Evaluation of cytotoxic compounds from calligonum comosum L. growing in Egypt

Calligonum comosum (Polygonaceae), an Egyptian desert plant, was extracted and fractionated using petroleum ether, methylene chloride, and ethyl acetate. The total methanolic extract and other fractions were tested for their anticancer activity using Ehrlich ascites, brine shrimp and antioxidant assays. Ethyl acetate fraction proved to be the most active in all assays. Eight compounds were isolated, purified, and identified from this fraction as (+)-catechin (1), dehydrodicatechin A (2), kaempferol-3-O-rhamnopyranoside (3), quercitrin (quercetin-3-O-rhamnopyranoside) (4), beta-sitosterol-3-O-glucoside (5), isoquercitrin (quercetin-3-O-glucopyranoside) (6), kaempferol-3-O-glucuronide (7), and mequilianin (quercetin-3-O-glucuronide) (8). All isolated compounds were tested for their cytotoxicity and antioxidant activity. Compound 2 showed the best cytotoxic and antioxidant activity.     

An unusual C6-C6" linked flavonoid from Miconia cabucu (Melastomataceae)

Chromatographic fractionation of the methanolic extract from the leaves of Miconia cabucu Hoehne (Melastomataceae) afforded the first example of a C(6)-C(6") linked flavone dimer, 5-hydroxy-4',7-dimethoxyflavone-(6-C-6')-5'-hydroxy-3',4',7'-trimethoxyflavone as well as the known compounds, quercetin-3-O-alpha-l-rhamnopyranosyl-(2-->1)-O-beta-D-xylopyranoside, quercetin-3-O-alpha-l-rhamnopyranoside, myricetin-3-O-alpha-l-rhamnopyranoside, quercetin-3-O-beta-D-glucopyranoside, kaempferol-3-O-beta-D-(6'-coumaroyl)-glucopyranoside and gallic acid. Their chemical identities were established by application of NMR spectroscopic methods including 2D-NMR, as well as UV and ESI-MS analyses.     

Flavonoids from Achillea nobilis L

The detailed investigation of a methanolic extract of aerial parts of Achillea nobilis resulted in the isolation of 10 flavonoids. A new C-glycosylflavone, luteolin-6-C-apiofuranosyl-(1'-->2')-glucoside, was isolated besides orientin, isoorientin, vitexin, isoschaftoside, luteolin-7-O-beta-glucuronide, luteolin-4'-O-beta-glucoside and quercetin-3-O-methyl ether and two rare flavonolglycosides, quercetin-3-O-alpha-arabinosyl-(1'-->6')-glucoside and quercetin-3-O-methylether-7-O-beta-glucoside. The structures were established either by comparison with authentic substances or by UV, 1H NMR and 13C NMR spectroscopic methods including 2D-NMR techniques and ESI-MS.     

Polyphenolic compounds from flowers of Hibiscus rosa-sinensis Linn. and their inhibitory effect on alkaline phosphatase enzyme activity in vitro

Graded concentrations (0.1-100 mg/mL reaction mixture) of the methanolic extract of the flowers of Hibiscus rosa-sinensis Linn., its water-soluble fraction as well as compounds isolated from this fraction were tested for their inhibitory effect on alkaline phosphatase enzyme activity in vitro. Both the methanolic extract and its water-soluble fraction showed significant inhibitory effects on the enzyme activity in vitro. On screening the activity of the compounds isolated from the water-soluble fraction, its high inhibitory activity was attributed to the presence of quercetin-7-O-galactoside which showed a high potent inhibition of the enzyme activity reaching 100% at 100 mg/mL reaction mixture. Phytochemical investigations of the water-soluble fraction were also carried out and afforded ten polyphenolic compounds including two new natural compounds, namely kaempferol-7-O-[6'-O-p-hydroxybenzoyl-beta-D-glucosyl-(1-->6)-beta-D-glucopyranoside] and scutellarein-6-O-alpha-L-rhamnopyranoside-8-C-beta-D-glucopyranoside). The chemical structure of the isolated compounds was elucidated on the basis of chemical and spectral data.     

Bioactivity-guided isolation of the active compounds from Rosa nutkana and quantitative analysis of ascorbic acid by HPLC

Rosa nutkana Presl. (Rosaceae) is distributed abundantly throughout central and southern areas of British Columbia, Canada. Aboriginal people in the Pacific Northwest have traditionally used R. nutkana as a food, medicine, and source of cultural material. The methanolic extract of the fruits of R. nutkana was previously found to have inhibitory activity against methicillin-resistant Staphylococcus aureus (MRSA). In our study, bioactivity-guided fractionation of the methanol extract from R. nutkana led to the isolation of the following 10 compounds: (i) tormentic acid, (ii) euscaphic acid, (iii) ursolic acid, (iv) maslinic acid, (v) quercetin, (vi) catechin gallate, (vii) quercetin-3-O-glucoside, (viii) 1,2,3,4,6-penta-O-galloyl-beta-D-glucoside, (ix) L-ascorbic acid (vitamin C), and (x) 1,6-digalloyl-beta-D-glucoside. Structures were elucidated by ultraviolet, infrared, mass spectrometry, and nuclear magnetic resonance data, as well as by comparison with those of the literature. The compounds quercetin, catechin gallate, quercetin-3-O-glucoside, 1,2,3,4,6-penta-O-galloyl-beta-D-glucoside, and 1,6-digalloyl-beta-D-glucoside exhibited weak antibacterial activity against MRSA. Our research demonstrates the value of traditional knowledge held by Aboriginal people in the Pacific Northwest with respect to uses of R. nutkana. Some described uses in the ethnobotanical literature correspond to activities observed under laboratory conditions. Further work on British Columbia Rosa spp. may contribute to identifying other potential therapeutic uses.     

番石榴叶化学成分分离鉴定及psiguadial D的抗癌活性研究

本文对番石榴Psidium guajava叶的化学成分进行分离鉴定并对部分化合物抗肿瘤活性进行评价。采用正相硅胶、Sephadex LH-20、MCI柱色谱和制备液相色谱等方法对番石榴叶甲醇提取物的化学成分进行分离纯化,运用现代波谱技术鉴定了13个化合物,分别为4,5-diepipsidial A(1)、psidial A(2)、guajadial(3)、psiguadial A(4)、psiguadial D(5)、α-生育酚(6)、亚油酸(7)、槲皮素-3-O-β-D-吡喃木糖苷(8)、槲皮素-3-O-α-L-吡喃阿拉伯糖苷(9)、槲皮素-3-O-α-L-呋喃阿拉伯糖苷(10)、β-谷甾醇(11)、儿茶素(12)、没食子酸(13)。其中化合物1为首次从天然产物中分离得到。化合物5对人体肿瘤细胞株HL-60、SMMC-7721、A-549、MCF-7、SW-480均显示出较为明显的抑制活性,半数抑制浓度为4.08~11.23μmol/L。     

Hepatoprotective activity of LC–ESI-MS standardized Iris spuria rhizome extract on its main bioactive constituents

The study was designed to evaluate the hepatoprotective activity of Iris spuria against paracetamol induced toxicity at two different doses 100 and 200 mg/kg. The extract showed significant protective activity (p > 0.01) at both the doses in dose dependent manner. Administration of the plant extract restored the paracetamol induced elevated levels of serum marker and distorted hepatic tissue architecture. The lipid peroxides (LPO) and glutathione (GSH) levels were also restored towards normal in liver tissue significantly. The main chemical constituents of the extract identified by the liquid chromatography–tandem mass spectrometry (LC–ESI-MSMS) were found to be flavones and isoflavonoids. Tectoridin and iristectorigenin A were the principal compounds present in the methanolic extract of Iris spuria.     

Biochemical constituents of a wild strain of Schizophyllum commune isolated from Achanakmar-Amarkantak Biosphere Reserve (ABR), India

A wild strain of Schizophyllum commune (MTCC 9670) isolated from Achanakmar-Amarkantak Biosphere Reserve of Central India was evaluated for the production of bioactive compounds. The chemical constituents of wild and in vitro grown cultures were compared. Under optimized conditions, different organic and aqueous extracts from mycelia and fruiting bodies were used to extract chemical components from the cultures grown in vitro. The gas chromatography combined wih mass spectrometry analysis of extracts identified two phenolic compounds, namely Phenyl benzoate (C₁₃H₁₀O₂) and 4-(phenyl methoxy) phenol (C₁₃H₁₂O₂) in the ethanolic extract of in vitro grown fruiting bodies and one antibacterial compound Pyrrolo (1, 2-a) piperazine-3, 6-dione (C₇H₁₀O₂N₂) in the methanolic extract of mycelia. High-performance liquid chromatography analysis revealed that the gallic acid and l-ascorbic acid were identifiable antioxidant components in the extracts possessing high free radical scavenging activity. The findings suggest that the wild strain of S. commune may serve as the source of novel bioactive compounds with effective antimicrobial and antioxidant activities.     

Comprehensive separation and identification of chemical constituents from Apocynum venetum leaves by high-performance counter-current chromatography and high performance liquid chromatography coupled with mass spectrometry

High-performance counter-current chromatography (HPCCC) and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was efficiently utilized for the separation and identification of the chemical components with a wide range of polarity from the mixed extract of Chinese medicinal herb Apocynum venetum. For HPCCC separation, four sets of solvent systems, n-hexane-ethyl acetate-acetonitrile-water (1.5:3.5:2:4.5, v:v:v:v), ethyl acetate-methanol-water (5:2:5, v:v:v) and n-butanol-methanol-water (5:1:5, v:v:v) were used for the one-step separation by four stages. The HPCCC separation was initiated by filling the column with the lower phase of n-hexane-ethyl acetate-acetonitrile-water (1.5:3.5:2:5, v:v:v:v) as a stationary phase followed by elution with the upper phase of n-hexane-ethyl acetate-acetonitrile-water (1.5:3.5:2:5, v:v:v:v) to separate the hydrophobic compounds (tail to head). Then the mobile phase was switched to the upper phase of ethyl acetate-acetonitrile-water (5:3:7, v:v:v) to eluted the moderate hydrophobic compounds, then the mobile phase was switched to the upper phase of ethyl acetate-methanol-water (5:2:5, v:v:v) to eluted the moderate hydrophilic compounds, and finally the hydrophilic compounds still retained in the column was eluted by the upper phase of n-butanol-methanol-water (5:1:5, v:v:v). A total of 16 named compounds including adhyperforin, hyperforin, amentoflavone, biapigenin, quercetin, avicularin, acetylated isoquercetin, acetylated hyperoside, astragalin, trifolin, isoquercetin, hyperoside, querciturone, rutin, chlorogenic acid and quercetin-3-O-β-D-glucosyl-β-D-glucopyranoside were successfully separated via the four sets of solvent systems in one step operation for 130 min. The compounds separated by HPCCC were identified by comparing with mixed standards data of HPLC-MS as well as NMR data.     

西南獐芽菜中一个新的裂环烯醚萜苷

为研究西南獐芽菜中的水溶性化学成分,该研究将西南獐牙菜80%甲醇提取物经二氯甲烷萃取后,水溶部分通过柱色谱及高效液相制备色谱分离纯化,结合波谱分析鉴定结构。结果表明:从西南獐芽菜甲醇提取物中分离并鉴定得到了一个新裂环烯醚萜苷,命名为西南獐牙菜内酯C。     

Phytochemical Profiling and Compound Isolation of Cissus rhombifolia Vahl. Leaves Aqueous Methanolic Extract with the Evaluation of Its Anti-Inflammatory Effect Using Lipopolysaccharide-Induced Inflammation in RAW 264.7 Cells

The inflammatory disorders represent a serious health issue. Certain Cissus species possess anti-inflammatory effect. Cissus rhombifolia Vahl. leaves’ anti-inflammatory activities and phytoconstituents are poorly characterized. In this study, 38 constituents were tentatively characterized in Cissus rhombifolia Vahl. leaves’ aqueous methanolic extract (CRLE) using high-performance liquid chromatography combined with mass spectrometry (HPLC/MS) and Proton Nuclear Magnetic Resonance (¹H-NMR). Myricetin, β-amyrin, and alliospiroside A, were isolated from CRLE using column chromatography. The anti-inflammatory effect of CRLE and its isolated compounds were studied in lipopolysaccharide (LPS)-induced RAW 264.7 cells. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay) was used to assess how CRLE and its isolated compounds affected cell viability. Further, its effects on the production of intracellular NO, and inflammatory cytokines cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6) were assessed by the Griess test, and cytokine enzyme-linked immunosorbent assays, respectively. CRLE and its isolated compounds, myricetin, β-amyrin, and alliospiroside A decreased the NO production. Western blotting was performed to assess the protein expression levels of the inflammatory cytokines inducible nitric oxide synthase (iNOS). Alliospiroside A downregulated IL-6, TNF-α, and COX-2 and inhibited the expression of iNOS. CRLE and its compounds represent effective alternative candidate to treat inflammatory diseases.     

GC/MS and LC/MS Phytochemical Analysis of Vigna unguiculata L. Walp Pod

Vigna unguiculata (L. Walp) or Cowpea pod methanolic extracts phytochemical analysis, total phenolic content (TPC), and secondary metabolite profiling were determined using gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-mass spectrometry (LC/MS) analysis. GC/MS analysis revealed twenty compounds in the extract, while LC/MS analysis identified twenty-four compounds. GC/MS chromatogram analysis suggested the presence of opioid α-N-Normethadol a major constituent found in methanolic extract and fatty acid esters carotenoid is found second major constituent. LC/MS chromatogram and the mass spectral analysis demonstrated the presence of flavonoids, carotenoids, and alkaloids as major phytochemicals. We investigated the antibacterial, anti-fungal, and anti-oxidant activity of pod methanolic extract. The extract was found equally effective against E. coli, S. pyogenes, and P. aeruginosa with MIC 100 μg/mL similar to the standard Ampicillin (MIC 100 μg/mL). C. albicans were found to be most susceptible to Vign unguiculata pods methanolic extract with a MIC of 250 μg/mL. The pod extract showed significant DPPH scavenging activity (IC₅₀=78.38±0.15) which suggests its antioxidant potential.     

Hairy roots of Dracocephalum moldavica: rosmarinic acid content and antioxidant potential

Hairy roots of Dracocephalum moldavica L. were induced using Agrobacterium rhizogenes strain A4. Transformed roots were obtained from shoot explants with low transformation frequency of up to 3 %. The effects of different liquid media: Murashige and Skoog (MS), Gamborg et al. (B5) and Woody Plant (WP) with full- and half-strength (½MS, ½B5, ½WP), on biomass accumulation and rosmarinic acid (RA) content were investigated. The hairy roots were cultured in photoperiod (16 h light/8 h dark) and darkness. Biomass of D. moldavica hairy roots was the highest (7.23 g flask^?1 of fresh weight and 0.89 g flask^?1 of dry weight) in the cultures grown in WP medium under periodic light. Ultra performance liquid chromatography analysis revealed the highest RA content (78 mg g^?1 dry wt) in roots cultured in ½B5 medium under photoperiod conditions. It was about tenfold higher compared to roots of field-grown mother plants. Antioxidant activities and total phenolic contents of methanolic extracts of D. moldavica hairy roots cultured in ½B5 and WP media under photoperiod and darkness and roots of field grown plants were compared. All extracts were investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging and phosphomolybdenum reduction assays. Total phenolic contents were estimated by the Folin–Ciocalteu method. The methanolic extract of D. moldavica hairy roots grown in ½B5 medium under photoperiod possessed the strongest effects on reducing Mo and DPPH radical scavenging. The activities were significantly higher (p ≤ 0.05) than those of methanolic extract of roots of intact plants grown in the field. The most active methanolic extract of hairy roots was characterized by the highest level of rosmarinic acid and total content of phenolic compounds.     

Isolation and identification of a phenolic antioxidant from Aloe barbadensis

A potent antioxidative compound has been isolated from a methanolic extract of Aloe barbadensis Miller using a combination of column and thin-layer chromatography. The antioxidant activity of this substance was similar to that of alpha-tocopherol as assessed in vitro using rat brain homogenates. On the basis of electrospray ionization and electron-impact ionization mass spectra in combination with reversed-phase, high-performance liquid chromatographic behavior, this compound has been identified as 8-C-beta-D-[2-O-(E)-coumaroyl]glucopyranosyl-2-[2-hydroxy]-propyl-7-methoxy-5-methylchromone.     

相似蜂海绵相关真菌杂色曲霉F62活性代谢产物研究

研究了1株相似蜂海绵相关真菌F62的活性代谢产物,经分子系统学分析表明该真菌属于杂色曲霉。将F62菌株用大米固体培养基在室温（约25℃）下静置培养45d,经乙酸乙酯超声萃取得到粗提物。通过硅胶柱层析、Sephadex LH-20凝胶柱层析和HPLC等色谱手段,分离得到了6个化合物。通过核磁共振、质谱等波谱分析手段可鉴定出其结构分别为Alantrypinone（1）、洛伐他汀（2）、甲酯型莫纳克林K（3）、土曲霉酮（4）、土震素B（5）和麦角甾醇（6）。化合物1系首次从该属真菌中分离得到,化合物2－5为首次从该种真菌中分离得到。首次对化合物3的碳谱数据进行报道。初步的药理研究表明,化合物4具有体外抗炎活性。     

Caesalpinia palmeri: First Report on the Phenolic Compounds Profile,Antioxidant and Cytotoxicity Effect

This study aimed to determine the phenolic compounds profile, antioxidant potential and cytotoxicity of extracts and fractions of Caesalpinia palmeri. Methanolic extracts were generated from C. palmeri berries, stems and flowers. The latter was subjected to liquid-liquid partition, obtaining hexane, ethyl acetate and residues fractions. Results showed that the flower extract and ethyl acetate fraction had a larger concentration of phenolic compounds (148.9 and 307.9 mg GAE/g, respectively), being ellagic acid (6233.57 and 19550.08 μg/g, respectively), quercetin-3-β-glycoside (3023.85 and 8952.55 μg/g, respectively) and gallic acid (2212.98 and 8422.34 μg/g, respectively) the most abundant compounds. Flower extract and ethyl acetate fraction also presented the highest antioxidant capacity on all tested methods (DPPH, ABTS, ORAC and FRAP) and low cytotoxicity against ARPE-19 cells (IC₅₀ >170 μg/mL). C. palmeri possessed high antioxidant potential, associated with the presence of phenolic compounds and low cytotoxicity, suggesting that they could represent an option to counter oxidative stress.     

Rapid separation of triterpenoids from Neem seed extracts

A dichloromethane extract of the triterpenoids from seeds of the Neem tree (Azadirachta indica) can be fractionated rapidly and with economic use of solvents using the Biotage™ flash chromatography system. The fractions can be analysed by thin layer chromatography (TLC), or quantitatively, by supercritical fluid chromatography or high performance liquid chromatography (HPLC), for the determination of 11 tetranortriterpenoids, including the natural insecticide azadirachtin. After a second pass through the Biotage flash column, pure compounds, including azadirachtin and salannin can be obtained directly, although these compounds comprise less than 0.3% of the total mass of the seeds extracted and are present in a complex mixture of similar substances. Copyright © 1999 John Wiley & Sons, Ltd.     

Protective effects of an extract from Citrus bergamia against inflammatory injury in interferon-γ and histamine exposed human keratinocytes

AimsThe present work evaluated the anti‐inflammatory/antioxidant activity of a well characterized extract from Citrus bergamia Risso and Poiteau (CBE), containing neoeriocitrin, naringin, neohesperidin and other flavonoids, on human NCTC 2544 keratinocytes treated with interferon-gamma (IFN-γ) and histamine (H).Main methodsHigh performance liquid chromatography (HPLC) coupled with diode array detectors was used to characterize and quantify phenolic compounds in CBE. Anti‐inflammatory/antioxidant ability on keratinocytes was determined through evaluation of inter-cellular adhesion molecule-1 (ICAM-1) and inducible nitric oxide synthase (iNOS) expression by Western blot, production of nitric oxide (NO) with Griess reagent and concentration of reactive oxygen species (ROS) by fluorescent quantitative analysis with 2′,7′-dichlorfluorescein-diacetate (DCFH-DA). Cell viability was assessed using 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Antioxidant activity was also measured by oxygen radical absorbance capacity (ORAC) assay. Glycosaminoglycans (GAGs) were quantified using 1.9-dimethyl methylene blue (DMB).Key findingsCBE exhibited high antioxidant activity confirmed by elevated ORAC values related to high capacity in oxygen radical scavenging. The assays on keratinocytes demonstrated that CBE does not inhibit cell proliferation and is shown to significantly reduce dose-dependently ICAM-1, iNOS, NO, ROS and GAG production in cells exposed to IFN-γ and H.SignificanceOur study demonstrates that the pools of compounds of an extract from C. bergamia efficiently block the proinflammatory actions induced by IFN-γ and H on human keratinocytes. CBE may be used for topic employment in some inflammatory diseases of the skin and to represent an important opportunity for the essential oil processing industries.     

Identification of 1-lysophosphatidylethanolamine (C(16:1)) as an antimicrobial compound in the housefly, Musca domestica

We observed that a methanolic whole body extract of uninfected last instar larvae of the housefly, Musca domestica, displayed antifungal and antibacterial activity. We have further purified this extract to a single active fraction using reversed phase high performance liquid chromatography. The pure fraction inhibited growth of the Gram-positive bacteria Bacillus thuringiensis and the yeast Saccharomyces cerevisiae, but not the Gram-negative bacteria Escherichia coli. The active compound was determined to have a molecular mass of 451.2 Da. Further analysis by nuclear magnetic resonance identified the substance as mono-unsaturated 1-lysophosphatidylethanolamine (C(16:1)) (1-LPE). The structurally different and more common 2-LPE have been described as mediators of the antimicrobial activity of rimenophenazine antibiotic agents (Van Rensburg et al., 1992). Our results suggest that the isolated 1-LPE displays a higher activity in comparison, possibly based on structure-specific differences in activity.     

Major constituents and cytotoxic effects of Ajuga chamaecistus ssp. tomentella

The n-butanolic fraction of a methanolic extract (80%) from aerial parts of Ajuga chamaecistus ssp. tomentella was analysed using different chromatographic methods. Column (CC) and high-performance liquid chromatography (HPLC) were used for isolation and purification. 13C, H NMR, H-H COSY, HSQC, HMBC, and ESI-MS were employed for identification of the compounds isolated from this fraction. The structures of the compounds were determined to be cis-melilotoside (1), trans-melilotoside (2), lavandulifolioside (3), 20-hydroxyecdysone (4), leonoside B (5), martynoside (6), ajugalactone (7), makisterone A (8), and 24-dehydroprecyasterone (9). This is the first report on the presence of cis- and trans-melilotoside in Ajuga species. Cytotoxic evaluation of the n-butanolic fraction, cis- and trans-melilotoside against cancer (T47D, HT-29, and Caco-2) and normal (NIH 3T3) cell lines by the mitochondrial tetrazolium test (MTT) showed no cytotoxic effects up to 400 microg/mL. The results of this study suggest that melilotoside, phenylethyl glycosides, and phytoecdysteroids are the main constituents of the n-butanolic fraction of Ajuga chamaecistus ssp. tomentella.