The activity of chloroform fraction was further validated by the developed UHPLC-DAD based method as the quantity of 2-O-cinnamoyl hetisine (1) was higher in the chloroform fraction (≅200 mg/g) than in all other fractions (<7 mg/g). Atisinium (2) and 2-O-cinnamoyl hetisine (1) were found to be the main marker compounds of this plant based on quantity and antiplasmodial activity, respectively. ConclusionThis study provides the scientific rationale for the traditional use of this plant in treating malaria. Further, this study revealed that the anti-malarial potential of this plant might be due to the presence of diterpenoid alkaloids.

Isolation and anti-cancer evaluation of two anti-proliferative constituents from the chloroform fraction of leaves of Conyza Sumatrensis (Retz.) E. H. Walker, Asteraceae

The plant Conyza sumatrensis, is reputed for anticancer activities in most parts of Nigeria. Our previous study on the anticancer evaluation of the extract and fractions of the plant on breast (MCF-7) and lung (NCI-H460) cell lines, revealed a higher sensitivity of the Chloroform (CHCl3) -fraction against the cell lines. The present study involved the bioassay guided fractionation of the active chloroform (CHCl3) fraction of the leaves of Conyza sumatrensis which yielded two known compounds coded as C110 and C219 and were subjected to MS,1D and 2D NMR spectroscopy as well as anti-proliferative studies. The identities of the compounds were revealed via NMR and MS as Stigmast-5, 22-dien-3-O-β-D-glucopyranoside (1) and 2, 3-dihydroxylpropyl hexacosanoate (2) whose GI50 were 40.83 ± 0.1, 22.67 ± 1.33 % and 58.83 ± 11.2 and 34 ± 5.6 % against MCF-7 and NCI-H460 cell lines respectively.The anti-proliferative activities of the compounds showed a considerable cell growth-inhibitory activity against human breast (MCF-7) and lung (NCI-H460) cell lines. These compounds are isolated from the plant for the first time and could serve as lead compounds for further investigation as anti-proliferative agents which could explain the rationale for its use in ethno-medicinal practice.

Studies on Analgesic and Anti-Inflammatory Activities of Aerial Parts of Tephrosia Bracteolata GUILL. and PERR. (Fabaceae) in Rodents

Background: Tephrosia bracteolata is a widespread shrub belonging to the family (Fabaceae) and genus Tephrosia. It is traditionally used for treating rheumatic pains, dropsy and stomach ache.Objectives: In view of the ethnomedicinal claim and the continuous search for new medicinal agents, the phytochemical constituents, analgesic and anti-inflammatory activities of chloroform fraction (CF) of the methanol extract of Tephrosia bracteolata in mice and rats was evaluated.Methods: Preliminary phytochemical screening was conducted using standard method. Analgesic activity of CF (100, 200 and 400 mg/kg body weight orally) was investigated using acetic acid-induced writhing test and thermally induced pain model in mice. Additionally, anti-inflammatory activity was tested by carragenaan-induced paw edema in rats.Results: Phytochemical screening revealed the presence of alkaloids, triterpenes and flavonoids. The oral LD50 of CF was above 2000 mg/kg body weight. CF significantly (p&lt;0.05) and dose dependently reduced the number of writhes with percentage inhibition of 47.76 48.41 and 72.6 % at dose of 100, 200 and 400mg/kg respectively. CF also significantly (p&lt;0.05) and dose dependently increased the mean reaction time. At dose of 400 mg/kg, CF at 60 and 90 minutes exhibited greater activity when compared to the standard agent pentazocine. CF(200 and 400 mg/kg) at times 3, 4 and 5 hours significantly (p&lt;0.05) decreased the paw edema in rats when compare with the ibuprofen treated group.Conclusions: The chloroform fraction of the methanol crude extract of Tephrosia bracteolata possesses analgesic and anti-inflammatory activities.

Anti-sickling activities of two isolated compounds from the root of Combretum racemosum P. beauv. (Combretaceae)

Ethnopharmacological relevanceEvaluation of plants such as Combretum racemosum with claimed traditional use in the management of sickle cell anaemia in Nigeria and other parts of West Africa could serve as a useful research strategy in the search for potential anti-sickling drugs and templates.Aim of the study: This study aimed at evaluating the antisickling potential of C. racemosum by activity-guided purification and isolation of its active constituents. Materials and methodsCrude methanol extract of the root of C. racemosum and the fractions obtained by partitioning with chloroform, ethyl acetate, and aqueous were investigated for anti-sickling activity against sodium metabisulphite induced sickling of sickle cell haemoglobin (HbSS). Repeated chromatographic separations were conducted on the most active chloroform fraction to purify and isolate bioactive compounds for further tests for anti-sickling activity. The characterization of the isolated compounds was done by mass spectrometry (FD+MS) and nuclear magnetic resonance (1HNMR) spectroscopy. ResultsThe chloroform fraction (FA) (% sickled erythrocyte ranged from 3.0 to 34.1) exhibited better anti-sickling activity than aqueous (% sickled erythrocyte ranged from 38.9 to 51.5) as well as the crude methanol (% sickled erythrocyte ranged from 19.1 to 30.4). Hence, the phytochemical investigation was focused on the chloroform fraction, which led to the identification of two ellagic acid derivatives (3,3′,4′-tri-O-methyl ellagic acid (A) and 3,3′-di-O- methyl ellagic acid (B). The two isolated compounds possessed good, comparable anti-sickling activities with compound A exhibiting a slightly better in vitro activity. ConclusionThis paper reports for the first time anti-sickling principles from C. racemosum and therefore, provided some justification for the ethnomedicinal use of the plant in the management of sickle cell disease.

Antibacterial, Antifungal and Anti-tubercular Activities of Chloroform Fraction of the Leaf Extract of Irvingia Gabonensis (African Bush Mango)

Background: The prevalence of anti-drug resistance by disease causing microorganisms has necessitated the search for alternative sources of drugs for the treatment of the ailments caused by these microorganisms. This study examines the biological properties of extracts from the leaves of Irvingia gabonensis (bush mango). Objective: The objective of this study is to determine the anti-microbial activity of chloroform fraction of the leaf extract and compare it with that of clinical reference. Methods: Antimicrobial activity of the chloroform fraction of the leaf extract of Irvingia gabonensis was evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Shigella dysenteriae, Salmonella typhi, Klebsiella pneumonia, Salmonella paratyphi, Candida albicans and Trichophyton rubrum by using the agar well diffusion method and Mycobacterium tuberculosis using agar proportion method on Lowenstein–Jensen medium. Preliminary phytochemical screening of the chloroform leaf fraction was done using qualitative standard methods. Results: This showed the presence of saponins, flavonoids, tannins, coumarin, phenol and alkaloids. Organisms were susceptible to chloroform fraction at different concentrations. The lowest MIC value obtained was 0.625mg/mL for S. aureus and S. typhi. While, five out of seven mycobacterial strains that were used, were susceptible. Conclusion: The antimicrobial activity is a result of the phytochemicals present in leaf. Therefore, we conclude that Irvingia gabonensis leaves can be used in the development of new pharmaceuticals research activities such as drug production.

Inhibitory activity of Sargassum hystrix extract and its chloroform fractions on inhibiting the α-glucosidase activity

Seaweed is one of the marine biotas that has several benefits for health and cosmetics. The objective of this study was to isolate and test the activity of chloroform fraction of Sargassum hystrix seaweed extract in inhibiting the α-glucosidase and identification of the active fraction compounds. S. hystrix was extracted by maceration, then partitioned using chloroform and methanol. The chloroform fraction was further separated by column chromatography to obtain purer compounds. The extract of S. hystrix, chloroform fraction resulting from partitioning and chloroform fractions from column chromatography were tested for inhibitory activity against the α-glucosidase. The active chloroform fraction inhibits the α-glucosidase then identified its compounds using Gas Chromatography-Mass Spectrophotometry (GC-MS). The result showed that chloroform fraction (IC50 = 0.726 mg/mL) had a higher inhibitory activity of the α-glucosidase than acarbose (IC50 = 1.890 mg/mL). Column chromatography separation produced seven chloroform fractions. The highest inhibition activity was found in the second chloroform fraction (41.62%), third (51.39%), and fourth (30.49%). Based on GC-MS analysis, several compounds were thought to be able to inhibit the α-glucosidase. The identified compounds were 2-Hexadecene-1-ol, 3,7,11,15-tetramethyl-; Hexadecanoic acid, methyl ester; 9-Octadecenoic acid, methyl ester; Phytol; Stigmasta-5,24(28)-dien-3-ol, (3.beta,24Z)- and 6-Hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydro benzofuran-2 (4H )-one.

Antidiabetic activities of chloroform fraction of Anthocleista vogelii Planch root bark in rats with diet- and alloxan-induced obesity-diabetes

Ethnopharmacological relevanceAnthocleista vogelii Planch is a medicinal plant traditionally used in West Africa for the management and treatment of diabetes mellitus. Aim of the studyTo determine the antidiabetic activities of chloroform fraction (CF) of Anthocleista vogelii Planch root bark in rats with diet- and alloxan-induced obesity-diabetes. Materials and methodsInhibitory activities of CF against α-amylase and α-glucosidase activities were determined in vitro. Three weeks old rats were fed with high-fat diet for 9 weeks to induce obesity prior to further induction of diabetes using alloxan (150 mg/kg body weight, i.p.). Blood glucose levels and body weight were measured every 7 days throughout the experiment. Glucose tolerance was assessed in normal and CF-treated rats on day 21. Terminal blood samples were collected from sacrificed animals for the measurement of serum insulin levels. Pancreases were excised from treated and untreated animals for histopathological examination. ResultsLCMS/MS chromatographic profile of CF via positive and negative modes revealed 13 and 23 compounds respectively. Further analysis revealed quebrachitol (QCT), loganin, sweroside, oleoside 11-methyl ester and ferulic acid, which have been previously reported for their antidiabetic activities, as constituents of CF. CF inhibited activities of α-amylase (IC50 = 51.60 ± 0.92 µg/ml) and α-glucosidase (IC50 = 5.86 ± 0.97 µg/ml) in a dose-dependent manner. Treatment of animals with obesity-diabetes with 100 and 200 mg/kg CF significantly improved glucose tolerance (P < 0.001) and enhanced serum insulin levels (P < 0.05) compared to diabetic control rats. ConclusionsAntidiabetic activities of CF might be mediated via inhibition of α-amylase and α-glucosidase activities, elevation of serum insulin concentration, and enhancement of insulin and leptin sensitivity in obesity-diabetes rats. This study further substantiates the traditional use of A. vogelii in the management and treatment of diabetes in Africa and encourages further studies to investigate its mechanism of action.

Evaluation of the anti-proliferative and cytotoxic potentials of fractions of methanol leaf extract of Cnidoscolus aconitifolius

The partitioned aqueous and chloroform fractions obtained from the methanol extract of the leaf of Cnidoscolus aconitifolius were examined for anti-proliferative (1-30 mg/mL) and cytotoxic activities (20-400 μg/mL) using the seed radicle inhibition and tadpole mortality assays over a period of 24 and 96 h respectively. The vacuum liquid chromatographic sub-fractions of the active chloroform fraction weresimilarly evaluated at 1-10 mg/mL and 5-20 μg/mL concentrations respectively. At 100 μg/mL the chloroform fraction produced 86.67 % mortality which increases to 100 % at 400 μg/mL where the aqueous fraction gave 36.67 %. The potency was further increased as the VLC sub-fractions C4-C6, gave 33 % cytotoxicity at 5 μg/mL and which later increased to 100 % at 10 μg/mL. At the end of 96 h, the control seed had average length of 32.32 ± 2.73 mm compared to 1.92 ± 0.36 and 0.52 ± 0.36 mm observed in the seeds treated with 20 and 30 μg/mL of the chloroform fraction indicating 94.01 and 98.39 % inhibition where as there was total radicle growth inhibition (100 %) of seeds treated with 5 mg/mL of the active VLC sub-fractions (C1-3, C4-C6 and C7). The results showed a significant cytotoxic and growth inhibitory effects of the plant extract, particularly the chloroform fraction and its sub-fractions which further corroborate the folkloric use of the plant in treating cancer. However, further chromatographic analysis of the active VLC sub-fraction and their respective biological studies using cancer cell-lines are needed to support this.Keywords: Cnidoscolus aconitifolius, Sorghum bicolor, anti-proliferative, cytotoxicity, methanol extract, aqueous fraction, chloroform extract , vacuum liquid chromatography,

Characterization of Cnidoscolus quercifolius Pohl bark root extract and evaluation of cytotoxic effect on human tumor cell lines

Objective: To evaluate the chemical components of active extract from Cnidoscolus quercifolius root bark and its cytotoxic potential against several tumor strains. Methods: The high-performance liquid chromatography with diode-array detection and 1H and 13C nuclear magnetic resonance spectroscopy of the extract were used to distinguish the existence of possible functional groups in the root bark extract. The in vitro cytotoxic activity of methanol extract on human colon cancer cell lines was evaluated using OVCAR-8, SF-295, HCT-116, HL-60 strains and the samples were assessed by 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide method. Results: The analysis of nuclear magnetic spectra of the active chloroform fraction revealed the presence of absorptions bands correspondent to a mixture of favelines such as neofavelanone, deoxofaveline or methyl-faveline, which structures were confirmed by ultraviolet spectra upon high-performance liquid chromatography with diode-array detection analysis. The active fraction showed cytotoxic effects in the tested strains, HCT-116, SF-295, OVCAR-8 and HL-60 cells with IC50 of 72 hours ranging from 4.95 to 15.23 μg/mL. Conclusions: The results suggest that the substances present in faveleira (Cnidoscolus quercifolius) root bark extract have a cytotoxic potential against several tumor lines, showing a broader antitumour potential, and in addition no adverse to healthy cells. Therefore, the root bark extract of Cnidoscolus quercifolius has a possibility of use for anticarcinogenic therapies.

Fraksi Kloroform Ekstrak Buah Mentimun (Cucumis sativusL.) Sebagai Anti Bakteri Terhadap Staphylococcus epidermidis

Penggunaan antibiotik secara besar-besaran di masyarakat memicu terjadinya resistensi terhadap bakteri, Untuk itu perlu adanya penelitian tentang alternatif lain terutama obat herbal sebagai antibakteri. Satu di antara tanaman berkhasiat obat adalah mentimun yang banyak terdapat di Indonesia.Tujuan penelitian ini adalah untuk membuktikan daya antibakteri fraksi kloroform ekstrak buah mentimun terhadap Staphylococcus epidermidis. Ekstraksi buah mentimun dilakukan dengan cara maserasi dengan etanol 70 %, selanjutnya ekstrak difraksinasi dengan kloroform. Fraksi yang digunakan adalah konsentrasi 5, 10, 15, 20, dan 25 %. Kemudian dilakukan uji daya antibakteri dan penentuan Konsentrasi Hambat Minimum. Hasil penelitian uji daya antibakteri fraksi kloroform ekstrak buah mentimun terhadap bakteri Staphylococcus epidermidis menunjukan adanya zona hambat pada konsentrasi 5, 10, 15, 20,dan25% dengan diameter masing-masing 5.86, 6.5, 7.05, 7.43 dan 9.92 mm. Selain itu, konsentrasi hambat minimum fraksi kloroform ekstrak buah mentimun terhadap S. epidermidis terdapat pada konsentrasi 3 %. Dari hasil kultur ulang didapatkan pertumbuhan pada media Nutrient Agar yang berarti fraksi kloroform ekstrak buah mentimun pada konsentrasi 3% bersifat bakteriostatik.

A novel steroid and cytotoxic constituents from Dioscorea membranacea Pierre against hepatocellular carcinoma and cholangiocarcinoma cells

Ethnopharmacological relevanceThe rhizomes of Dioscorea membrancea Pierre have been used in Thai traditional medicine as an ingredient formula for liver cancer and cholangiocarcinoma treatment. Aim of the studyTo investigate the cytotoxic activity of ethanolic extract and constituents of D. membrancea to support its traditional use. Materials and methodsThe SRB assay was used to determine the cytotoxic activity against hepatocellular carcinoma (HepG2), cholangiocarcinoma (KKU-M156) cells and one normal human keratinocyte immortal cells (HaCaT) with its ethanolic extract and isolated compounds. Bioassay guided isolation was used for isolating cytotoxic compounds. ResultsThe ethanolic extract of D. membranacea rhizome showed weak cytotoxic against KKU-M156 and HepG2 (IC50 at 72h exposure=30.49±0.82 and 38.97±2.04µg/mL respectively). A new steroid [epipanthogenin B (1)], a known steroid [panthogenin B (2)], two napthofuranoxepins [dioscorealide A (3) and dioscorealide B (4)], phenanthraquinone [dioscoreanone (5)] and two phenanthrene [5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (6) and 2,5,6-trihydroxy-3,4-dimethoxy, 9, 10-dihydrophenanthrene (7)] were isolated from active chloroform fraction. Compound 4 showed the highest cytotoxicity against HepG2 (IC50 at 72h exposure=2.87±0.21µM) and KKU-M156 (IC50 at 72h exposure=1.67±0.10µM) and less toxicity against normal cell line (HaCaT) (IC50 at 72h exposure>100µM). Compound 5 showed selective cytotoxic activity against KKU-M156 (IC50 at 72h exposure=3.46±0.11µM). Compounds 6 and 7 showed weak cytotoxic activity against HepG2 (IC50 at 72h exposure=24.96±2.32 and 51.31±3.52µM). Compounds (1–3) showed no cytotoxic activity against HepG2 and KKU-M156 cell lines (IC50 at 72 h exposure>100µM). ConclusionSeven compounds were isolated from active chloroform fraction of the ethanolic extract of D. membranacea rhizomes. Only dioscorealide B (4) might be served as a good anticancer agent for liver cancer and cholangiocarcinoma cancer because it can kill cancer cell but not toxic on normal cell. This research support Thai traditional medicine use of D. membranacea for liver cancer and cholangiocarcinoma cancer treatment.

Aqueous extract from the Withania somnifera leaves as a potential anti-neuroinflammatory agent: a mechanistic study

BackgroundMicroglial-mediated neuroinflammation is a key factor underlying the pathogenesis of various neurodegenerative diseases and also an important target for the development of the neuroinflammation-targeted therapeutics. Conventionally, the nonsteroidal anti-inflammatory drugs (NSAIDs) are prescribed, but they are associated with long-term potential risks. Natural products are the cornerstone of modern therapeutics, and Ashwagandha is one such plant which is well known for its immunomodulatory properties in Ayurveda.MethodsThe current study was aimed to investigate the anti-neuroinflammatory potential of Ashwagandha (Withania somnifera) leaf water extract (ASH-WEX) and one of its active chloroform fraction (fraction IV (FIV)) using β-amyloid and lipopolysaccharide (LPS)-stimulated primary microglial cells and BV-2 microglial cell line. Iba-1 and α-tubulin immunocytochemistry was done to study the LPS- and β-amyloid-induced morphological changes in microglial cells. Inflammatory molecules (NFkB, AP1), oxidative stress proteins (HSP 70, mortalin), apoptotic markers (Bcl-xl, PARP), cell cycle regulatory proteins (PCNA, Cyclin D1), and MHC II expression were analyzed by Western blotting. Mitotracker and CellRox Staining, Sandwich ELISA, and Gelatin Zymography were done to investigate ROS, pro-inflammatory cytokines, and matrix metalloproteinase production, respectively. Ashwagandha effect on microglial proliferation, migration, and its apoptosis-inducing potential was studied by cell cycle analysis, migration assay, and Annexin-V FITC assay, respectively.ResultsASH-WEX and FIV pretreatment was seen to suppress the proliferation of activated microglia by causing cell cycle arrest at Go/G1 and G2/M phase along with decrease in cell cycle regulatory protein expression such as PCNA and Cyclin D1. Inhibition of microglial activation was revealed by their morphology and downregulated expression of microglial activation markers like MHC II and Iba-1. Both the extracts attenuated the TNF-α, IL-1β, IL-6, RNS, and ROS production via downregulating the expression of inflammatory proteins like NFkB and AP1. ASH-WEX and FIV also restricted the migration of activated microglia by downregulating metalloproteinase expression. Controlled proliferation rate was also accompanied by apoptosis of activated microglia. ASH-WEX and FIV were screened and found to possess Withaferin A and Withanone as active phytochemicals.ConclusionsThe current data suggests that ASH-WEX and FIV inhibit microglial activation and migration and may prove to be a potential therapeutic candidate for the suppression of neuroinflammation in the treatment of neurodegenerative diseases.

Assessment of Annona reticulata Linn. leaves fractions for invitro antioxidative effect and antimicrobial potential against standard human pathogenic strains

Since from long time the plant, Annona reticulata Linn. is known for its beneficial effects. Leaves of A. reticulata were screened for phytochemicals and in vitro antioxidant, antibacterial and antifungal activity. The shade dried leaves were extracted with methanol and aqueous methanolic extract was partitioned successively with n-butanol, chloroform and acetone solvents. Methanolic extract was subjected to antioxidant screening using DPPH free radical scavenging activity and H2O2 scavenging activity. Antibacterial and antifungal activity of extract and fractions were analyzed on eight different clinical bacterial and fungal strains using agar well diffusion method and broth dilution method (MIC and MMC determination). The antioxidant activity showed that the extracts exhibited scavenging effect in concentration-dependent manner. The extract showed potent inhibitory effect against Bacillus subtilis and Escherichia coli bacterial strains while in case of fungal strains the maximum effect was observed against Candida blanki. The maximum zone of inhibition of n-butanol, chloroform and acetone fractions was observed against B. subtilis, and E. coli respectively while all fractions exhibited potent inhibitory effect against C. blanki. MIC and MBC values were determined for active samples, methanol extract and chloroform fraction against Staphylococcus aureus, B. subtilis, E. coli and Pseudomonas aeruginosa which revealed lower MIC and MBC values. The fungal strains Candida albicans, Saccharomyces cerevisiae and C. blanki were used to calculate MIC and MFC values for methanol extract and acetone fraction which demonstrated lower MIC and MFC values. The results provided evidence that the plant is richly supplied with numerous phytoconstituents that might indeed be potential sources of natural antioxidant, antimicrobial agents and supplementary food.

Bioassay-guided isolation of Poliovirus-inhibiting constituents from Zephyranthes candida

Context: Plants of the Zephyranthes genus are globally used in folk medicine. In a previous study, Zephyranthes candida Linn. (Amaryllidaceae) was identified as having antiviral properties; this led to anti-poliovirus assay-guided isolation of compounds from crude methanol extract of the plant.Objective: Isolation of anti-poliovirus constituents from Z. candida.Material and methods: Active chloroform fraction from crude methanol extract of Z. candida (whole plant) was subjected to bioassay-guided fractionation; repeated column and preparative thin layer chromatography led to isolation of active compounds. Chemical structures were identified using spectroscopic techniques. Using serial two-fold dilution of maximum non-toxic concentration (MNTC) of each compound (0.0625–1 µg/mL for lycorine and 0.625–10 µg/mL for trisphaeridine and 7-hydroxy-3′,4′-methylenedioxyflavan), the ability of extracts to inhibit viral-induced cell death in tissue culture was evaluated 72 h post-infection by the colorimetric method using MTT (3-[4,5-dimethylthiazol–2-yl]-2,5-diphenyltetrazolium bromide) dye. Regression analysis was used to determine 50% inhibitory concentration (IC50) and 50% cytotoxicity concentration (CC50), from which selective index (SI) was calculated.Results: From the chloroform fraction, three compounds were isolated and identified, namely lycorine (1), trisphaeridine (2), and 7-hydroxy-3′,4′-methylenedioxyflavan (3) as the anti-polioviral components. Lycorine was the most active, with an IC50 value of 0.058 µg/mL followed by trisphaeridine (2) with an IC50 of 0.1427 µg/mL, and 7-hydroxy-3′,4′-methylenedioxyflavan (3) with an IC50 of 0.2384 µg/mL.Discussions and conclusions: The antipoliovirus activity of trisphaeridine (2) and 7-hydroxy-3′,4′-methylenedioxyflavan (3) is established in this report; these compounds are of moderate toxicity and have very good SI. They could be a potential template for the development of a new antiviral agent.