Chloroform Methanol Research Articles

Different Chromatographic Methods for the Determination of Antidiabetic Drugs in the Presence of Drug Toxic Impurity

Two validated, simple, and precise chromatographic methods were described for the determination of vildagliptin (VIL) and metformin (MET) in the presence of metformin toxic impurity, melamine (MEL). Method 1 is thin-layer chromatography (TLC)—densitometric method, at which methanol—chloroform—formic acid (7:3:0.3, by volume) was used as the developing system, and separation was carried out on Merck TLC silica gel 60 F254 aluminum sheets. The developed plates were air-dried and scanned at 215 nm. Linearity was constructed in the range of 0.2–2.6, 0.4–4.5, and 0.05–1.4 µg per band of VIL, MET, and MEL, respectively. Method 2 is reversed-phase ultra-performance liquid chromatography (RP-UPLC), where the separation was performed on a C18 column using methanol-acetonitrile—0.01 m sodium dihydrogen phosphate solution containing 50% 0.01 m sodium lauryl sulfate, pH = 5 with H3PO4 as the mobile phase at a flow rate of 1 mL min−1 at 205 nm. The calibration curves showed good linear relationships in the concentration ranges of 1–50, 2–70, and 0.5–30 µg mL−1 of VIL, MET, and MEL, respectively. The developed methods were applied to Galvus Met® tablets, and no interference from excipients was observed. The methods were validated as per the International Conference on Harmonisation (ICH) guidelines, and they compared favorably with the reported one.

Potential role of N-acetyl glucosamine in Aspergillus fumigatus-assisted Chlorella pyrenoidosa harvesting

BackgroundAlgal harvesting is a major cost which increases biofuel production cost. Algal biofuels are widely studied as third-generation biofuel. However, they are yet not viable because of its high production cost which is majorly contributed by energy-intensive biomass harvesting techniques. Biological harvesting method like fungal-assisted harvesting of microalgae is highly efficient but poses a challenge due to its slow kinetics and poorly understood mechanism.ResultsIn this study, we investigate Aspergillus fumigatus–Chlorella pyrenoidosa attachment resulting in a harvesting efficiency of 90% within 4 h. To pinpoint the role of extracellular metabolite, several experiments were performed by eliminating the C. pyrenoidosa or A. fumigatus spent medium from the C. pyrenoidosa–A. fumigatus mixture. In the absence of A. fumigatus spent medium, the harvesting efficiency dropped to 20% compared to > 90% in the control, which was regained after addition of A. fumigatus spent medium. Different treatments of A. fumigatus spent medium showed drop in harvesting efficiency after periodate treatment (≤ 20%) and methanol–chloroform extraction (≤ 20%), indicating the role of sugar-like moiety. HR-LC–MS (high-resolution liquid chromatography–mass spectrometry) results confirmed the presence of N-acetyl-d-glucosamine (GlcNAc) and glucose in the spent medium. When GlcNAc was used as a replacement of A. fumigatus spent medium for harvesting studies, the harvesting process was significantly faster (p < 0.05) till 4 h compared to that with glucose. Further experiments indicated that metabolically active A. fumigatus produced GlcNAc from glucose. Concanavalin A staining and FTIR (Fourier transform infrared spectroscopy) analysis of A. fumigatus spent medium- as well as GlcNAc-incubated C. pyrenoidosa cells suggested the presence of GlcNAc on its cell surface indicated by dark red dots and GlcNAc-specific peaks, while no such characteristic dots or peaks were observed in normal C. pyrenoidosa cells. HR-TEM (High-resolution Transmission electron microscopy) showed the formation of serrated edges on the C. pyrenoidosa cell surface after treatment with A. fumigatus spent medium or GlcNAc, while Atomic force microscopy (AFM) showed an increase in roughness of the C. pyrenoidosa cells surface upon incubation with A. fumigatus spent medium.ConclusionsResults strongly suggest that GlcNAc present in A. fumigatus spent medium induces surface changes in C. pyrenoidosa cells that mediate the attachment to A. fumigatus hyphae. Thus, this study provides a better understanding of the A. fumigatus-assisted C. pyrenoidosa harvesting process.

Estrogenic phytochemical from Labisia pumila (Myrsinaceae) with selectivity towards estrogen receptor alpha and beta subtypes

Labisia pumila var. alata (Myrsinaceae) or “Kacip fatimah” is a famous Malay traditional herb used for the maintenance of women's health. The extracts of L.pumila displayed estrogenic activity in rats. Nonetheless, the estrogenic bioactives were not identified. The aim of the study is to identify estrogenic compounds contributing to the established estrogenic activity. Bioactivity-guided-isolation method guided the isolation of pure bioactives. The hexane extract was subjected to a series of silica gel flash and open column chromatography with increasing amount of ethyl acetate in hexane or methanol in chloroform. Each fraction or pure compounds were evaluated on it's estrogen receptor (ER) binding activity with the fluorescence polarization competitive ERα and ERβ binding assay kit. Cytotoxic assay using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method was used to establish the cytotoxic activity of the compounds. Four alkyl resorcinols and a dimeric 1,4-benzoquinone, namely belamcandol B (1), 5-pentadec-10′-(Z)-enyl resorcinol (2), 1,3-dihydroxy-5-pentadecylbenzene (3), 5-(heptadec-12′-(Z)-enyl) resorcinol (4) and demethylbelamcandaquinone B (5) were identified with selective binding affinities towards either ERα or ERβ exhibiting selectivity ratio from 0.15–11.9. Alkyl resorcinols (2)–(4) exhibited cytotoxic activity towards HL60 cells with IC50 values from 19.5–22.0 μM. Structural differences between compounds influence the binding affinities to ER subtypes. Further study is needed to establish the agonist or antagonist effect of these compounds on various tissues and to identify if these compounds exert cytotoxic activity through the ERs. When consuming L.pumila as a complementary medicine, careful consideration regarding it's estrogenic compound content should be given due consideration.

Biocontrol of the pine processionary moth Thaumetopoea pityocampa (Denis andSchiffermuller, 1775) with plant extracts

Biocontrol of the pine processionary moth Thaumetopoea pityocampa (Denis and Schiffermuller, 1775) with plant extracts Kherroubi, M. Mouhouche, F. Zerrouk Izzeddine, Z. Chahbar, M. Key words Thaumetopoea pityocampa, LD50, Populus nigra, Taxus baccata Reception date: 15/03/2019 | Acceptation date: 06/06/2019 | Publication date: 04/0/2019 © Copyright belongs to the authors, who license the journal Arxius de Miscel·lània Zoològica to publish the paper under a Creative Commons Attribution 4.0 License, which permits its distribution and reproduction in any medium, provided the original authors and source, the journal Arxius de Miscel·lània Zoològica, are cited. Download: PDF | Cite Share on: | Biocontrol of the pine processionary moth Thaumetopoea pityocampa (Denis and Schiffermuller, 1775) with plant extracts Kherroubi, M. Mouhouche, F. Zerrouk Izzeddine, Z. Chahbar, M. Abstract Biocontrol of the pine processionary moth Thaumetopoea pityocampa (Denis and Schiffermuller, 1775) with plant extracts The purpose of this study was to propose alternative solutions based on the use of natural, bio-insecticidal products to fight against the processionary moth, Thaumetopoea pityocampa (T. pityocampa), considered a defoliator because of the damage it causes to silvicultural. This insect also causes health problems in forests or wooded areas open to the public. To meet this objective, we evaluated the toxicity of six extracts from two plants: Taxus baccata and Populus nigra, collected from the regions of Blida, against first instar larvae of T. pityocampa. The results from these biological tests showed a significant difference between the six extracts. The LD50 (lethal dose) values were calculated by probit analysis using Finney’s table. The LD50 was also low for the two extracts Populus Methanol and Populus Methanol Chloroform (1.4 mg/ml and 3.3 mg/ml, respectively). However, the LD50 was relatively high for the rest of the extracts. In conclusion, the Bio-insecticides of Populus Methanol (PMP) and Populus Methanol Chloroform (PMC) showed to be active and could be integrated into the biological control of the egg parasitoids of T. pityocampa.

ANTIMICROBIAL ACTIVITY OF THUJA ORIENTALIS IN COMBINATION WITH CIPROFLOXACIN AGAINST PSEUDOMONAS AERUGINOSA

OBJECTIVES: To compare the combined antimicrobial effect of Thuja orientalis and Ciprofloxacin against Pseudomonas aeruginosa in vitro samples. METHODS: In-vitro antibacterial activity of plant-extracts was evaluated alone and in combination with ciprofloxacin against Pseudomonas aeruginosa using disc-diffusion susceptibility assay (Kirby Bauer method) and minimum inhibitory concentration (96 well broth microdilution method) following CLSI guidelines. Pseudomonas aeruginosa clinical strains were collected from Rehman Medical Institute (RMI) and Northwest General Hospital Peshawar, Pakistan and ATCC strains (no.9721) of this bacterium were collected from Agriculture University Peshawar. The organism was tested six times with crude extract and fractionation with different solvents such as n-hexane, chloroform, ethyl acetate and butanol at concentrations of 1, 4, 8, 12, 16, 20, 24, 30 and 36 mg/ml. The mean MIC and FICI (fractional inhibitory concentration index) was obtained to report the synergism. The data were analysed using SPSS version-21. RESULTS: In combination, methanolic crude extract, chloroform and butanol fraction showed synergistic effect at all tested concentrations against Pseudomonas aeruginosa (ATCC 9721 and clinical) except with 1, 4, 16 mg/ml concentrations. Ethyl acetate and aqueous fractions shows indifference and synergistic effect against Pseudomonas aeruginosa (ATCC 9721 and clinical) at different concentrations. The fractional inhibitory concentration index (FICI) ranged from 1.24 to 3.24 against Pseudomonas aeruginosa alone and in combination with ciprofloxacin. CONCLUSION: By disc diffusion method, this study shows synergistic effect against Pseudomonas aeruginosa in combination with ciprofloxacin. However, through minimum inhibitory concentration method, it shows antagonism and indifference but no synergistic effect against different fractions of plant.

Biodiesel and biogas production from Isochrysis galbana using dry and wet lipid extraction: A biorefinery approach

Wet lipid extraction combined with residual biomass anaerobic digestion are alternatives to reduce the overall energy consumption of biodiesel production from microalgae. Solvents with different polarities have been studied to assess dry and wet lipid extraction process from Isochrysis galbana microalga. Ethyl acetate (EA) and a chloroform:methanol (CM) mixture yielded the best lipid extraction results in the dry and wet route with suitable lipid compositions. Fatty acid methyl esters (FAMEs) conversion of dry and wet extracted lipids with these solvents was performed by using both homogeneous (H2SO4) and heterogeneous (resin CT 269) catalysts. FAME production from wet extracted lipids with the EA solvent using the CT-269 resin constitutes an advantageous process because it avoids the water elimination step, and the CT-269 is a heterogeneous commercial catalyst, readily to separate after reaction. Lipid-spent microalga was anaerobically digested, obtaining that waste biomass from the wet extraction with EA had the highest methane yield (310 mL CH4/g volatile solids (VS). Energy balance analysis for FAMEs production with EA solvent (wet route) and heterogeneous catalyst yielded an energy recovery of about 80% in terms of biodiesel and biogas. Therefore, this process constitutes a promising route under an energy-driven microalga biorefinery.

The effects of different extraction methods of lipids from Nannochloropsis oceanica on the contents of omega-3 fatty acids

Nannochloropsis species are promising producers of the omega-3 fatty acid, eicosapentaenoic acid (EPA) of high interest for the food, nutraceutical and pharmaceutical industries. These applications demand the use of safer and food grade solvents. In this study, different solvent systems, including chloroform/methanol (CM), dichloromethane/methanol (DM), dichloromethane/ethanol (DE) and ethanol, were investigated as means of producing EPA-enriched lipid extracts from Nannochloropsis oceanica spray-dried biomass. Emphasis was given to the application of food grade ethanol, and ultrasound treatment was investigated as a means to increase lipid recovery during ethanol extraction. Total lipid extracts recovered with the different solvent systems were assessed by gravimetry and their lipid recovery efficiency determined by a purification process. The fatty acid composition of the total lipid extracts, particularly the EPA content, was determined by GC–MS.Of the various solvent systems tested the conventional CM and DM gave the highest yields with similar extraction efficiency followed by DE. Extraction with ethanol provided a total lipid extract of 19.1% dry biomass. Nevertheless, the combination of ethanol extraction with ultrasound treatment increased lipid yields, with an ultrasound probe providing the best results. The highest EPA content was obtained with CM and DM solvent systems followed by ethanol extraction assisted by ultrasound probe which increased EPA content by 35% relative to extraction with only ethanol, and comparable to that obtained with CM. These results indicate, for the first time, ethanol extraction assisted by ultrasound probe as a promising low toxicity, food grade and environmentally friendly approach to obtain EPA-enriched lipid extracts from N. oceanica biomass.

Antioxidant Potential, Phytochemicals Composition, and Metal Contents of Datura alba

This study investigated the phytochemical characteristics and antioxidant activity in leaves, roots, stem, flower, and seed parts of Datura alba (D. alba). The study also assessed the heavy metal (Cr, Mn, Zn, and Cu) accumulation in each part of the plant. Among the phytochemicals, alkaloids were found only in leaves while tannins, flavonoids, and phenols were present in all parts of the plant. For antioxidant activity, free radical scavenging assay for 2,2-diphenyl-1-picrylhydrazyl (DPPH) was performed using ascorbic acid as the standard. Higher activity was shown by stem extract in methanol and leaf extract in n-hexane, ethyl acetate, and chloroform. Furthermore, all the target heavy metals were detected in all plant sections with the highest concentration of Zn in leaves and Cu in stem, root, flower, and seed. Due to stronger antioxidant potential and phytochemical composition, D. alba could prove as valuable prospect in pharmaceutical formulations by taking part in the antioxidant defense system against generation of free radicals.

Comprehensive Fractionation of Antioxidants and GC-MS and ESI-MS Fingerprints of Celastrus hindsii Leaves.

Background: In this study, column chromatography was applied to separate active fractions from the ethyl acetate extract of Celastrus hindsii, a medicinal plant widely used in Southern China, Northern Vietnam, Myanmar, and Malaysia. Methods: Fourteen fractions from different dilutions of chloroform and methanol were separated by column chromatography and examined for biological activities. Results: It was found that a dilution of 50–70% methanol in chloroform yielded the highest total phenolics, flavonoids, and antioxidant activities (1,1-dipheny1-2-picrylhydrazyl (DPPH), 2,2-azinobis (3-ehtylbenzothiazoline-6-sulfonic acid), diammonium salt (ABTS) radical scavenging activity, and β-carotene bleaching method measured by lipid peroxidation inhibition). In addition, by gas chromatography-mass spectrometry (GC-MS) and electrospray ionization-mass spectrometry (ESI-MS) analyses, fifteen principal compounds from bioactive fractions belonging to fatty acids, amides, flavonoids, sterols, terpenes, and phenols were identified. Of these compounds, α-amyrin, β-amyrin, hydrazine carboxamide, hexadecanoic acid, fucosterol, (3β)-D:C-friedours-7-en-3-ol, rutin, and 2-hydroxy-1-ethyl ester accounted for maximal quantities, whilst concentrations of other constituents were <5%. Conclusions: It is suggested that these identified compounds may greatly contribute to the antioxidant capacity of C. hindsii as well as its potential pharmaceutical properties.

Enhanced separation of maximum boiling azeotropic mixtures with extractive heterogeneous-azeotropic distillation

In the separation industry the extractive heterogeneous-azeotropic distillation (EHAD) is a new and powerful innovation, that is capable of making the separation of highly non-ideal mixtures feasible and economical. In the last years there has been much attention paid to the separation of the minimum boiling homogeneous azeotropes. Although maximum boiling azeotropes are fewer in numbers than the minimum boiling ones but their separation is more complicated but it could be solved with the EHAD, too. Since EHAD is not limited to the separation of minimum boiling azeotropes, the separation of the maximum boiling azeotropes is studied in this work. Our work is motivated by industrial problems because there are such maximum boiling azeotropes in the liquid wastes of the fine chemical industry. The separation of highly non-ideal Water–Acetone–Chloroform–Methanol and Water–Ethyl Acetate–Chloroform–Ethanol quaternary mixtures are investigated and optimized in professional flowsheet simulator environment. Total Annual Costs are also determined. The purity requirement is 99.5 m/m% for Chloroform and the bottom product should be as clear as possible in water so that less liquid organic waste has to be incinerated. It is also an important merit of the EHAD that the chemicals in the distillate can be usually reused supporting sustainability. Different solutions for the separations supplemented with heat integration are examined. On the basis of the computer simulations and the experimental verification it can be concluded, the first time on the literature, that the separation efficiency of EHAD is superior also for the separation of the maximum boiling azeotrope mixtures.

Phytochemical constituents and antidiabetic activity of edible marine red seaweed (Halymenia durvilae)

Seaweeds have bioactive compounds with enormous health prospective which interests the pharmaceutical industries. The isolated bioactive compounds of seaweeds have been utilized as drug and food in the world. Phytochemical constituents of seaweeds have an assortment of prospective biological activity, such as antidiabetes. In worldwide the appearance of type two diabetes mellitus (T2DM) as the greatest non-transmittable disease has motivated search for new antidiabetic approaches. The purpose of the research was to determine the phytochemical properties and antidiabetic effect using α-glukosidase on methanol extract, n-hexane, chloroform, and water fraction of marine red algae Halymenia durvilae. The result of this study showed the phyto-constituent of H.durvilae includes steroids, flavonoids and triterpenoids are present in all extracts. Saponins and hyquinones showed their presence only in methanol extract. Alkaloid and tannin were not present in methanol extract and its fractions. H.durvilae on the extract and its fractions had antidiabetic activity. Water fraction had the highest activity to inhibit α-glukosidase (IC50 4.34±0.32 mg mL) followed by chloroform, hexane and methanol extract. Therefore, it can be concluded that H.durvilae could be used as a dietary food source of bioactive compound especially natural antidiabetic compounds.

Phytochemical screening and antibacterial activity of Aspilia africana on some gastrointestinal tract pathogens

The main aim of this study was to determine the phytochemical constituents and antibacterial activities of the leaf extracts of A. africana. The powdered leaf of A. africana was extracted using 70% methanol and partitioned into n-hexane, chloroform, ethyl acetate and aqueous methanol fractions. The extract and fractions were phytochemically screened and agar well dilution technique was used to evaluate the antibacterial activity against some gastrointestinal tract pathogens (Escherichia coli, Staphylococcus aureus, Shigella dysentriae, Salmonella typhi, Salmonella paratyphi A, Salmonella paratyphi B and Salmonella paratyphi C). The phytochemical analysis of A. africana showed the presence of alkaloids, terpenoids, tannins, flavonoids, anthraquinones and saponins in the methanol extract but varied across the fractions. The methanol extract and fractions inhibited the growth of gastrointestinal tract pathogens but their effectiveness varied with the concentrations. The methanol crude extract showed the best antibacterial activity with zone of inhibition ranging from 15-18 mm followed by aqueous soluble methanol fraction (10-15 mm) and ethylacetaate fraction (9-13 mm) respectively. The n-hexane and chloroform fraction showed no activity. Generally, the diameter zone of inhibition increased with increased extract concentrations. The antibacterial activity of the various extracts of the leaf was comparable to the reference antibiotics, though the antibiotics showed better inhibitory property (p<0.05) in some cases on the test isolates than the leaf extracts. The study showed that the leaves of A. africana possessed inhibitory properties due to the detected phytochemicals, thus validates the traditional uses of the plant for the treatment of gastrointestinal tract infections.

Metabolite profiling and wound-healing activity of Boerhavia diffusa leaf extracts using in vitro and in vivo models

Boerhavia diffusa is a perennial herb belonging to the Nyctaginaceae family. This plant has been widely used in Indian traditional medicinal system to cure several human ailments. However, traditional use of this plant in the treatment and management of wounds has not been validated by any comprehensive scientific study. The present study was aimed to explore the in vitro and in vivo wound healing potential of methanol extract (ME) and chloroform extract (CE) from B. diffusa leaf and subsequent identification of the bioactive metabolites, which might be responsible for enhancement of wound healing property of the extracts.The study included in vitro cell viability and wound scratch assays as well as in vivo excision wound assays in rat models. Both ME and CE were analysed for their antioxidant properties and phenolics content. The gas chromatography-mass spectrometry analyses were performed for identification of bioactive metabolites present in the ME and CE. ME of B. diffusa leaf significantly enhanced viability and migration of human keratinocyte cells (HaCaT) as compared to the untreated and CE-treated groups. The topical application of ME of B. diffusa leaf in excision wound model significantly decreased the wound area by the 14th day (91%) as compared to control (22%) (p < 0.05). The GC-MS analysis revealed the presence of caffeic acid, ferulic acid and D-pinitol as the major bioactive metabolites in ME. These results suggest that ME of B. diffusa possesses significant wound healing potential, where D-pinitol and caffeic acid served as the lead constituent metabolites responsible for the healing.

Exploration of a heat-integrated pressure-swing distillation process with a varied-diameter column for binary azeotrope separation

The pressure-swing distillation (PSD) processes with varied-diameter columns (VDCs) for separating methanol–chloroform and n-heptane–isobutanol are studied. Furthermore, two heat-integrated PSD processes, partial integration and full heat integration, are discussed with ordinary and VDCs. The results show that whether it is heat integrated or non-heat integrated, the processes using VDC have an advantage in the economy. Based on the minimum total annual cost (TAC), the dynamic controllability without and with full heat integration for an azeotrope system methanol/chloroform is explored. The dynamic controllability without and with partial heat integration for the azeotrope system n-heptane/isobutanol is discussed. The results indicated that compared with the dynamic responses without heat integration process, the heat-integrated PSD with a VDC did not increase the control difficulty while maintaining its economy. More azeotropic systems should be studied to investigate their economics and control effects, which will benefit PSD design and industrial application.

Ruthenium hydrazone complexes with 1:1 and 1:2 metal–ligand stoichiometry: a comparison of biomolecular interactions and in vitro cytotoxicities

Two ruthenium(II) complexes [RuIICl(PPh3)2(L)] (1) and [RuII(L)2] (2) were synthesized by reacting [RuCl2(PPh3)3] and thiophene-2-carboxylic acid (1-pyridine-2-yl-ethylidene)-hydrazide (HL) in methanol–chloroform, and characterized by elemental analysis and spectral and XRD data. The ratio of ligand to metal is 1:1 in the former complex and 2:1 in the latter. Interaction of these complexes with CT-DNA was studied using absorption and emission spectral studies; these show that both the complexes interact with CT-DNA through intercalative modes of interaction. Their BSA-binding activity results indicated the operation of static quenching mechanism and stronger binding of tryptophan residues than tyrosine residues. In vitro cytotoxicity assays against HeLa and MCF-7 cell lines showed better activity of both the complexes compared to the standard drug cisplatin. Overall, the activity of complex 2 with two units of coordinated ligand showed better activity than the other one.

Toxicological studies and bioactivity-guided identification of antimicrobially active compounds from crude aqueous stem bark extract of Boswellia dalzielii.

Objective:The main objective of this study is to isolate, identify, and quantify the active antimicrobial compounds present in the crude aqueous stem bark extract of B. dalzielii using some common pathogenic microorganisms as well as toxicological profile.Material and Methods:Crude aqueous stem bark extract of Boswellia dalzielii (CASEB) was partitioned by preparative thin layer chromatography (PTLC) using chloroform–methanol–water, 8:2:1 (v/v). The resulting bands were extracted using chloroform–methanol (50:50). The extract of each band was evaluated for antimicrobial activity on Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Klebsiella pneumonia, Pseudomonas aeruginosa, Proteus mirabilis, Salmonella typhi, and Candida albicans by disc diffusion. Compounds in the most antimicrobially bioactive fraction (MAAF) were identified by high performance liquid chromatography (HPLC), Fourier transform infrared spectrophotometry (FT-IR), and gas chromatography-mass spectrometry (GC-MS). Toxicological profile of the CASEB was evaluated by studying its effect in albino Wister rats.Results:PTLC produced five bands/fractions of which the MAAF was identified as RF2-fraction being active against all the isolates except E. coli and K. pneumoniae. HPLC of the MAAF revealed seven components; FT-IR revealed 17 functional groups; GC-MS revealed five compounds of which 93.18% are Oleic acid (44.88%), Squalene (34.16%), and n-Hexadecanoic acid (14.14%). The acute toxicity showed LD50 > 3,000 mg/kg. Sub-chronic toxicity showed that higher doses of the CASEB caused significant changes in liver function indices and a fatty change with lymphocytic infiltration (sign of acute hepatitis) in the liver tissues, but none of these changes were observed in the kidneys.Conclusion:The antimicrobially active compounds in CASEB were Oleic acid, Squalene, and n-Hexadecanoic acid. These can be further purified and used as precursors of new antimicrobial agents for treating infections especially those due to fungi and Pseudomonas spp. that are known to resist wide array of antimicrobial agents. The LD50 of CASEB is >3,000 mg/kg in rats. However, long-term consumption of CASEB is associated with significant liver damage.

Ameliorative potential of a traditionally used plant Fraxinus micrantha against oxidative stress and paracetamol-induced hepatotoxicity

Background: Liver disorders are one of the serious health issues. The treatment of liver disorders and their associated complications must be done with care due to the adverse effects of present-day medications. Therefore, there is an urgent need to develop effective and non-toxic herbal drugs for hepatoprotection. Objective: The objective of the present study is to investigate the hepatoprotective potential of traditionally used plant Fraxinus micrantha against paracetamol (PCM)-induced hepatotoxicity in rats. Methods: Hepatotoxicity was induced by a standardized single oral dose of paracetamol (PCM) at 3 g/kg body weight. Standard drug (silymarin 50 mg/kg) and test drugs (chloroform and methanol extracts) were administered orally to rats for 7 days. All the animals were sacrificed on the 8th day, and blood was withdrawn by retro-orbital vein puncture, collected in fresh centrifugation tubes, and centrifuged at 10,000 rpm for 10 min to separate serum for various estimations. Livers were isolated immediately for various biochemical and histopathological studies. Results: Treatment with chloroform extract (200, 400, and 800 mg/kg) and methanol extract (200 and 400 mg/kg) ameliorated the elevated levels of hepatic markers, but a significant reduction in these levels was observed by treatment with methanol extract at 800 mg/kg. In addition, treatment with chloroform and methanol extracts resulted in the amelioration of oxidative stress along with the histopathological changes. High-performance liquid chromatographic analysis of methanol extract of F. micrantha revealed the presence of various polyphenols such as rutin, naringenin, kaempferol, physicion, quercetin, and gallic acid. Conclusion: The results of the present study revealed that the observed hepatoprotective effect of methanol extract of F. micrantha may be due to the presence of various polyphenols and also provides pharmacological evidence for the use of F. micrantha bark in folk medicine for the treatment of liver diseases.

Chemical Composition and Antimycoplasmic Activity of Eugenia mattosii Leaves, Stems and Isolated Compounds

The Eugenia genus is rich in bioactive substances with biological potential. Mollicutes are bacteria without cell walls, which are responsible for various human and animal diseases. The aim of this study was to evaluate the chemical composition and antimollicute activity of Eugenia mattosii. Leaves and stems were extracted with methanol, partitioned into fractions of different polarities, and submitted to column chromatography in order to isolate major compounds. Identification and quantification of isolated substances in extracts and fractions were performed by UPLC-PDA/ESI+ - QTOF. Methanolic extracts, ethyl acetate and chloroform fractions and isolated substances were screened for antimycoplasmic activity against five Mycoplasma strains. The methanolic extracts of leaves and stems showed moderate antimycoplasmic activity (MICs = 250-500 μg.mL-1). The fractions exhibited better effect, with MICs = 125-1000 μg.mL-1, especially the ethyl acetate fraction of the leaves, which presented MICs of 125-250 μg.mL-1 for all strains tested. Phytochemical analyses evidenced the presence of some phenolic compounds, including pinostrobin, cryptostrobin and catechin, the first showing promising antimycoplasmic activity. Quantification of the compounds demonstrated higher concentrations of pinostrobin and cryptostrobin in the chloroform fraction. In conclusion, E. mattosii presented antimycoplasmic activity related, at least in part, to the presence of pinostrobin.

Free Radical Scavenging and Anticollagenase Properties of Sargachromanol I from Myagropsis myagroides

The present study investigated the free radical scavenging activity and antiwrinkle properties of methanol extract, hexane, chloroform, ethyl acetate, and butanol partition fractions of Myagropsis myagroides. The methanol extract showed higher free radical scavenging (65.46 % at 0.5 mg/ml) and collagenase inhibitory activity (29.64 % at 1000 μg/ml) than the water extract (16.68 % at 0.5 mg/ml and < 5 % at 1000 μg/ml). The hexane fraction exhibited strong free radical scavenging activity (96.36 % at 0.5 mg/ml) and collagenase inhibitory activity (IC50 of 712.21 μg/ml). The hexane fraction was subjected to silica gel column chromatography, Sephadex LH-20 column chromatography, octadecyl silica Sepak cartridge, and preparative high performance liquid chromatography. Sargachromaol I was isolated, which showed strong free radical scavenging activity (95.82 % at 0.1 mg/ml) and collagenase inhibitory activity (IC50 value of 54.01 μg/ml). The potential value in preventing skin aging could be confirmed through effects of extract from Myagropsis myagroides on free radical scavenging activity and collagenase inhibitory properties.

Antimalarial activity of hydromethanolic extract and its solvent fractions of Vernonia amygdalina leaves in mice infected with Plasmodium berghei.

Background:Vernonia amygdalina Del. (Asteraceae) is reported to be traditionally used for the treatment of malaria. Based on folkloric repute of this plant in Ethiopian traditional medicine and crude extract-based ethnopharmacological studies conducted in few countries, this study was undertaken to evaluate the in vivo antimalarial activity of 80% methanol extract and its solvent fractions of the leaves of V. amygdalina in mice infected with Plasmodium berghei.Methods:A 4-day suppressive test was conducted on mice infected with P. berghei to find out antimalarial effect of chloroform, butanol and aqueous fractions obtained from the 80% methanol crude extract. In all the activity tests, mice were randomly assigned in five groups (three tests and two controls) of six animals in each and received respective treatments. Data were analyzed using one way analysis of variance followed by Tukey’s post hoc test for multiple comparisons.Results:Acute oral toxicity test showed that all solvent fractions of the leaves of V. amygdalina revealed neither mortality nor overt signs of toxicity up to 2000 mg/kg. This study indicated that the percentage parasitemia suppression of 80% methanol extract was 32.47% (±2.65), 35.40% (±3.14) and 37.67% (±2.50) at 200, 400 and 600 mg/kg, respectively. All doses of the 80% methanol extract of V. amygdalina prolonged survival time and prevented weight loss and packed cell volume reduction in infected mice. All doses of chloroform and butanol fractions significantly suppressed parasitemia (p < 0.05), increased survival time (p < 0.05) compared to negative control and exhibited a significant reduction in rectal temperature (p < 0.05). All solvent fractions significantly prevented weight loss (p < 0.05) at all tested doses. The 80% methanol extract and chloroform and butanol fractions significantly (p < 0.05) prevented further reduction in rectal temperature of P. berghei-infected mice at all doses.Conclusion:The results of this study indicated that 80% methanol extract and solvent fractions of the leaves of V. amygdalina demonstrated promising antimalarial activity. The study corroborated the folklore use of this plant for the treatment of malaria in ethnomedicine in Ethiopia.