Organic-soluble Extracts Research Articles

Bioactive compounds from Matricaria chamomilla: structure identification, in vitro antiproliferative, antimigratory, antiangiogenic, and antiadenoviral activities.

During our exploring the anticancer activity of some medicinal plants and their major metabolites, the aerial parts of the Egyptian Matricaria chamomilla (flowers and stems) were studied. GC-MS analysis of the organic soluble extracts of the flowers and stems fractions revealed the presence of 43 and 45 compounds, respectively. Individual chromatographic purification of the flowers and stems' extracts afforded three major compounds. Structures of these compounds were identified by 1D- and 2D-NMR and HRESI-MS spectroscopic data as bisabolol oxide A (1) and (E)-tonghaosu (2) (as mixture of ratio 2:1) from the flowers extract, meanwhile apigenin-7-β-d-glucoside (3) from the stems fraction. Biologically, the chamomile extracts announced significant antiproliferative activities exceeded in potency by ∼1.5 fold in case of the stem, recording GI50 13.16 and 17.04μg/mL against Caco-2 and MCF-7, respectively. Both fractions were approximately equipotent against the migration of the same cell type down to 10μg/mL together, compounds 1, 2 but not 3, showed considerable growth inhibition of the same cells at GI50 13.36 and 11.83μg/mL, respectively. Interestingly, they were able to suppress Caco-2 colon cancer cells migration at 5.8μg/mL and potently inactivate the VEGFR2 angiogenic enzyme (1.5-fold relative to sorafenib. The obtained compounds and corresponding chamomile extracts were evaluated against Adeno-7 virus, revealing that both chamomiles' extracts (flowers and stems) and their corresponding obtained compounds (1-3) were potent in their depletion to the Adeno 7 infectivity titer, however, the flower extract and compounds 1-2 were more effective than those of the stem extract and its end-product (3).

A study of cytotoxicity and genotoxicity of particulate matter (PM2.5) in human lung epithelial cells (A549)

Exposure to airborne particulate matter (PM2.5), a PM with an aerodynamic diameter of less than 2.5 μm, is known to be associated with a variety of adverse health effects, particularly related to the respiratory system. However, the molecular mechanisms involved in fine PM toxicity are still not well-characterized. In this study, we estimate pulmonary toxic mechanism using two types (water soluble extract, WPM2.5, and organic soluble extract, O-PM2.5) of PM2.5 on human lung epithelial cells (A549). Samples were collected using a high-volume air sampler. Each sample was divided into two groups by its own types (water soluble extract, W-PM2.5, and organic soluble extract, O-PM2.5). In the present study, two types of PM2.5-induced cytotoxic and genotoxic effects and expression of toxicity-related genes were evaluated using human lung epithelial cells (A549). Also, the production of intracellular reactive oxygen species was measured to investigate the mechanism of cell death induced by PM2.5 Both W-PM2.5 and O-PM2.5 exposures significantly reduced the viability of A549 cells in a dosedependent manner, and expression of 17 cell deathrelated genes were significantly regulated in the PM2.5 exposure group. Exposure of PM2.5 significantly induced the production of ROS. Further, data obtained from the Comet assay indicated that two extracts of PM2.5 caused DNA damage in A549 cells in a dose-dependent manner. Our study suggests that ROS-mediated DNA damage may play a major role in PM2.5-induced cell death. This finding represents the basis for further studies addressing the pathophysiological mechanisms of PM2.5 exposure.

Further spectral and chromatographic studies of ambergris

Jetsam ambergris, found washed ashore on beaches, is an environmentally modified form of a natural product of Sperm whales which sometimes develops a pleasant odour. Odorous samples have proved valuable in perfumery. Identification of jetsam ambergris by analysis of organic-soluble extracts by Fourier transform infra-red spectroscopy (FTIR) and of derivatised samples by gas chromatography-mass spectrometry (GC-MS) has already been shown. Here, we describe a different method, in which characteristic alkenic protons and carbon atoms of the major constituent ambrein, were identified in whole extracts using nuclear magnetic resonance spectroscopy (NMR). The advantages of employing NMR spectroscopy included rapidity, reduced losses of volatiles compared to GC-MS and detection of non-GC amenable constituents. However, the identities and quantities of co-occurring individual components (e.g. steroids) could not easily be assigned in the unfractionated extracts by NMR spectroscopy, whereas they were by GC-MS, so an approach combining FTIR, GC-MS and NMR spectroscopic methods is advocated.

In Vitro Anthelmintic Activity Evaluation of the Aerial Part of Ruellia tuberosa Linn. Against Eudrilus eugeniae

The ever growing problems caused by perpetual helminth infections especially in third world countries has paved way for scientists to continually search for potential sources of alternatives. Ruellia tuberosa Linn., locally known as “cracker plant”, is a ubiquitous Philippine medicinal that is traditionally used as remedy for a variety of diseases. In the light of plant-based anthelmintic drug development, the present study was conducted to evaluate the anthelmintic activities of the decoction (RTD), crude ethanol (RTE), chloroform-soluble (RTC) and hexane-soluble (RTH) extracts and its fractions from the aerial part of R. tuberosa Linn. against Eudrilus eugeniae (African night crawler earthworms).Anthelmintic assay was employed at various concentrations in which the activity of the test samples was determined by correlation with time. Results showed that in vitro anthelmintic assay of crude ethanol extract relatively gave better results with respect to paralysis and death of earthworms compared to its decoction counterpart. Meanwhile, both organic soluble extracts exhibited notably better anthelmintic activities and were faster to bring about paralysis on the worms as compared to the aqueous and alcoholic extracts. Furthermore, all fractions of the nonpolar and mid-polar extracts showed significantly higher anthelmintic activities compared to its mother extracts (RTH and RTC). Additionally, among all organic fractions tested, RTC4 showed fairly high mortality rates incurred on the test organisms in which similar result was obtained in comparison for its two parent crude extracts RTC and RTE. A concentration-dependent toxicity relationship was also observed during the conduct of all bioassays. Subsequently, among all extracts and fractions tested, RTC4 gave fastest mean time in paralyzing and killing the earthworms (34.42±4.00min and 36.11±4.32min), thus, exhibited best anthelmintic activity that is remarkably comparable to the positive control, mebendazole (54.58±2.17min and 76.49±12.60min). Considering the results from the mid-polar extracts of the plant sample for the given assay, R. tuberosa Linn. showed great potential for plant-derived anthelmintics.

Extraction method for manipulation of water- and organic-soluble extracts of PM2.5 in Korean winter season and its chemical composition

Particulate matter (PM) air pollution is a significant environmental issue. PM is divided into three main fractions based on its size: coarse (2.5–10 μm), fine (0.1–2.5 μm), and ultrafine fractions (<0.1 μm). Fine particles and ultrafine fractions which are designated as particulate matter 2.5 (PM2.5) is small enough to reach alveoli in the lungs. Although the correlation between the chemical composition of PM2.5 and its toxic mechanisms has not been elucidated, several studies of PM2.5 have found evidence that ambient air PM2.5 is associated with adverse health effects. In the present study, the chemical compositions of water- and organic-soluble extracts of PM2.5 from 50 sample filters collected in the winter in the middle part of Seoul were measured. The water-soluble extract was analyzed to determine its contents of inorganic ions and metals using ion chromatography, atomic absorption spectrometry, and inductively coupled plasma-atomic emission spectrometry. The main inorganic ions were Cl−, NO3−, SO42−, and NH4+, with a total quantity in the range of 20.02 to 40.22 μg/m3 in ambient air. In a sample of all five groups, Na, K, Ca, Mg, Cd, Cr, Cu, Fe, Mn, Ni, V, and Zn together accounted for 902.14 to 1631.25 ng/m3 in ambient air. The concentrations of thirteen polycyclic aromatic hydrocarbons (PAHs) in the organic-soluble extract were analyzed using gas chromatography/time of flight-mass spectrometry. The thirteen PAHs combined constituted 6.24–76.05 ng/m3 in ambient air. In this study, we report the chemical composition of PM2.5 during the winter season in Seoul and suggest using a high-volume air sampler to create water- and organic-soluble extracts from large amounts of PM2.5, which could be used to conduct in vitro and in vivo experiments, including studying genomic changes following PM exposure.

Age-related accumulation of 3-nitrotyrosine and nitro-A2E in human Bruch's membrane

Age-related macular degeneration (AMD) is a disease leading to severe visual loss and legal blindness in the elderly population. The pathophysiology of AMD is complex and may include genetic predispositions, accumulation of lipofuscin and drusen, local inflammation and neovascularization. Recently four independent research groups have identified a commonly inherited variant (Y402H) of the complement factor H gene in the genome from different groups of AMD patients. The Y402H variant of CFH significantly increases the risk of AMD and links the genetics of the disease with inflammation. During inflammation there is activation of inducible nitric oxide synthase and release of nitric oxide, which in principal could lead to non-enzymatic nitration within extracellular deposits and/or intrinsic extracellular matrix protein components of human Bruch's membrane. We have identified two biomarkers for non-enzymatic nitration in aged human Bruch's membrane, indicative of inflammation, that include 3-nitrotyrosine identified in Bruch's membrane preparations and nitrated A2E from the lipid soluble extract of the Bruch's membrane preparation. Approximately 30-40 times more A2E is observed in samples of the organic soluble extract of lipofuscin compared to the extract of Bruch's membrane. It is of interest to note that although A2E is a major constituent of RPE lipofuscin, nitrated A2E could not be detected in RPE extracts. We show here that nitro-A2E is a specific biomarker of nitrosative stress in Bruch's membrane and its concentration correlates directly with tissue age.

Micronuclei induced by airborne particulate matter from Mexico City

Particulate air pollution is an important environmental health risk. In the present study, we have investigated the ability of chemically characterized water and organic-soluble extracts of PM 10 from two different regions of Mexico City to induce micronuclei in a human epithelial cell line. We also evaluated the association between the chemical characteristics of the PM and its genotoxicity. The airborne particulate samples were collected from an industrial and a residential region; a Hi-Vol air sampler was used to collect PM 10 on glass fiber filters. PM mass was determined by gravimetric analysis of the filters. One section of each PM 10 filter was agitated either with deionized water to extract water-soluble compounds or with dichloromethane to prepare organic-soluble compounds. The chemical composition of the extracts was determined by ion and gas chromatography and atomic adsorption spectroscopy. A549-human alveolar epithelial cells were exposed to different concentrations of PM 10 extracts and the cytokinesis blocked micronucleus assay was performed to measure DNA damage. Even though the industrial region had a higher PM concentration, higher amounts of metals and PAHs were found in the residential area. Both industrial and residential extracts induced a significant concentration-related increase in the micronuclei frequency. The PM 10 water-soluble industrial extract induced significantly more micronuclei than the one of the residential region; inversely, the organic residential extract induced more micronuclei than the one from the industrial region. The association between the induction of micronuclei and the chemical components obtained by the comparative analysis of standardized regression coefficients showed that cadmium and PAHs were significantly associated with micronuclei induction. Data indicate that water-soluble metals and the organic-soluble fraction of PM 10 are both important in the production of micronuclei. Effects observed, point to the risk of PM exposure and shows the need of integrative studies.

Origin and distribution of biomarkers in the sulphur rich Utrillas coal basin – Teruel mining district – Spain

The Utrillas coal facies are located in the Maestrazgo basin in NE Spain. This mining district of Teruel contains sub-bituminous deposits from the Middle Albian (Lower Cretaceous 105 Ma) in areas near a delta estuary with abundant sulphur. The high sulphur content is due to an influx of sulphate caused by the geological recycling of Triassic gypsum from the catchment area into the delta estuary. In some outcrops, the weathered coal reveals leonardite deposits. The depositional environment of the basin originated coals, some of which are currently mined. The organic matter of the coals has been the object of scattered reports. Studies have focused on bulk pyrolysis parameters and microscopic observation in Utrillas samples, as well as the inorganic and insoluble organic fraction. We analysed the organic soluble extract of the Utrillas coals using GC–MS in order to characterize their aliphatic, aromatic and organosulphur compounds. The biomarker distribution allowed us to recognize different inputs, assess their depositional palaeoenvironment and finally determine their degree of maturity. In particular, homologous series of hopanes related to eubacteria were present. Biomarkers characteristic of higher plant inputs were also widely distributed (e.g. phyllocladane or C 29 steranes). The presence of linear alkylbenzenes allowed us to recognize the palaeodepositional reducing environments where they were deposited. Specifically, thienylhopanes were associated with sulphur-reducing environments. Finally, the abundance of unsaturated biomarkers such as diacholestenes indicated low-maturity coals. Various aromatic ratios such as the methylphenanthrene index also suggested diagenesis in the initial stage.

Characterization of an anti-tuberculosis resin glycoside from the prairie medicinal plant Ipomoea leptophylla.

The organic soluble extract from the leaves of the native North American prairie plant Ipomoea leptophylla (big root morning glory) showed in vitro activity against M. tuberculosis. Bioassay-guided fractionation of this extract resulted in the identification of two new resin glycosides (6, 7). Base-catalyzed hydrolysis of these glycosides gave operculinic acid (1) as the glycosidic acid component as well as trans-cinnamic acid, propanoic acid, and lauric acid. The complete structure elucidation was accomplished through derivatization, 1D and 2D NMR spectroscopy (TOCSY, ROESY, HSQC, HMBC), and MS/MS experiments on 6 and 7 as well as the permethylated derivative 8.

Isolation of an anti-HIV diprenylated bibenzyl from Glycyrrhiza lepidota

The organic soluble extract from the leaves of Glycyrrhiza lepidota showed moderate activity in the US National Cancer Institute in vitro anti-HIV-1 bioassay. Chromatographic separation of this extract resulted in the identification of a new diprenylated bibenzyl as the compound responsible for the observed anti-viral activity. Extensive spectroscopic experiments provide the complete 1H NMR and 13C NMR spectral assignments to support the proposed structure. Known compounds glepidotin B and glepidotin A were also isolated from the extract and shown to be inactive in the anti-viral assay.

New insecticidal rocaglamide derivatives and related compounds from Aglaia oligophylla.

Organic-soluble extracts of the twigs of Aglaia oligophylla collected in Vietnam yielded four insecticidal cyclopentatetrahydrobenzofurans of the rocaglamide type including one new natural product (compound 4). Moreover, two cyclopentatetrahydrobenzopyran derivatives, belonging to the aglain and aglaforbesin types, respectively, were also isolated. The aglaforbesin derivative 6 proved likewise to be a new natural product. All isolated rocaglamide, aglain, and aglaforbesin derivatives have a characteristic methylenedioxy substituent linked to C-6 and C-7 or to C-7 and C-8, respectively. Structure elucidation of the new natural products and the determination of the absolute configuration of compound 1 by calculation of its CD spectrum with molecular dynamics simulation are described. All isolated rocaglamide derivatives exhibited strong insecticidal activity toward neonate larvae of the polyphageous pest insect Spodoptera littoralis when incorporated into an artificial diet, with LC(50) values varying between 2.15 and 6.52 ppm.

Plant Secondary Metabolites as Potential Anticancer Agents and Cancer Chemopreventives

A. Douglas KinghornProgram for Collaborative Research in the Pharmaceutical Sciences and Department of MedicinalChemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, 833 SouthWood Street, Chicago, IL 60612, U.S.ATel.: (312) 996-0914, Fax: (312) 996-7107, E-mail: kinghorn@uic.eduThere is considerable interest in the screening of plant and other natural product extracts in moderndrug discovery programs, since structurally novel chemotypes with potent and selective biological ac-tivity may be obtained [1-4]. A consideration of biological activity in addition to the isolation andstructure elucidation stages in a phytochemical investigation may add a great deal to the overall scien-tific significance of the work. Phytochemists may gain considerable information by using panels ofsimple bioassays and/or more specialized in vitro bioassays to follow each step of a purification proce-dure [3]. In the following paragraphs, recent examples of bioactive compounds obtained in the author’slaboratory in projects directed towards the search for novel anticancer agents and cancer chemopre-ventives from higher plants will be presented.In the United States in 1999, it is estimated that over 1.2 million persons will be diagnosed with in-vasive forms of cancer, and over 1,500 people will die as a result of cancer each day [5]. Among manyrecent advances in cancer chemotherapy, plant natural products have played an important role in con-tributing to the arsenal of the approximately 60 cancer chemotherapeutic drugs on the market. For in-stance, in the United States, there are now four structural classes of plant anticancer agents available,constituted by the Catharanthus (Vinca) alkaloids (vinblastine, vincristine, vinorelbine), the epipodo-phyllotoxins (etoposide, etoposide phosphate, teniposide), the taxanes (paclitaxel and docetaxel), andthe camptothecin derivatives (irinotecan and topotecan) [6]. Several other plant-derived compoundsare currently in preclinical and clinical trials [6,7].As part of a National Cooperative Natural Products Drug Discovery Group (NCNPDDG) researchproject funded by the United States National Cancer Institute (1995-2000), our collaborative team atthe College of Pharmacy, University of Illinois at Chicago (Chicago, Illinois), and Research TriangleInstitute (Research Triangle Park, North Carolina), and Bristol-Myers Squibb (Princeton, New Jersey)is evaluating about 400 plant samples per year, with the aim of discovering and evaluating novel plant-derived anticancer agents. During the funding period 1990-1995, the industrial partner was GlaxoWellcome Medicines Research Centre (Sevenage, U.K.), and past progress made in the project hasbeen reviewed [8]. Since 1995, the primary plant samples have been collected in the Dominican Re-public, Peru, and Indonesia. Plant recollections have taken place mainly in Thailand and Zimbabwe in

Two-photon autofluorescence microscopy and spectroscopy of Antarctic fungus: new approach for studying effects of UV-B irradiation.

We combined two-photon fluorescence microscopy and spectroscopy to provide functional images of UV-B (280-315 nm) induced stress on an Antarctic fungus. Two-photon excitation microscopy was used to characterize the distribution of autofluorescence inside the spore and the hyphae of the fungus. The imaging analysis clearly shows that the autofluorescence response of spores is higher than that of hyphae. The imaging analysis at different depths shows that, strikingly enough, the spore autofluorescence originates from the cell wall and membrane fluorophores. The spectroscopic results show moreover that the fluorescence spectra of spores are redshifted upon UV-B irradiation. Tentative identification of the chromophores involved in the autofluorescence response and their biological relevance are also discussed on the basis of a previous steady-state fluorescence spectroscopic study performed on both whole spore suspension and organic-soluble extracts.

Ulosantoin, a potent insecticide from the sponge Ulosa ruetzleri

Ulosantoin (2), a phosphorylated hydantoin, has been isolated from organic soluble extracts of the sponge Ulosa ruetzleri and found to exhibit marked insecticidal activity against tobacco hornworm larvae and cockroaches. The structure was determined by X-ray diffraction analysis. A structurally related compound, dimethyl N 2 -creatininylphosphate (1), was also isolated from the sponge extracts, but was inactive in the insecticidal screens

Extraction Efficiencies for Pesticides in Crops: 14C-Benomyl Extraction from Mustard Greens and Radishes

14C-labeled benomyl [methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate] suspended in a commercial benomyl formulation was sprayed on mustard greens and radishes. At 3 intervals after application, the crops were extracted with methanol, acetonitrile, or acetone. Crops were either blended and leached or repetitively blended followed by Soxhlet extraction. Essentially all of the extractable radioactivity was removed by blending. The 14C was more difficult to extract from radishes than from mustard greens as time increased. Respective percentages of 14C extracted at 1, 7, and 14 days were 99, 98, and 97 from mustard greens and 96, 88, and 79 from radishes. Methanol exhibited the highest extraction efficiency, and the blend-Soxhlet process was better than the blend-leach process. Thin layer chromatography of the organic-soluble extracts indicated that the majority of 14C was recovered as methyl 2-benzimidazole carbamate (MBC), a breakdown product of benomyl. Acid hydrolysis of the extracted tissues released 30--50% of the residual 14C.