Chemical Composition and Cytotoxic Activity of the Essential Oil and Oleoresins of In Vitro Micropropagated Ansellia africana Lindl: A Vulnerable Medicinal Orchid of Africa.

Orchids are rich treasure troves of various important phytomolecules. Among the various medicinal orchids, Ansellia africana stands out prominently in the preparing of various herbal medicines due to its high therapeutic importance. The nodal explants of A. africana were sampled from asymbiotically germinated seedlings on basal Murashige and Skoog (MS) medium and were micropropagated in MS medium supplemented with 3% sucrose and 10 µM meta topolin (mT) + 5 µM naphthalene acetic acid (NAA) +15 µM indole butyric acid (IBA) + 30 µM phloroglucinol (PG). In the present study, the essential oil was extracted by hydrodistillation and the oleoresins by the solvent extraction method from the micropropagated A. africana. The essential oil and the oleoresins were analysed by Gas Chromatography (GC) and GC/MS (Mass spectrometry). A total of 84 compounds were identified. The most predominant components among them were linoleic acid (18.42%), l-ascorbyl 2,6-dipalmitate (11.50%), linolenic acid (10.98%) and p-cresol (9.99%) in the essential oil; and eicosane (26.34%), n-butyl acetate (21.13%), heptadecane (16.48%) and 2-pentanone, 4-hydroxy-4-methyl (11.13%) were detected in the acetone extract; heptadecane (9.40%), heneicosane (9.45%), eicosane (6.40%), n-butyl acetate (14.34%) and styrene (22.20%) were identified and quantified in the ethyl acetate extract. The cytotoxic activity of essential oil and oleoresins of micropropagated A. africana was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide) assay on Vero cells compared to the standard drug doxorubicin chloride. The present research contains primary information about the therapeutic utility of the essential oil and oleoresins of A. africana with a promising future research potential of qualitative and quantitative improvement through synchronised use of biotechnological techniques.

Promoting the treatment of crude oil alkane pollution through the study of enzyme activity.

Microbes appear to play a key role in bioremediation of petroleum hydrocarbons pollution and little attention has been paid to the enzyme activity in the process of alkane bioremediation. Oil field bacterium identified as Pseudomonas synxantha LSH-7' was chosen as the tested strain. Periodically collected samples were analyzed by GC-FID (Gas Chromatography- Flame Ionization Detector) and RT-qPCR (Quantitative-Real-Time-PCR). GC-FID results showed this bacterial strain has great degradation ability on crude oil n-alkanes and RT-qPCR data indicated the differences between the three genes expression including AlkB-, Cytochromes P450-, and almA- related when grown on different-chain alkanes. Meanwhile, enzyme activity like alkane hydroxylase, alcohol dehydrogenase, dehydrogenase, protease, phosphatase, catalase and lipase were measured. Extracellular alkane hydroxylase was induced in a higher degree than intracellular in the early incubation time, alcohol dehydrogenase increased/decreased along with alkane hydroxylase, and the pH of the medium obviously decreased. Other enzymes were also described including dehydrogenase activity that reached a highest point that was slower than alcohol dehydrogenase, protease activity started multiplying after a period of culture while biomass was immediately increased, catalase activity dramatically enhanced in the presence of alkanes, phosphatase activity was closely linked to pH approximately but lipase activity was found to be moderate.

Bioactive compounds in Pinang Yaki (Areca vestiaria) fruit as potential source of antifertility agent.

Fruits of Pinang Yaki (Areca vestiaria) are used by the people around Bogani Nani Wartabone as contraseption for men. Extracts from the fruit contain tannin, triterpenoid, flavonoid and saponin which are potential as bioactive compounds. This research aimed at exploring the fractions or bioactive compounds contained in the fruit. The extract was prepared by fractionation using hexane. The fractions were separated and analysed by gas chromatography mass spectrometry (GC-MS) technique. The fractions revealed the presence of five compounds. These compounds were identified by interpretation of mass spectra and comparing their retention time and covate indexes with those from literature. The five compounds are pentadecane, methyl-dodecanate, methyl-tetradecanoate, hexadecanoic acid and methyl-octadecanate.

A new flavanolignan and a new alkane from the Stem bark of Newtonia griffoniana.

Two new compounds a flavanolignan (1), and an alkane (2) along with four known compounds including two fatty acid esters (3-4) and two isocoumarins (5-6) were isolated from the methanolic extract of the stem bark of Newtonia griffoniana. Their structures were elucidated using spectroscopic methods including extensive 1-D and 2-D NMR experiments.

Bridging the gap between gas and liquid chromatography.

The rapid and complete baseline separation of both volatile (C5 to C16 alkanes in gasoline or terpenes in plant extracts) and non-volatile (>C20 alkanes) organic compounds was achieved by combining (1) low-density fluid chromatography (LDFC) using carbon dioxide at elevated temperature (>90°C) and low pressure (1500psi) designed to increase the retention of the most volatile compounds and (2) high-vacuum technology (<10-4Torr) in order to preserve the maximum efficiency of short analytical columns (3.0mm×150mm packed with 1.8µm fully porous HSS-SB-C18 particles) when used in LDFC. The volatile compounds are eluted first under isobaric conditions (1500psi) in less than a minute followed by a linear gradient of the column back pressure (from 1500 to 3500psi in 5min) for the elution of the non-volatile compounds up to C40. The experimental results demonstrate that LDFC performed with short 3.0mm i.d. columns packed with sub-2µm particles and placed under adiabatic conditions enables the analysts to deliver a single, fast, and high-resolution separation of both volatile and non-volatile compounds.

A new furanosteroid from Talaromyces sp. lgt-4, a fungal endophyte isolated from Tripterygium wilfordii.

Wortmannolol (1), a new furanosteroid, along with five known compounds, wortmannolone (2), ergosterol (3), p-hydroxyphenyl ethanol (4), trans-6-dodecene (5), (2Z, 4E) -5-(8-hydroxy-1,5-dimethyl-3-oxo-6-oxabicyclo [3.2.1] octan-8-yl) -3-methylpenta-2,4-dienoic acid (6) were isolated from a fungal endophyte Talaromyces sp. lgt-4. Their structures were elucidated by IR, MS, 1D and 2D NMR spectra. Compound 1 show weak monoamine oxidase inhibitory activity.

Effect of dispersants on the biodegradation of South Louisiana crude oil at 5 and 25 °C.

This article reports biodegradation rates for a commercial dispersant, JD-2000, South Louisiana crude oil (SLC) alone, and SLC dispersed with JD-2000 at 5 and 25 °C. Results from the biodegradation experiments revealed that Component X, a chemical marker for JD-2000, rapidly degraded at both temperatures. The application of JD-2000 decreased by half the overall biodegradation rate of aliphatic compounds at 25 °C. At 5 °C, a residual fraction consisting of iso- and n-alkanes (C29-C35) persisted after 56 d. The combination of dispersant and higher temperature resulted in faster removal rates for 2- and 3-ring polycyclic aromatic hydrocarbons. When compared with Corexit 9500, our results suggest that the chemistry of the surfactant (or surfactants) in JD-2000 might have favored oil dissolution (substrate transport to the aqueous phase) as an uptake mechanism over adhesion, which requires direct contact of the biomass with the oil.

Granular encapsulation of light hydrophobic liquids (LHL) in LHL-salt water systems: Particle induced densification with quartz sand.

Addition of granular materials to floating crude oil slicks can be effective in capturing and densifying the floating hydrophobic phase, which settles by gravity. Interaction of light hydrophobic liquids (LHL) with quartz sand was investigated in LHL-salt water systems. The LHLs studied were decane, tetradecane, hexadecane, benzene, toluene, ethylbenzene, m-xylene, and 2-cholorotoluene. Experiments were conducted with fine quartz sand (passing sieve No. 40 with openings 0.425 mm). Each LHL was dyed with few crystals of Sudan IV dye for ease of visual observation. A volume of 0.5 mL of each LHL was added to 100 mL salt water (34 g/L). Addition of one gram of quartz sand to the floating hydrophobic liquid layer resulted in formation of sand-encapsulated globules, which settled due to increased density. All LHLs (except for a few globules of decane) formed globules covered with fine sand particles that were heavy enough to settle by gravity. The encapsulated globules were stable and retained their shape upon settling. Polarity of hydrophobic liquids as the main factor of aggregation with minerals was found to be insufficient to explain LHL aggregation with sand. Contact angle measurements were made by submerging a large quartz crystal with the LHL drop on its surface into salt water. A positive correlation was observed between the wetting angle of LHL and the LHL volume captured (r = 0.75). The dependence of the globule density on globule radius was analyzed in relation to the coverage (%) of globule surface (LHL-salt water interface) by fine quartz particles.

Free radical scavenging capacity and protective effect of natural substances in peloids from the thermal spring pool Bagnaccio (Viterbo, Italy).

Natural peloids from sulfurous thermal springs are largely used in cosmetic and pelotherapy for the treatment of different dermatological conditions, including skin aging, dermatitis, and other eczemas. The beneficial effects are correlated to mineralogical and other thermal properties, as well as to the presence of natural substances with specific antioxidant activity. Few data are available for the comparison between natural peloids and synthetic (i.e., artificially maturated) muds. In this context, the natural substances and antioxidant activity of natural white mud (WM) and dark mud (DM) peloids from the sulfurous thermal spring pool Bagnaccio (Viterbo, Italy) have been studied in detail to evaluate possible relationships between physicochemical properties and therapeutic effect. A large panel of natural substances in WM and DM were characterized for the first time by ³¹P-nuclear magnetic resonance and gas chromatography associated to mass spectrometry analysis. Polar fractions of WM and DM peloids were characterized by the presence of several bioactive natural compounds, showing high antioxidant activity and DNA protective effect, as evaluated by 2,2-diphenyl-1-picrylhydrazyl assay, and hydrogen peroxide­induced DNA breakage in the alkaline comet assay. The antioxidant activity and DNA protective effect could be attributed to radical scavenging rather than a modulatory effect on the induced DNA repair, and are of order of intensity higher than that reported for synthetic muds.

Characteristic Chemical Components and Aroma-active Compounds of the Essential Oils from Ranunculus nipponicus var. submersus Used in Japanese Traditional Food.

Ranunculus nipponicus var. submersus is an aquatic macrophyte; it is known as a wild edible plant in Japan for a long time. In this study, the essential oils from the fresh and dried aerial parts of R. nipponicus var. submersus were extracted by hydrodistillation and analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). Moreover, important aroma-active compounds were also detected in the oil using GC-olfactometry (GC-O) and aroma extract dilution analysis (AEDA). Thus, 98 compounds (accounting for 93.86%) of the oil were identified. The major compounds in fresh plant oil were phytol (41.94%), heptadecane (5.92%), and geranyl propionate (5.76%), while those of. Dried plant oil were ß-ionone (23.54%), 2-hexenal (8.75%), and dihydrobovolide (4.81%). The fresh and dried oils had the green-floral and citrus-floral odor, respectively. The GC-O and AEDA results show that phenylacetaldehyde (green, floral odor, FD-factor = 8) and ß-ionone (violet-floral odor, FD-factor = 8) were the most characteristic odor compounds of the fresh oils. ß-Cyclocitral (citrus odor, FD-factor = 64) and ß-ionone (violet-floral odor, FD-factor = 64) were the most characteristic odor compounds of the dried oil. These compounds are thought to contribute to the flavor of R. nipponicus var. submersus.

Chemical composition of the essential oil of Jacobaea maritima (L.) Pelser & Meijden and Jacobaea maritima subsp. bicolor (Willd.) B. Nord. & Greuter (Asteraceae) collected wild in Croatia and Sicily, respectively.

In this study, the chemical compositions of the essential oils from aerial parts (JmA) and radices (JmR) of Jacobaea maritima (L.) Pelser & Meijden, collected in Croatia, and of Jacobaea maritima subsp. bicolor (Willd.) B. Nord. & Greuter, collected in Sicily, were evaluated by using gas chromatography and gas chromatography-mass spectrometry. The main components of the essential oil from J. maritima, both in JmA and JmR, were pentacosane (15.7%), heptacosane (13.1%) and nonacosane (8.1%) whereas the essential oil from J. maritima subsp. bicolor was characterised by the presence of hexadecanoic acid (14.6%), caryophyllene oxide (9.3%) and hexahydrofarnesylacetone (6.5%). The comparison of the essential oil with other studied oils of the genus Jacobaea is discussed.

Acaricidal activity of petroleum ether extract of leaves of Tetrastigma leucostaphylum (Dennst.) Alston against Rhipicephalus (Boophilus) annulatus.

The acaricidal activity of the petroleum ether extract of leaves of Tetrastigma leucostaphylum (Dennst.) Alston (family: Vitaceae) against Rhipicephalus (Boophilus) annulatus was assessed using adult immersion test (AIT). The per cent of adult mortality, inhibition of fecundity, and blocking of hatching of eggs were studied at different concentrations. The extract at 10% concentration showed 88.96% inhibition of fecundity, 58.32% of adult tick mortality, and 50% inhibition of hatching. Peak mortality rate was observed after day 5 of treatment. Mortality of engorged female ticks, inhibition of fecundity, and hatching of eggs were concentration dependent. The LC50 value of the extract against R. (B.) annulatus was 10.46%. The HPTLC profiling of the petroleum ether extract revealed the presence of at least seven polyvalent components. In the petroleum ether extract, nicotine was identified as one of the components up to a concentration of 5.4%. However, nicotine did not reveal any acaricidal activity up to 20000 ppm (2%). Coconut oil, used as diluent for dissolving the extract, did not reveal any acaricidal effects. The results are indicative of the involvement of synergistic or additive action of the bioactive components in the tick mortality and inhibition of the oviposition.

Sorption and distribution of asphaltene, resin, aromatic and saturate fractions of heavy crude oil on quartz surface: molecular dynamic simulation.

The molecular scale sorption, diffusion and distribution of asphaltene, resin, aromatic and saturate fractions of heavy crude oil on quartz surface were studied using molecular dynamic simulation. Sorption of saturates on quartz decreased by 31% when temperature increased from 298 to 398K while opposite trend was observed for resins, but insignificant changes were found in asphaltenes and aromatics. Despite of this variety, the main contribution of interactions was van der Waals energy (>90%) irrespective of molecular components and temperatures. The diffusion coefficient of saturates was predicted as 10.8×10(-10)m(2)s(-1) while that of the remaining fractions was about 4×10(-10)m(2)s(-1) at 298K. The most likely oil distribution on quartz surface was that aromatics and saturates transported randomly into and out of the complex consisting of asphaltenes surrounded by resins, which was influenced by temperature. Overall, the knowledge on quartz-oil and oil-oil interactions gained in this study is essential for future risk assessment and remediation activities as previous studies on soil remediation either limited to light oil fractions with <40 carbons or treated the heavy crude oil as a single pseudo entity ignoring the interactions between oil fractions.

[Study on chemical constituents from Anoectochilus chapaensis].

OBJECTIVE: To investigate the chemical constituents from the whole herbs of Anoectochilus chapaensis. METHODS: The chemical constituents were isolated and purified by repeated column chromatographies with silica gel, Sephadex LH-20 and preparative TLC. Their structures were identified by their physiochemical property and spectral data. RESULTS: 6 compounds were isolated and elucidated as follows: (1) Friedelin, (2) Sorghumol, (3) 5alpha, 8alpha-epidioxyergost-22-en-3beta-ol, (4) Stearic acid, (5) Octadecane and (6) Epifriedelanol. CONCLUSION: Compound (1), (2) and (4) are isolated from this plant for the first time, and compound (3), (5) and (6) are isolated from genera Anoectochilus for the first time.

[Analysis of petroleum fraction from flower, stem and leaf of Aconitum taipeicum by GC-MS].

OBJECTIVE: To analyze the chemical constituents of petroleum fraction of Aconitum taipeicum. METHODS: The methanol extract of Aconitum taipeicum were extracted by petroleum and then analyzed by GC-MS. The compounds were quantitatively determined by normalization method. RESULTS: Twenty eight compounds were separated and identified. Most of them were alkane, fat acids and their esters and alkenes. The Nonacosane covered 13.057% of the total peaks, while 19-methyl-18,21-Hexatriacontanediether 8.180%, Ethylen eglycol monooctadecy ether 7.851%, 3,7,11,15-Tetramethyl-2-hexadecen-1-ol 7.805%, Metahyl Palmitate 6.676% and so on. CONCLUSION: This is the first report of constituents from the flower, stem and leaf of Aconitum taipeicum. The results will provide foundation for further exploitation and use of Aconitum taipeicum.

Evaluation of cytotoxic activity of patriscabratine, tetracosane and various flavonoids isolated from the Bangladeshi medicinal plant Acrostichum aureum.

CONTEXT: Acrostichum aureumL. (Pteridaceae), a mangrove fern, has been used as a Bangladeshi traditional medicine for a variety of diseases including peptic ulcer. OBJECTIVE: Isolation and structural elucidation of cytotoxic secondary metabolites from the methanol extract of the aerial parts of A. aureum. MATERIALS AND METHODS: Compounds were isolated using HPLC. The compound structures were elucidated by 1D and 2D NMR, MS and other spectroscopic methods using published data. The compounds were tested for their cytotoxic activity against healthy and cancer cells using the MTT assay. Active compounds were further evaluated for apoptosis-and necrosis-inducing potential against gastric cancer cells (AGS) using the FITC Annexin V apoptosis assay. RESULTS AND DISCUSSION: Seven known compounds, patriscabratine, tetracosane and 5 flavonoids (quercetin-3-O-ß-d-glucoside, quercetin-3-O-ß-d-glucosyl-(6→1)-α-l-rhamnoside, quercetin-3-O-α-l-rhamnoside, quercetin-3-O-α-l-rhamnosyl-7-O-ß-d-glucoside and kaempferol) were isolated. Patriscabratine was found moderately cytotoxic against AGS, MDA-MB-231 and MCF-7 cells with IC(50) values ranging from 69.8 to 197.3 µM. Tetracosane showed some cytotoxic activity against AGS, MDA-MB-231, HT-29 and NIH 3T3 cells with IC(50) values ranging from 128.7 to >250 µM. Patriscabratine and tetracosane displayed an apoptotic effect (10%) on AGS cells within 24 h which was increased (20%) after 48 h, and was comparable to, if not greater, than the positive control, cycloheximide. CONCLUSION: Except for quercetin-3-O-ß-d-glucoside and kaempferol; compounds were isolated for the first time from this plant and evaluated for their cytotoxic activity. The results highlight the potential of this plant as a source of bioactive compounds and provide a rationale for its traditional use in peptic ulcer treatment.

Lipophilic extractives from the cortex and pith of elephant grass ( Pennisetum purpureum Schumach.) stems.

The composition of lipophilic extractives in the cortex and pith of elephant grass ( Pennisetum purpureum Schumach.) stems was thoroughly studied by gas chromatography-mass spectrometry. The predominant compounds were fatty acids followed by sterols (in free and conjugated forms as esters and glycosides). Other steroid compounds, as steroid hydrocarbons and ketones, were also present. Additionally, important amounts of mono-, di-, and triglycerides were identified. Other aliphatic series such as n-alkanes, n-fatty alcohols, and n-alkyl ferulates, together with tocopherols and a series of high molecular weight esters, were also found, although in minor amounts. The analyses also revealed the presence of a ß-diketone (12,14-tritriacontanedione), which was particularly abundant in the cortex. Finally, two lignans, matairesinol and syringaresinol, were also detected. In general terms, the abundances of the different classes of compounds were higher in the pith, except for the series of n-fatty alcohols, n-alkyl ferulates, ß-diketones, and lignans, which were more prominent in the cortex.

Production of isotopically labeled standards from a uniformly labeled precursor for quantitative volatile metabolomic studies.

Optimal accuracy and precision in small-molecule profiling by mass spectrometry generally requires isotopically labeled standards chemically representative of all compounds of interest. However, preparation of mixed standards from commercially available pure compounds is often prohibitively expensive and time-consuming, and many labeled compounds are not available in pure form. We used a single-prototype uniformly labeled [U-(13)C]compound to generate [U-(13)C]-labeled volatile standards for use in subsequent experimental profiling studies. [U-(13)C]-α-Linolenic acid (18:3n-3, ALA) was thermally oxidized to produce labeled lipid degradation volatiles which were subsequently characterized qualitatively and quantitatively. Twenty-five [U-(13)C]-labeled volatiles were identified by headspace solid-phase microextraction-gas chromatography/time-of-flight mass spectrometry (HS-SPME-GC/TOF-MS) by comparison of spectra with unlabeled volatiles. Labeled volatiles were quantified by a reverse isotope dilution procedure. Using the [U-(13)C]-labeled standards, limits of detection comparable to or better than those of previous HS-SPME reports were achieved, 0.010-1.04 ng/g. The performance of the [U-(13)C]-labeled volatile standards was evaluated using a commodity soybean oil (CSO) oxidized at 60 °C from 0 to 15 d. Relative responses of n-decane, an unlabeled internal standard otherwise absent from the mixture, and [U-(13)C]-labeled oxidation products changed by up to 8-fold as the CSO matrix was oxidized, demonstrating that reliance on a single standard in volatile profiling studies yields inaccurate results due to changing matrix effects. The [U-(13)C]-labeled standard mixture was used to quantify 25 volatiles in oxidized CSO and low-ALA soybean oil with an average relative standard deviation of 8.5%. Extension of this approach to other labeled substrates, e.g., [U-(13)C]-labeled sugars and amino acids, for profiling studies should be feasible and can dramatically improve quantitative results compared to use of a single standard.

The detection of high-mass aliphatics in petroleum by matrix-assisted laser desorption/ionisation mass spectrometry.

RATIONALE: The development of simplified procedures for isolating high-mass alkanes present in crude oils is described. The new procedures, which bypass the sample recovery step with hot toluene in the conventional alkane-isolation procedure, also provide an effective sample preparation route, prior to analysis by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS). METHODS: Urea-alkane adducts are formed by mixing sample and urea solutions on chromatographic paper or silica-coated plates. Unreacted hydrocarbons are removed by developing the plates with chloroform. In a second development with water, adducts are broken up in situ and the liberated urea removed, leaving bands of isolated alkanes behind. For MALDI-MS, strips of paper/plates, carrying the isolated alkanes, are fixed on metal target plates. The samples are treated with matrix (AgNO(3)) and analysed by MALDI-MS. RESULTS: The observed signal represents silver ion adducts of the isolated alkanes. Silver appears to work without much fragmentation and to generate whole silver adduct ions. Much improved MALDI-MS detection sensitivity and a wider range of masses was observed when samples were ablated from paper/plate surfaces, than by ablation from bulk samples spread over a smooth surface--the conventional method. Chromatographic paper gave better resolution and a broader range of masses than silica-coated plates. CONCLUSIONS: The analytical sequences have been confirmed using standard alkanes (C(20)-C(60)) and Polywax. The proposed procedures enhanced the sensitivity and detection range of the MS analysis. The method was useful in detecting n-alkanes to m/z 1500 (C(100)) and required relatively small quantities of sample and reagents. It provides a promising qualitative analysis route for the rapid isolation and reliable determination of alkanes in crude oils.