Total MeOH Research Articles

In Vitro Acetylcholinesterase Inhibitory and Antioxidant Activity ofAlphonsea tonkinensisA.DC

Knowledge of the bioactivity of Alphonsea tonkinensis A.DC is limited. We have investigated the in vitro acetylcholinesterase inhibitory and antioxidant activities of extracts and pure compounds isolated from stems and leaves of this species collected from Dakrong district, Quang Tri Province, Vietnam. Extracts and isolated compounds were obtained by using an in-house extraction and chromatographic technique. The in vitro acetylcholinesterase inhibitory and antioxidant activities were evaluated using an Ellman test and 2,2-diphenyl-1-picryl-hydrazyl test, respectively. The total MeOH and CH2Cl2extracts, the MeOH portion of the CH2Cl2extract, pseudocolumbamine, and pseudopalmatine showed potential inhibitory activity against acetylcholinesterase with IC50values of 22.7, 32.9, 14.6, 18.9, and 8.6 μM, respectively. The aqueous phase (pH 9), MeOH portion of the CH2Cl2extract, and N- trans-feruloyltyramin exhibited significant antioxidant activities with IC50values of 24.5, 72.1, and 61.2 µM, respectively. This is the first study showing such bioactivities of various extracts obtained from A. tonkinensis.

Cell wall O-acetyl and methyl esterification patterns of leaves reflected in atmospheric emission signatures of acetic acid and methanol.

Plants emit high rates of methanol (meOH), generally assumed to derive from pectin demethylation, and this increases during abiotic stress. In contrast, less is known about the emission and source of acetic acid (AA). In this study, Populus trichocarpa (California poplar) leaves in different developmental stages were desiccated and quantified for total meOH and AA emissions together with bulk cell wall acetylation and methylation content. While young leaves showed high emissions of meOH (140 μmol m-2) and AA (42 μmol m-2), emissions were reduced in mature (meOH: 69%, AA: 60%) and old (meOH: 83%, AA: 76%) leaves. In contrast, the ratio of AA/meOH emissions increased with leaf development (young: 35%, mature: 43%, old: 82%), mimicking the pattern of O-acetyl/methyl ester ratios of leaf bulk cell walls (young: 35%, mature: 38%, old: 51%), which is driven by an increase in O-acetyl and decrease in methyl ester content with age. The results are consistent with meOH and AA emission sources from cell wall de-esterification, with young expanding tissues producing highly methylated pectin that is progressively demethyl-esterified. We highlight the quantification of AA/meOH emission ratios as a potential tool for rapid phenotype screening of structural carbohydrate esterification patterns.

Iridoids and other constituents from Cyperus rotundus L. rhizomes

On continuation of the phytochemical investigation of the MeOH extract of Cyperus rotundus L. Rhizomes, four compounds namely, methyl 3,4-dihydroxy benzoate (1), ipolamiide (2), 6β-hydroxyipolamiide (3), and rutin (4) were isolated. Their structures were established on the basis of 1D and 2D NMR spectroscopic analyses as well as ESIMS spectral data. Compounds 2 and 3 are reported here for the first time from the genus Cyperus and 1 for the first time from the plant. Co-treatment of CCl4 hepatic injured rats with the total MeOH extract (TME) and EtOAc fraction significantly restored the hepatic marker enzymes and total bilirubin to near-normal values compared to silymarin (reference drug).

Blepharisides A and B, new flavonol glycosides from Blepharis ciliaris growing in Saudi Arabia

Bio-guided fractionation of the total MeOH extract (TME) of the aerial parts of Blepharis ciliaris (L.) B.L. Burtt. (Acanthaceae) growing in Saudi Arabia was carried out to evaluate its hepatoprotective activity against CCl4-induced hepatotoxicity in rats. Successive chromatographic separations of the potent hepatoprotective n-BuOH fraction afforded two new flavonol glycosides, blepharisides A (1) and B (2), along with quercetin 3-O-rutinoside (3). Their structures were established by UV, IR, 1D, 2D NMR, and HRESIMS spectral data, in addition to comparison with literature data. Co-treatment of CCl4 hepatic injured rats with the total MeOH extract (TME) and its fractions significantly restored the hepatic marker enzymes and total bilirubin to near-normal values compared to silymarin (reference drug). The isolated compounds were evaluated for their antioxidant and anti-inflammatory activities. They displayed significant antioxidant activity (DPPH assay) in relation to propyl gallate (positive control) (% inhibition of 88.2, 87.9, and 74.2, respectively). Compounds 1 and 2 demonstrated anti-inflammatory effects in the carrageenin induced paw edema method at a dose of 10mg/kg.

Hypoestenonols A and B, new fusicoccane diterpenes from Hypoestes forskalei

Two new fusicoccane diterpenes hypoestenonols A (1) and B (2), along with two known compounds verticillarone (3) and hypoestenone (4) were isolated from the n-hexane fraction of the methanolic extract of the aerial parts of Hypoestes forskalei (Acanthaceae) growing in Saudi Arabia. The structures were established by UV, IR, HRESIMS, 1D (1H and 13C NMR) and 2D (1H1H COSY, HSQC, HMBC, and NOESY) NMR experiments, in addition to comparison with literature data. The total MeOH extract and isolated compounds were tested for their antiprotozoal and cytotoxic activities. The MeOH extract showed moderate activity against Plasmodium falciparum, Leishmania infantum, Trypanosoma cruzi, and Trypanosoma brucei with IC50 values of 8.8, 8.1, 9.1 and 8.1μg/mL without cytotoxicity (IC50 >64μg/mL on MRC5 cells). A very weak in vitro antiplasmodial effect was observed for hypoestenonol A (1) (IC50 18.9μM), verticillarone (3) (IC50 25.1μM), and hypoestenone (4) (IC50 16.7μM).

Stigmasterol Tetracosanoate, a New Stigmasterol Ester from the Egyptian Blepharis ciliaris.

A new stigmasterol ester: stigmasterol tetracosanoate (3), along with 7 compounds: β-sitosterol (1), stigmasterol (2), (2S,3S,4R)-2[(2'R)-2'-(hydroxyeicosanoyl amino) octadecane-1,3,4-triol (4), apigenin (5), β-sitosterol-3-O-β-D-glucopyranoside (6), stigmasterol-3-O-β-D-glucopyranose (7), and apigenin-7-O-β-D-glucopyranoside (8) were isolated from Blepharis ciliaris aerial parts. Compounds 1, 2, and 5-7 are reported here for the first time from the plant and 4 for the first time from the family. GCMS analysis revealed the presence of 45 fatty acids, 53 hydrocarbons, and 24 sterols. The different fractions exhibited mild cytotoxic in brine shrimp assay and anti-hyperglycaemic activities. The EtOAc fraction and TME (total MeOH extract) showed weak anti-malarial activity against P. falciparum. The CHCl3 fraction gave potent -anti-inflammatory activity compared with indomethacin.

New compounds from the Red Sea marine sponge Echinoclathria gibbosa

Three new compounds; β-sitosterol-3-O-(3Z)-pentacosenoate (1), 5α-pregna-3β-acetoxy-12β,16β-diol-20-one (2), and echinoclathriamide ((R)-2′-hydroxy-N-((2S,3S,4R)-1,3,4-trihydroxy-19-methylicosan-2-yl)heptadecanamide) (3), together with two known compounds; thymine (4) and uracil (5) were isolated from the EtOAc fraction of the Red Sea sponge Echinoclathria gibbosa. Their structures were unambiguously established on the basis of 1D and 2D NMR spectroscopy, in addition to mass spectrometry. The total MeOH extract (TME) and its fractions were evaluated for their antimicrobial, anti-inflammatory, antipyretic, and hepato-protective activities. The in vitro growth inhibitory activity of the isolated compounds was evaluated against three human cancer cell lines including the A549 non-small cell lung cancer (NSCLC), U373 glioblastoma (GBM), and PC-3 prostate cancer cell lines. Compound 1 showed weak activity against the three cancer cell lines.

Chemical constituents, anti-inflammatory, and antioxidant activities of Anisotes trisulcus

Anisotes trisulcus (Forssk.) Nees. (family Acanthaceae) aerial part is used in folk medicine in the Arabian peninsula for treatment of hepatic conditions. It showed different activities such as antibacterial, hepatoprotective, and cytotoxicity. It is a rich source of alkaloids and is used as an antidiabetic, bronchodilator, hypotensive, and local anesthetic. To the best of our knowledge, there is no report on the phenolic constituents of A. trisulcus. Therefore, this study aims to identify the constituents and establish antioxidant and anti-inflammatory activities of the total methanolic extract and different fractions. One new benzoyl flavonol: 7,8,3′-trihydroxy-5-methoxy-4′-benzoyl flavonol (5), along with eight known compounds: α-amyrin (1), β-sitosterol (2), stigmasterol (3), (2S,3S,4R)-2[(2′R)-2′-hydroxytetracosanoyl amino]-octadecane-1,3,4-triol (4), allopateuletin (6), veratric acid (7), vanillic acid (8), and β-sitosterol-3-O-β-d-glucopyranoside (9) were isolated from A. trisulcus aerial parts. Their structures were established by physical, chemical, and spectral data (UV, IR, MS, and 1D NMR), as well as comparison with authentic samples. The anti-inflammatory activity of the total methanolic extract and different fractions was evaluated using carrageenan-induced paw edema method at a dose of 400mg/kg. Also, the antioxidant activity was determined using DPPH assay at concentrations 0.25, 0.5, and 1mg/mL. The total MeOH extract and EtOAc fraction showed high antioxidant activity 75% and 68% (Conc. 1mg/mL), respectively while, the n-hexane and EtOAc fractions exhibited significant anti-inflammatory effects.

Ultrasound enhanced permeation of methanol into zebrafish, Danio rerio, embryos

Only vitrification is likely to result in successful cryopreservation of teleost embryos, requiring a high level of cryoprotectant penetration into the cell. This study determined the effect of ultrasound on methanol (MeOH) penetration in three types (dechorionated, soft chorion and intact embryos) and three stages (90% epiboly, bud and 4-somite stages) of zebrafish ( Danio rerio) embryos. Three types of embryos were first exposed to varying levels of methanol and optimized for survival. Dechorionated embryos were then exposed to 20%, soft chorion embryos were exposed to 30% and the intact embryos were exposed to 40% methanol for 3 min with or without ultrasound treatment. High performance liquid chromatography (HPLC) was used to measure methanol levels in the treated embryos. The total MeOH recovery from zebrafish embryo samples spiked with MeOH was 100.62 ± 3.99%. The overall penetration of methanol into zebrafish embryo ranged from 85.3 ± 8.1 μmol (in soft chorion embryos at 90% epiboly stage treated with ultrasound) to 3.1 ± 1.0 μmol (in dechorionated embryos at 90% epiboly stage treated without ultrasound). Ultrasound clearly helped to improve the permeation level of methanol into zebrafish embryos ( p < 0.05). A high level of methanol was found in soft chorion embryos. Dechorionated embryos had the highest relative difference in methanol levels between ultrasound and controls. Although, ultrasound significantly affected survival ( p < 0.05), survival was generally high (64 ± 2% to 100%). Even the highest level of methanol (85.3 ± 8.1 μmol) in the best treatment was far lower than that needed for vitrification. Further studies using novel and non-invasive methods are needed to achieve greater methanol penetration for successful vitrification.

Evaluation of Biological Activity of Turkish Plants. Rapid Screening for the Antimicrobial, Antioxidant, and Acetylcholinesterase Inhibitory Potential by TLC Bioautographic Methods

Using thin-layer chromatography (TLC) bioautography, a total of 58 extracts from various organs (aerial parts, leaves, flowers, fruits, roots) of 16 Turkish plants were tested for their antibacterial, antifungal, acetylcholinesterase inhibitory, antioxidant, and radical scavenging activities. The hexane, CHCl3/CH2Cl2, water, and total MeOH extracts were used. No activity was observed against two Gram-negative bacteria (Escherichia coli and Pseudomonas aureginosa) and the yeast Candida albicans. However, 23 plant extracts, mostly the CHCl3/CH2Cl2 and H2O-solubles, inhibited the growth of all five Gram-positive bacteria tested, Micrococcus luteus, Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Staphylococcus epidermidis. Of the active extracts, the CHCl3-soluble of the roots of Putoria calabrica (L. fil) DC (Rubiaceae) displayed the highest antibacterial potential. The majority of the CHCl3/CH2Cl2 crude extracts also appeared to inhibit acetylcholinesterase on TLC plates at 100 µg/spot concentration. Particularly active samples were the middle polarity extracts (CHCl3/CH2Cl2) of the leaves of Rhododendron smirnovii Trautv., R. ponticum L., and R. ungernii Trautv. (Ericaceae). β-Carotene, β-carotene/linoleic acid mixture, and 2,2-diphenyl-l-pieryhydrazyl (DPPH) solutions sprayed onto TLC plates were used for detecting antioxidant and radical scavenging properties of the crude extracts. Antioxidant and radical scavenging activities were found to be predominant in highly polar extracts. The water-solubles of all Rhododendron (Ericaceae) and Phlomis (Lamiaceae) species presented the most significant activity.

The constituents ofTaraxacum hallaisanensis roots

Three sesquiterpene lactone compounds, two novel(1β, 3β-dihydroxy-6β, 11β, 4α, 5α, 7αH-eudesm-12,6-olide-1-O-β-D-glucopyranoside, 1β, 3β-dihydroxy-6β, 4β, 5α, 7αH-eudesm-11 en-12,6-olide-1-O-β-D-glucopyranoside) and 1β, 3β-dihydroxy-6β, 11β, 4α, 5α, 7αH-eudesm-12,6-olide were isolated from the aqueous fraction of MeOH extract of the roots fromTaraxacum hallaisanensis (Compositae) employing Amberlite XAD-2, ODS-gel, silica gel and Sephadex LH-20 column chromatographics. Another known compound, (−)-epicatechin, was isolated from the aqueous fraction of the MeOH extract. The total MeOH extract also contained phytosterol and a mixture of β-amyrin acetate, α-amyrin acetate and lupeol acetate. Structures of isolated compounds were elucidated by spectroscopic parameters of IR, Mass,13C-NMR,1H-NMR,1H-1H COSY,13C-1H COSY and HMBC.

Changes in the degree of pectin demethylation during fermentation of fruit musts

AbstractMusts from apples, plums, black and red currants, pears, cherries and strawberries were subjected to fermentation. In the 84th h of fermentation a definite amount of exogenous pectin was added to some samples. Based on the determined content of methanol (MeOH) and its total presence, measured after alkaline hydrolysis, the degree of pectin demethylation (DDM) was calculated. Great variations of DDM as well as initial and total content of MeOH in particular musts were found. The highest DDM was found in a cherry must (83.4%) and red currant (72.4%). The remaining musts were demetylated in the range from 20 to 65%, except depectinized samples (72—98%). All the musts of 1986 displayed higher contents of total MeOH and lower degrees of final demethylations than those of 1984. It has been shown that endogenous pectin contained in musts is not fully demethylated, in spite of the fact that the solution is capable of further demethylation of exogenous pectin.