Ethyl Acetate-soluble Portion Research Articles

Protective effect of the ethyl acetate fraction of Qinghai-Tibet Plateau medicinal plant Chrysosplenium axillare Maxim. against ANIT-induced cholestatic liver injury in mice

BackgroundChrysosplenium axillare Maxim. has been used as a traditional Tibetan herb medicine for hepatobiliary disorder in Qinghai-Tibet Plateau for more than one thousand years. Hypothesis/purposewe hypothesized that the ethyl acetate-soluble portion extracted from C. axillare (ECA) could exert hepatoprotection effect against α-naphthylisothiocyanate (ANIT)-induced hepatotoxity and acute intrahepatic cholestasis in mice and aimed to investigate its potential mechanism. Study designmale KM mice with acute intrahepatic cholestasis induced by ANIT were orally administered with the ECA (100 and 50 mg/kg B.W.). Mice receiving vehicle (0.5% CMC-Na) served as control. Methods48 h after ANIT administration, mice were sacrificed. Blood was collected for serum enzymes and components analysis using commercial kits. Livers were removed for histopathological examination and protein preparation. The protein expression levels of farnesoid X receptor (FXR), bile-salt export pump (BSEP), multidrug resistance associated protein2 (MRP2), Na+-taurocholate co-transporting polypeptide (NTCP), small heterodimer partner (SHP), and cholesterol 7α-hydroxylase (CYP7A1) were evaluated by a western blot analysis. A phytochemical analysis was conducted to characterize the chemical constituents of ECA. Resultsoral administration of ECA at both doses (P < 0.05 or P < 0.01) can prevent ANIT-induced increases in levels of serum enzymes and components, including AST, ALT, ALP, DBIL, TBA, and TBIL and improve the liver pathologic change induced by ANIT exposure. Western blot analysis showed that the expression levels of FXR, BSEP, MRP2, NTCP, and SHP in ECA (100 and 50 mg/kg) groups were up-regulated versus model group (P < 0.05 or P < 0.01), and those of CYP7A1 in both ECA treated groups were down-regulated versus model group (P < 0.01). One new highly O-methylated flavonoids, chrysosplenoside I (1), together with fourteen analogues (2–15), was isolated from ECA and structurally identified by spectra analysis. Among them, the two abundant ingredients, chrysosplenosides A and I (11 and 1), were quantitatively determined to have a total content account for 32.2% of ECA. ConclusionECA protected against mice liver damage with acute intrahepatic cholestasis induced by ANIT via regulation of the expression of hepatic proteins involved in bile acids synthesis, transportation and re-absorption. These results confirmed the hepatoprotective efficacy of traditional Tibet herb medicine C. axillare and provided the first insight into its related mechanism as well as the potential active ingredients.

Chemical Studies and Antibacterial Activity of the Root of Rumex abyssinicus

One of the Ethiopian endemic plants used traditionally as medicine for different diseases is Rumex abyssinicus. It is widely known as ‘Meqmeqo’ in Amharic. The aim of this study was to isolate and characterize compounds from the root of R. abyssinicus and to test their bioactivities of solvent fractions of the extract. The crude extract was extracted from dried powdered root of the plant with methanol. Then it was fractionated with n‑hexane and ethyl acetate with increasing polarity. The yellow powder obtained from ethyl acetate fractionation was applied to column chromatographic separation that led to the isolation of two compounds, diisobutyl phthalate and emodine. Finally, antibacterial evaluation of the ethyl acetate and methanol soluble portions were carried out against four pathogenic bacteria. Both methanol and ethyl acetate soluble portions inhibited all selected bacterial in better at high concentrations.

Anti-epileptic potentials of the partitioned fractions of chamaecrista mimosoides

This research work aimed to establish scientific basis for the use of Chamaecrista mimosoides, in traditional medicine as anti-epileptic medication. The whole plant part of Chamaecrista mimosoides was extracted with ethanol and screened for phytochemicals. Acute toxicity study was carried out using Lorke’s method and the antiepileptic activity was evaluated using maximal electroshock induced seizure test in day-old chicks, pentylenetetrazole (PTZ) and strychnine using mice. The phytochemical study revealed the presence of saponins, cardiac glycosides, tannins, flavonoids, terpenoids and cardenolides. Both the chloroform, ethylacetate and n-butanol portions at 100, 250, and 500mg/kg body weight did not protect the chicks against tonic hind limb extension (THLE) in maximal electro-shock test (MEST). The chloroform and n-butanol portions at doses of 250 and 500 mg/kg body weight protected 40% and 60% of mice against clonic spasm induced by pentylenetetrazole, while ethyl-acetate soluble portion did not protect the mice against clonic spasm induced by pentylenetetrazole at all doses used when compared to Valproic acid (200 mg/kg) protected all the mice (100%) against clonic spasm induced by pentylenetetrazole. The chloroform soluble portion at the doses of 100, 250 and 500 mg/kg body weight protected 40%, 100%, 100% against death induced by strychnine, while ethylacetate and n-butanol portions did not protect the rats against death induced by strychnine but prolonged the onset of convulsion. In all the tests, phenobarbitone (20 mg/kg) was used as positive control and protected 80% of mice against convulsion induced by strychnine. The antiepileptic investigation suggests that the chloroform portion of Chamaecrista mimosoides has a promising antiepileptic activity. Â

Chemical constituents from the leaves of Zanthoxylum nitidum (Roxb.) DC

Twelve compounds were isolated from the ethyl acetate soluble portion of the 70% hydroalcoholic extract from the leaves of Zanthoxylum nitidum (Roxb.) DC. All of the compounds were determined on the basis of UV, IR, NMR and MS and comparison with literature. Among them, compounds 4, 7, 10 and 12 were first acquired from Rutaceae, compound 2 was isolated from the genus of Zanthoxylum for the first time and compounds 1, 3, 6, 9, 11 were isolated from Z. nitidum for the first time. Our present study exhibited the taxonomic relationships between Z. nitidum and other species in the same genus; those between Rutaceae and other families. Furthermore, we have summarized the chemotaxonomic significance of these compounds in Rutaceae and Zanthoxylum.

New triterpenoids from Leonurus japonicus (Lamiaceae)

Three new triterpenoids (1, 4, and 5), together with 17 known analogues, were isolated from the ethyl acetate soluble portion of the EtOH extract of Leonurus japonicus Houtt. Their structures were determined by spectroscopic analysis. All known triterpenoids were isolated from the genus Leonurus for the first time. Among them, triterpenoids 2–3, 9–12, and 16–20 were first isolated from the family Lamiaceae. Furthermore, the cycloartane, taraxastane, ursane, lupane, euphane, and dammarane types are reported here for the first time from the genus Leonurus.

New abietane diterpenoid glucosides from underground parts of Isodon taliensis

Two new abietane diterpenoid glucosides, talienosides A and B (1 and 2), along with four known compounds (3 – 6), were isolated from the ethyl acetate soluble portion of 70% acetone extract of the underground parts of Isodon taliensis. Their structures were elucidated based on the analyses of extensive spectroscopic data and physicochemical properties. The two new compounds were tested for their bioactivities against malaria and diabetes by β-hematin formation inhibition and α-glycosidase inhibition experiments, respectively. However, only 2 showed weak β-hematin formation inhibitory activity.

Anti-inflammatory and bronchodilatory constituents of leaf extracts of Anacardium occidentale L. in animal models

ObjectiveAnacardium occidentale L. leaf is useful in the treatment of inflammation and asthma, but the bioactive constituents responsible for these activities have not been characterized. Therefore, this study was aimed at identifying the bioactive constituent(s) of A. occidentale ethanolic leaf extract (AOEL) and its solvent-soluble portions, and evaluating their effects on histamine-induced paw edema and bronchoconstriction. MethodsThe bronchodilatory effect was determined by measuring the percentage protection provided by plant extracts in the histamine-induced bronchoconstriction model in guinea pigs. The anti-inflammatory effect of the extracts on histamine-induced paw edema in rats was determined by measuring the increase in paw diameter, after which the percent edema inhibition was calculated. The extracts were analyzed using gas chromatography-mass spectrometry to identify the bioactive constituents. Column chromatography and Fourier transform infrared spectroscopy were used respectively to isolate and characterize the constituents. The bronchodilatory and anti-inflammatory activities of the isolated bioactive constituent were evaluated. ResultsHistamine induced bronchoconstriction in the guinea pigs and edema in the rat paw. AOEL, hexane-soluble portion of AOEL, ethyl acetate-soluble portion of AOEL, and chloroform-soluble portion of AOEL significantly increased bronchodilatory and anti-inflammatory activities (P < 0.05). Oleamide (9-octadecenamide) was identified as the most abundant compound in the extracts and was isolated. Oleamide significantly increased bronchodilatory and anti-inflammatory activities by 32.97% and 98.41%, respectively (P < 0.05). ConclusionThese results indicate that oleamide is one of the bioactive constituents responsible for the bronchodilatory and anti-inflammatory activity of A. occidentale leaf, and can therefore be employed in the management of bronchoconstriction and inflammation.

Antiplasmodial Compounds from Leaves of Dodonaea angustifolia.

Background: Dodonaea angustifolia is used in Ethiopian traditional medicine to treat malaria. The objective of this work was to conduct bioassay guided fractionation of the leaves of D. angustifolia using Plasmodium berghei infected mice.Method: The antiplasmodial activity of the extracts and pure compounds was evaluated using the standard Peter’s four-day suppressive method. The structures of isolated compounds were elucidated using chemi-cal and spectroscopic methods.Results: In this study, the ethyl acetate soluble portion of the 80% aqueous MeOH extract of the leaves significantly suppressed parasitaemia in Plasmodium berghei infected mice (80.28% at 150 mg/kg). Three active compounds which exhibited significant percent suppression of parasitaemia by 81% at 40 mg/kg, 80% at 50 mg/kg and 70% at 40 mg/kg, respectively were identified. These are the flavanone pinocembrin (1), the flavanol santin (2) and the clerodane diterpene 2-hydroxy-15,16-epoxyceloda-3,13(16),14-trien-18-oic acid (3). Under similar conditions, chloroquine suppressed parasitaemia by 100% at 25 mg/kg. Chemical study of the ethanol extract of the leaves yielded 5,7,4'-trihydroxy-3,6-dimethoxyflavone (4), ent-16-hydroxy-labdan-3α,8β-dihydroxy,13(14)-en-15,16-olide (5) and 5,6,7-trihydroxy-3,4'-dimethoxyflavone (6). Compound 6 has not been reported before as a natural product.Conclusion: From the leaves of D. angustifolia, three compounds with significant antiplasmodial activi-ties were isolated and characterized, with pinocembrin as the most active compound.

Hydrate Antiagglomeration Performance for the Active Components Extracted from a Terrestrial Plant Fruit

A terrestrial plant fruit extract was adopted to prevent hydrate accumulation. Four groups with different polarities, water-soluble, n-butanol-soluble, ethyl acetate-soluble, and petroleum ether-soluble portions, were obtained by prefractionation of the plant-extract hydrate antiagglomerant (AA). The hydrate antiagglomeration effect was tested in a high pressure transparent sapphire cell, and the active components were found mainly in the ethyl acetate-soluble portion that accounts for approximately 4.0% of the whole plant extract. Through further separation and purification, the five kinds of components obtained proved experimentally to have the effect of preventing hydrate agglomeration. Their molecules and structural formulas were inferred by the high resolution mass spectrum and nuclear magnetic resonance analysis. Compared with the spectrum library, the components were determined to be eriodictyol, apigenin, naringenin, luteolin, and 5-(4-hydroxy-6,7-dimethoxy-3-methylchroman-2-yl) benzene-1,2,3-trio...

울릉 마가목의 클로로겐산 이성체의 최종당화산물의 생성 저해 및 라디칼 소거 활성

Advanced glycation end product (AGE) formation and reactive oxygen species are potential therapeutic targets for the prevention of diabetic nephropathy and other pathogenic complications. Activity-guided isolation of an ethylacetate-soluble portion of 80% methanolic extract from fruits of Sorbus commixta of the Ulleung Island origin using AGE formation inhibition assay led to the isolation and identification of three caffeoylquinic acid derivatives of a previously known structure, 3-O-caffeoylquinic acid (neochlorogenic acid; 1), 4-O-caffeoylquinic acid (cryptochlorogenic acid; 2), and 5-O-caffeoylquinic acid (chlorogenic acid; 3). The structures of these compounds were confirmed by interpretation of nuclear magnetic resonance and mass spectrometry data. Among the isolates, the major metabolite, neochlorogenic acid (1) showed the most potent inhibitory effect against AGE formation with an IC50 value of 167.5±3.5 μM. Furthermore, all isolated chlorogenic acid isomers were evaluated for their radical scavenging activity against peroxynitrite, and structurally related isomers 1, 2, and 3 exhibited potent inhibitory effects in this radical scavenging assay. This result suggests that the monocaffeoyl quinic acid derivatives isolated from S. commixta might be beneficial for the regulation of diabetic complications and related diseases.

Spirostanols from the roots and rhizomes of Trillium tschonoskii

Two novel spirostanols, (23S,24R,25S)-18-norspirost-1,4,13-triene-21,23,24-triol-3,15-dione (1) and (23S,24S,25S)-spirost-5-ene-1β,3β,21,23,24-pentaol (2), a new natural product (3), and two known analogues (4 and 5) were isolated from the ethyl acetate-soluble portion of the ethanolic extract of Trillium tschonoskii Maxim. Their structures were elucidated by extensive spectroscopic analyses, and their cytotoxic activities on four kinds of human tumor cells were studied in vitro. Compound 4 showed significant cytotoxic activity against MCF-7 and A549 with IC50 values of 6.16±2.21 and 28.5±11.5μM, respectively, while 5 exhibited selective cytotoxicity against A549 with an IC50 value of 13.0±4.51μM.

Chemical Constituents of the Culture Broth of Phellinus linteus and Their Antioxidant Activity

The medicinal fungus Phellinus linteus, in the family Hymenochaetaceae, has been used as a traditional medicine for the treatment of various diseases. In this study, the chemical constituents of the culture broth of P. linteus were investigated. P. linteus was cultured in potato dextrose broth medium, and the culture broth was extracted with ethyl acetate. The ethyl acetatesoluble portion was concentrated and subjected to ODS column chromatography, followed by Sephadex LH-20 column chromatography. Six compounds (1~6) were purified by preparative reversed-phase high-performance liquid chromatography. Spectroscopic methods identified their structures as caffeic acid (1), inotilone (2), 4-(3,4-dihydroxyphenyl)-3-buten-2-one (3), phellilane H (4), (2E,4E)-(+)-4'-hydroxy-γ-ionylideneacetic acid (5), and (2E,4E)-γ-ionylideneacetic acid (6). Compounds 1, 2, and 3 exhibited potent dose-dependent antioxidant activity.

Penchinones A–D, two pairs of cis-trans isomers with rearranged neolignane carbon skeletons from Penthorum chinense

Two pairs of cis-trans isomers with new rearranged neolignane carbon skeletons from Penthorum chinense.

Penthorin A and B, two unusual 2,4′-epoxy-8,5′-neolignans from Penthorum chinese

Two new 2,4′-epoxy-8,5′-neolignans (1 and 2), together with five known 7,9′;7′,9-diepoxylignans (3−7), were isolated from an ethyl acetate soluble portion of a hepatoprotective water decoction of Penthorum chinese. Their structures and absolute configurations were elucidated by extensive spectroscopic analyses and electronic circular dichroism (ECD) calculations. This is the second report of 2,4′-epoxy-8,5′-neolignans from plants. Compounds 2, 6, and 7 showed in vitro protective activities against acetaminophen-induced hepatocyte injury at 5μM.

지모 추출물의 항산화 및 pancreatic lipase 저해 활성 평가

In this study, the antioxidant and pancreatic lipase inhibitory activities of aqueous methanolic (70% methanol) extract from the roots of Anemarrhena asphodeloides were investigated. The extracts of four solvent fractions (the n-hexane layer, EtOAc layer, n-BuOH layer, and H2O layer) of the 70% methanol extract were also investigated. Furthermore, the total phenolic content was quantified using a spectrophotometric method. All the tested samples showed dose-dependent radical scavenging and pancreatic lipase inhibitory activities. In particular, the pancreatic lipase inhibitory activity of the ethyl acetate soluble portion (the EtOAc layer) from the rhizomes of the A. asphodeloides was higher than that of the other solvent-soluble portions. The antioxidant property of the extracts was evaluated using radical scavenging assays with DPPH and ABTS+ radicals. 1000 mg/ml of the n-BuOH layer extract showed 91.2% DPPH radical scavenging activity. The EtOAc layer extract and the n-BuOH layer extract showed IC50 = 20.5±1.7 mg/ml and IC50 = 50.5±0.7 mg/ml ABTS+ radical scavenging activities, respectively. The anti-obesity efficacy of the A. asphodeloides extract was tested via porcine pancreatic lipase assay. A pancreatic lipase inhibitory activity (IC50) of 31.3±0.1 mg/ml was obtained from the EtOAc layer extract. These results suggest that A. asphodeloides can be considered a new potential source of natural antioxidant and anti-obesity agents.

Neuraminidase Inhibitors from the Fermentation Broth of Phellinus linteus

During a search for neuraminidase inhibitors derived from medicinal fungi, we found that the fermentation broth of Phellinus linteus exhibited potent neuraminidase inhibitory activity. Through bioassay-guided fractionation, two active compounds were purified from the ethyl acetate-soluble portion of the fermentation broth of P. linteus. These structures were identified as inotilone (1) and 4-(3,4-dihydroxyphenyl)-3-buten-2-one (2) by spectroscopic methods. Compounds 1 and 2 inhibited H1N1 neuraminidase activity with IC50 values of 29.1 and 125.6 μM, respectively, in a dose-dependent manner. They also exhibited an antiviral effect in a viral cytopathic effect reduction assay using MDCK cells. These results suggest that compounds 1 and 2 from the culture broth of P. linteus would be good candidates for the prevention and therapeutic strategies towards viral infections.

Antiplasmodial anthraquinones and hemisynthetic derivatives from the leaves of Tectona grandis (Verbenaceae)

Chemical investigation of the methanol extract of the leaves of Tectona grandis led to the isolation of one new anthraquinone derivative, grandiquinone A (3-acetoxy-8-hydroxy-2-methylanthraquinone) (1), along with nine known compounds: 5,8-dihydroxy-2-methylanthraquinone (2), hydroxysesamone (3), 3-hydroxy-2-methylanthraquinone (4), quinizarine (5), betulinic acid (6), ursolic acid (7), tectograndone (8), corosolic acid (9) and sitosterol 3-O-β-d-glucopyranoside (10). Compounds 2 and 3 were isolated for the first time from the leaves of this plant, while 5 has never been reported from the genus Tectona. Hydroxysesamone (3) and tectograndone (8) were subjected to cyclisation and acetylation reactions to afford two hemisynthetic derivatives, 6,9-dihydroxy-2,2-(dimethyldihydropyrano)-3,4-dihydro-2H-benzo[g]chromene-5,10-dione (11) and acetyltectograndone (12) respectively, which are reported here for the first time. The ethyl acetate-soluble portion, some of the isolated compounds and hemisynthetic derivatives were evaluated for their antiplasmodial activity against the multidrug-resistant Dd2 strain of Plasmodium falciparum. Compound 3 showed a prominent activity, while 2, 8, 9, 11 and 12 showed significant in vitro anti-malarial activity. Compound 1 was weakly active in this test. The structures of the compounds were elucidated by spectroscopic methods and comparison of the data with the literature.

Isolation of Gallic Acid and Estimation of Total Phenolic Content in Some Medicinal Plants and Their Antioxidant Activity

Gallic acid was isolated from ethylacetate soluble portion of the methanol extract of fruit pulp of Terminalia chebula and characterized by comparing the melting point, Rf values, UV and IR spectra with authentic gallic acid. Quantitative determination of phenolics from different extracts of medicinal herbs such as Adhatoda vasica Nees, Bergenia ciliata (Haw) Sternb, Phyllanthus emblica Linnaeus, Terminalia bellirica (Gaerth) Roxb, Terminalia chebula Retzius and Vitex negundo Linnaeus which are commonly used as home herbal remedies for the primary health care was carried out using Folin-Ciocalteu colorimetric method. Gallic acid was used as the standard for the estimation of phenolics. All the investigated plant extracts contained high amount of phenolics but the highest amount was detected in 70% acetone extract of B. ciliata (357.08 mg GAE/g sample) and the lowest amount was detected in 50% aqueous methanol extract of T. bellirica (108.69 mg GAE/g sample). The antioxidant activity of selected extracts was determined by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay. The extract having the highest phenolic content showed the lowest IC50 demonstrating the positive correlation between radical scavenging activity and total phenolic content. Nepal Journal of Science and Technology Vol. 14, No. 1 (2013) 95-102 DOI: http://dx.doi.org/10.3126/njst.v14i1.8928

Diocanol; one new phenol derivative isolated and characterized from Urtica dioica

One new benzene derivative Diocanol(1) was isolated from the ethyl acetate soluble portion of the whole plant of Urtica dioica along with known constituents β-amyrin (2), β-sitosterol (3), stigmasterol (4), and oleanolic acid (5). The structures of the isolated compounds were characterized based on 1H and 13C NMR spectra, including two-dimensional NMR techniques like COSY, HMQC, and HMBC and compared with the literature data.

A New Lupan type Triterpene Butilinol from Viburnum grandiflorum

The isolation and structural studies on the chemical constituents of Viburnum grandiflorum are described. The medicinal properties of the plant are also described. The mentholic extract was subjected to the preparative thin layer chromatography (PTLC) test experiments to investigate the isolation pattern. Based on the PTLC test experiments, the extract was subjected to the silica gel column chromatography. The column was eluted with increasing polarities of organic solvents. This afforded several fractions. The fractions were re- chromatographed on silica gel column to afford a new lupan type triterpene butilinol (1) with several known compounds i. e. oleanolic acid (2), ursolic acid (3), β-sitosterol (4), butilinic acid (5), butilin (6), α-amyrin (7) and germanicol (8). The compound (6) was not reported previously from the genus Viburnum. This therefore represents its first report from Viburnum grandiflorum. The compounds (2) and (4) have been previously reported from Viburnum pronifolium while the compounds (3) and (8) from Viburnum opulus and Viburnum erubescens, respectively. This represents the first report of the presence of these compounds in Viburnum grandiflorum. The structures of the above compounds were identified on the basis of spectral data (UV, IR, Mass, 1 NMR, 13 C-NMR) and literature evidences. The hexane and ethyl acetate soluble portions of the