Chemical variability of the wood essential oil of Cedrus atlantica Manetti from Corsica

The composition of 48 samples of essential oil isolated from the wood of Cedrus atlantica growing in Corsica was investigated by GC (in combination with retention indices), GC/MS, and (13) C-NMR. Twenty-three compounds accounting for 73.9-96.0% of the oil composition were identified. The oils consisted mainly of monoterpene hydrocarbons and sesquiterpenes, in particular α-pinene (5; up to 79.4%), himachalol (4; up to 66.2%), β-pinene (up to 21.4%), β-himachalene (2; up to 19.3%), γ-himachalene (3; up to 11.0%), and α-himachalene (1; up to 10.9%). The 48 oil compositions were submitted to k-means partitioning and principal-component analysis, which allowed the distinction of two groups within the oil samples. The composition of Group I (44% of the samples) was dominated by 5, while the samples of Group II (56% of the samples) contained mainly 4.     

Isolation, Structure elucidation, and antimycobacterial properties of dimeric naphtho-gamma-pyrones from the marine-derived fungus Aspergillus carbonarius

Two new dimeric naphtho-gamma-pyrones, compounds 1 and 2, were isolated from the AcOEt extract of the fungal strain WZ-4-11 of Aspergillus carbonarius, together with eight known analogues, including 10,10'-bifonsecin B (3), 6'-O-demethylnigerone (4), nigerone (5), isonigerone (6), fonsecin (7), rubrofusarin B (8), TMC 256A1 (9), and flavasperone (10). Their structures were elucidated by means of UV, CD, IR, and 1D- and 2D-NMR spectroscopy, in combination with HR-MS analysis. The fully assigned (1)H- and (13)C-NMR data of 3, and the (13)C-NMR data of 6 are reported for the first time. Compounds 1 and 2 showed weak antimycobacterial activities against Mycobacterium tuberculosis H37Rv, with MIC values of 43.0 and 21.5 microM, resp.     

Composition of the leaf, stem bark and root bark oils of Isolona cooperi investigated by GC (retention index), GC-MS and 13C-NMR spectroscopy

The leaf, stem bark and root bark oils of Isolona cooperi Hutchinson & Dalziel from the Ivory Coast have been analysed by GC (retention index), GC-MS and 13C-NMR spectroscopy. Two types of essential oil were produced by the plant. The leaf and stem bark oils were monoterpene-rich, containing principally (Z)-beta-ocimene and gamma-terpinene and three lactones, 5-[(E and Z)-hexylidene]-5H-furan-2-ones and massoia lactone, were present in appreciable amounts. Conversely, the root bark oil was dominated by 5-isopentenylindole and (E)-beta-caryophyllene. The strategy for the analysis of each oil was adapted according to the nature of the components.     

Composition and Chemical Variability of the Essential Oil from Aerial Parts of Cladanthus scariosus,an Endemic Species to the Moroccan High Atlas

Cladanthus scariosus (Ball) Oberpr. & Vogt is endemic to Moroccan High Atlas. It is known under the vernacular names Irezghi or Irezgui. Three essential oil samples have been isolated from aerial parts and analyzed by combination of chromatographic and spectroscopic techniques [gas chromatography (GC) in combination with retention indices (RI), gas chromatography-mass spectrometry (GC/MS) and ¹³C-NMR spectroscopy]. The compositions of oil samples were dominated by monoterpenes: α-pinene sabinene, and terpinen-4-ol. Chamazulene and dihydrochamazulene isomers as well as various hemiterpene esters and analogs have been identified. To evidence a chemical variability, statistical analysis performed on 13 oil sample compositions allowed partitioning into three groups, mainly differentiated by their contents of sabinene, camphor, borneol, terpinen-4-ol, and germacrene D.     

Separation and purification of four chromones from radix saposhnikoviae by high-speed counter-current chromatography

A preparative high-speed counter-current chromatography (HSCCC) method for the isolation and purification of 1'-O-glucosylcimifugin (1), 4'-O-beta-d-glucosyl-5-O-methylvisamminol (2), cimifugin (3) and 3'-O-glucosylhamaudol (4) from the Chinese medicinal herb radix saposhnikoviae has been successfully developed. A sample of 300 mg of crude extract was separated using ethyl acetate:n-butanol:1% aqueous acetic acid (1:4:5, v/v) as the two-phase solvent system and yielded 102.4 mg of 1 and 81.6 mg of 2. During this separation 3 and 4 remained in the stationary phase, which was collected, evaporated to dryness and separated with another two-phase solvent system involving ethyl acetate:n-butanol:1% aqueous acetic acid (5:0.5:5, v/v) to yield 31.4 mg of 3 and 12.7 mg of 4. The purities of compounds 1-4 were 98.4, 98.7, 99.3 and 98.2%, respectively, as determined by HPLC. The chemical structures of these components were established by (1)H-NMR and (13)C-NMR.     

Analysis of the essential oil from the roots of Eupatorium cannabinum subsp. corsicum (L.) by GC, GC-MS and 13C-NMR

Eupatorium cannabinum subsp. corsicum (L.) is an endemic subspecies from the island of Corsica. The essential oil from the roots of this aromatic plant has been studied by GC, GC-MS and by 13C-NMR. In contrast to the essential oil from the aerial parts, which is dominated by hydrocarbon compounds (76.9%) and particularly by sesquiterpene components (43.3%), the essential oil from the roots was characterized by a high content of oxygenated compounds (61.0%), particularly oxygenated monoterpenes (54.0%). In the root oil, 106 components were identified representing 96.1% of the total amount. This oil was dominated by the monoterpenes esters (33%), the major components of which were neryl isobutyrate (17.6%), thymyl methyl oxide (15.1%), delta-2-carene (14.5%) and beta-pinene (5.7%). Aromatic esters, nerol derivatives (esters and diesters) and a benzofuran were investigated by GC-MS using different ionization modes including electron impact ionization, and positive- and negative-chemical ionization. These components have not previously been reported in the essential oil of aerial parts of E. cannabinum from Corsica island.     

Studies on Triterpenoid Constituents Isolated from the Roots of Sabia schumanniana

Five triterpenoids were isolated from the roots of Sabia schurnanniana Diels. Based on their physical constants, UV. IR, 1H-NMR, 13C-NMR, DEPT spectroscopic analysis and preparation of their derivatives, they were identified as 3-oxo-olean-ll, 13 (18)-diene (1), 3, 1 1-dioxo-olean- 12-ene (2), 3β -hydroxy-olean- 11, 13 (1 8) -diene (3), 3-oxo, 11 α-hydroxy-olean-12-ene (4), 3, 11α-dihydroxy-olean-12-ene (5) . Compound 4 is a new compound. Their 13C-NMR spectra have been signified by means of DEPT analysis.     

The Volatile Profiles of a Rare Apple (Malus domestica Borkh.) Honey: Shikimic Acid‐Pathway Derivatives,Terpenes, and Others

The volatile profiles of rare Malus domestica Borkh . honey were investigated for the first time. Two representative samples from Poland (sample I) and Spain (sample II) were selected by pollen analysis (44–45% of Malus spp. pollen) and investigated by GC/FID/MS after headspace solid‐phase microextraction (HS‐SPME) and ultrasonic solvent extraction (USE). The apple honey is characterized by high percentage of shikimic acid‐pathway derivatives, as well as terpenes, norisoprenoids, and some other compounds such as coumaran and methyl 1H‐indole‐3‐acetate. The main compounds of the honey headspace were (sample I; sample II): benzaldehyde (9.4%; 32.1%), benzyl alcohol (0.3%; 14.4%), hotrienol (26.0%, 6.2%), and lilac aldehyde isomers (26.3%; 1.7%), but only Spanish sample contained car‐2‐en‐4‐one (10.2%). CH₂Cl₂ and pentane/Et₂O 1 : 2 (v/v) were used for USE. The most relevant compounds identified in the extracts were: benzaldehyde (0.9–3.9%), benzoic acid (2.0–11.2%), terpendiol I (0.3–7.4%), coumaran (0.0–2.8%), 2‐phenylacetic acid (2.0–26.4%), methyl syringate (3.9–13.1%), vomifoliol (5.0–31.8%), and methyl 1H‐indole‐3‐acetate (1.9–10.2%). Apple honey contained also benzyl alcohol, 2‐phenylethanol, (E)‐cinnamaldehyde, (E)‐cinnamyl alcohol, eugenol, vanillin, and linalool that have been found previously in apple flowers, thus disclosing similarity of both volatile profiles.     

Larvicidal and Nematicidal Activities of the Leaf Essential Oil of Croton regelianus

The chemical composition of the leaf essential oil of Croton regelianus collected from wild plants growing in two different sites at Ceará State (Brazil) was analyzed by GC/MS and GC‐FID. Twenty monoterpenoids, representing more than 96% of the chemical composition of the oils, were identified and quantified. The oils showed similar chemical composition but considerable variation in the levels of each constituent. Ascaridole (33.9–17.0%), p‐cymene (22.3–21.6%), and camphor (13.0–3.1%) were the predominant constituents. The monoterpene ascaridole was isolated and characterized by spectroscopic data. The essential oils and the isolated compounds were tested against Aedes aegypti and Artemia sp. larvae, and the root knot nematode Meloidogyne incognita. The bioassay results show that the essential oil of C. regelianus and ascaridole were moderately active against the M. incognita, but strongly effective against both A. aegypti and Artemia sp. larvae.     

Evaluation of Daphne genkwa diterpenes: fingerprint and quantitative analysis by high performance liquid chromatography

Daphne genkwa contains a novel class of anticancer diterpene esters that inhibit DNA topoisomerase I. Fingerprint and quantitative analysis by HPLC were performed in order to characterise and evaluate D. genkwa. A standard fingerprint of Daphne diterpene esters from the root extract was first established by HPLC-UV, and the major peaks in the fingerprint profile were preliminarily determined using HPLC-MS. The principal Daphne diterpene esters, yuanhuacine (1), yuanhuadine (2), yuanhuajine (3) and yuanhuagine (4), were isolated and identified using a combination of UV, IR, MS, 1H-NMR and 13C-NMR spectral data. Quantitative analysis indicated that 1 was the principal component in the root, and that 2 was the major component in the buds. The average extraction rates of 1 and 2 were 0.0151 and 0.0033% (n=10) from the root, respectively, and 0.0020 and 0.0078% (n=3) from the buds, respectively.     

Buzonamine, a new alkaloid from the defensive secretion of the millipede, Buzonium crassipes

GC-MS analysis of the defensive secretion from the millipede Buzonium crassipes showed three volatile components, beta-pinene (35%), limonene (6%) and a new alkaloid, buzonamine (59%). Buzonamine had an HRMS molecular ion at m/z=221.1785 (calculated for C(14)H(23)NO, 221.1781), 14 carbons in the 13C-NMR and 23 hydrogens by SFORD, DEPT and APT experiments. All distinct 1H nuclei were assigned to 13C resonances with 2D 1H-13C COSY data, and the final structure was determined by 1H-1H COSY, 1H-13C HMBC and nOe experiments. Buzonamine contains four rings including an epoxy group and a tertiary nitrogen. beta-Pinene, limonene or buzonamine, kept the mound nesting ant, Formica obscuripes, from eating mealworms (Tenebrio molitor) treated with 1mg of the chemical. A 150mg millipede expels 4mg of secretion.     

Biosynthesis of ascaridole: iodide peroxidase-catalyzed synthesis of a monoterpene endoperoxide in soluble extracts of Chenopodium ambrosioides fruit

Ascaridole, an asymmetric monoterpene endoperoxide with anthelmintic properties, occurs as a major constituent (60-80%) in the volatile oil of American wormseed fruit (Chenopodium ambrosioides: Chenopodiaceae), and as a lesser component in the leaf pocket oil of the boldo tree (Peumus boldus: Monimiaceae). Determination of optical activity and chromatographic resolution of naturally occurring ascaridole, and several synthetic derivatives, showed that both wormseed and boldo produce ascaridole in racemic form. The biosynthesis of ascaridole from the conjugated, symmetrical diene alpha-terpinene (a major component of the oil from wormseed) was shown to be catalyzed by a soluble iodide peroxidase isolated from homogenates of C. ambrosioides fruit and leaves. The enzymatic synthesis of ascaridole was confirmed by capillary gas-liquid chromatography and mass spectrometry of the product, which was also shown to be racemic. Optimal enzymatic activity occurred at pH 4.0 in the presence of 2.5 mM H2O2 and 1 mM NaI. Soluble enzyme extracts were fractionated by gel filtration on both Sephacryl S-300 and Sephadex G-100, and were shown to consist of a high-molecular-weight peroxidase component (Mr greater than 1,000,000, 30% of total activity) and two other peroxidase species having apparent molecular weights of 62,000 and 45,000 (major component). Peroxidase activity was susceptible to proteolytic destruction only after periodate treatment, suggesting an association of the enzyme(s) with polysaccharide material. Ascaridole biosynthesis from alpha-terpinene was inhibited by cyanide, catalase, and reducing agents, but not by compounds that trap superoxide or quench singlet oxygen. A peroxide transfer reaction initiated by peroxidase-generated I+ is proposed for the conversion of alpha-terpinene to ascaridole.     

碱提水溶性安络小皮伞多糖的研究——B_1 与 B_2 的分离纯化及结构研究

从曾用水提取过水溶性多糖的安络小皮伞残渣中用稀碱液提取了二种水溶多糖 B_1和B_2。它们分别给 Sepharose 4B 柱,2B 柱层析,超离心分析,醋酸纤维薄膜电泳鉴定,表明 B_1和 B_2均为纯多糖。采用气相色谱,红外光谱,淀粉酶解,甲基化分析等方法确定了它的基本结构。     

Chemometric Profile of Root Extracts of Rhodiola imbricata Edgew. with Hyphenated Gas Chromatography Mass Spectrometric Technique

Rhodiola imbricata Edgew. (Rose root or Arctic root or Golden root or Shrolo), belonging to the family Crassulaceae, is an important food crop and medicinal plant in the Indian trans-Himalayan cold desert. Chemometric profile of the n-hexane, chloroform, dichloroethane, ethyl acetate, methanol, and 60% ethanol root extracts of R. imbricata were performed by hyphenated gas chromatography mass spectrometry (GC/MS) technique. GC/MS analysis was carried out using Thermo Finnigan PolarisQ Ion Trap GC/MS MS system comprising of an AS2000 liquid autosampler. Interpretation on mass spectrum of GC/MS was done using the NIST/EPA/NIH Mass Spectral Database, with NIST MS search program v.2.0g. Chemometric profile of root extracts revealed the presence of 63 phyto-chemotypes, among them, 1-pentacosanol; stigmast-5-en-3-ol, (3β,24S); 1-teracosanol; 1-henteracontanol; 17-pentatriacontene; 13-tetradecen-1-ol acetate; methyl tri-butyl ammonium chloride; bis(2-ethylhexyl) phthalate; 7,8-dimethylbenzocyclooctene; ethyl linoleate; 3-methoxy-5-methylphenol; hexadecanoic acid; camphor; 1,3-dimethoxybenzene; thujone; 1,3-benzenediol, 5-pentadecyl; benzenemethanol, 3-hydroxy, 5-methoxy; cholest-4-ene-3,6-dione; dodecanoic acid, 3-hydroxy; octadecane, 1-chloro; ethanone, 1-(4-hydroxyphenyl); α-tocopherol; ascaridole; campesterol; 1-dotriacontane; heptadecane, 9-hexyl were found to be present in major amount. Eventually, in the present study we have found phytosterols, terpenoids, fatty acids, fatty acid esters, alkyl halides, phenols, alcohols, ethers, alkanes, and alkenes as the major group of phyto-chemotypes in the different root extracts of R. imbricata. All these compounds identified by GC/MS analysis were further investigated for their biological activities and it was found that they possess a diverse range of positive pharmacological actions. In future, isolation of individual phyto-chemotypes and subjecting them to biological activity will definitely prove fruitful results in designing a novel drug.     

Variability of cork from Portuguese Quercus suber studied by solid-state (13)C-NMR and FTIR spectroscopies.

A new approach is presented for the study of the variability of Portuguese reproduction cork using solid-state (13)C-NMR spectroscopy and photoacoustic (PAS) FTIR (FTIR-PAS) spectroscopy combined with chemometrics. Cork samples were collected from 12 different geographical sites, and their (13)C-cross-polarization with magic angle spinning (CP/MAS) and FTIR spectra were registered. A large spectral variability among the cork samples was detected by principal component analysis and found to relate to the suberin and carbohydrate contents. This variability was independent of the sample geographical origin but significantly dependent on the cork quality, thus enabling the distinction of cork samples according to the latter property. The suberin content of the cork samples was predicted using multivariate regression models based on the (13)C-NMR and FTIR spectra of the samples as reported previously. Finally, the relationship between the variability of the (13)C-CP/MAS spectra with that of the FTIR-PAS spectra was studied by outer product analysis. This type of multivariate analysis enabled a clear correlation to be established between the peaks assigned to suberin and carbohydrate in the FTIR spectrum and those appearing in the (13)C-CP/MAS spectra.     

Novel DPPH radical scavengers, demethylbisorbibutenolide and trichopyrone, from a fungus

In our screening program for antioxidants with 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity, two novel compounds, demethylbisorbibutenolide (1) and trichopyrone (2), were isolated from the fermentation broth of the fungus of USF-4860 strain isolated from a soil sample. The structures of these compounds were determined from spectroscopic evidence. The biosynthetic origin of the carbon atoms of 2 was unambiguously determined by feeding experiments using (13)C-labeled precursors and elucidation of the (13)C-NMR spectrum of (13)C-labeled 2. These studies showed that 2 was derived from five acetates and a methyl group of methionine. In the DPPH-radical scavenging assay, 1 and 2 gave ED(50) values of 149 and 167 muM after standing for 2.0 hr. Compound 2 reacted with the DPPH radical to form reaction product 3 which was determined to be 1-[4-(3,4-dihydro-3-methyl-6-{1,3-pentadienyl}-2,4-dioxo-2H-pyran-3-yl)-phenyl]-1-phenyl-2-picrylhydrazine from spectroscopic evidence.     

Rapid, automated analysis of 13C and 18O of CO2 in gas samples by continuous-flow, isotope ratio mass spectrometry.

A rapid automated method for isotopic analysis of 13C and 18O in CO2 has been developed. A variety of gas samples containing CO2 can be swept from serological tubes into a helium carrier flow; impurities are separated on a GC column so that a pure pulse of CO2 in He flows into the mass spectrometer. Isotopic ratio determinations are carried out as the pulse passes through the mass spectrometer, allowing a sample to be measured approximately every 4 min. A double, concentric needle-probe is used to flush the sample from the tube so that 100% sample recovery is achieved, maximizing sensitivity and preventing the possibility of fractionation. The precision of the technique, sigma(m-1), is better than 0.2% (0.0002 atom per cent excess) for 13C and 0.4% (0.83 p.p.m.) for 18O for 10 micromol of CO2 at natural abundance. Samples containing only atmospheric concentrations of CO2 can also be analyzed.     

Volatile compounds of Asphodelus microcarpus Salzm. et Viv. Honey obtained by HS-SPME and USE analyzed by GC/MS

Chemical analysis of Asphodelus microcarpus Salzm. et Viv. honey is of great importance, since melissopalynology does not allow the unambiguous determination of its botanical origin. Therefore, the volatile compounds of eight unifloral asphodel honeys have been investigated for the first time. The honey extracts were obtained by headspace solid-phase microextraction (HS-SPME) and ultrasonicsolvent extraction (USE) and analyzed by GC and GC/MS. In the honey headspace, 31 volatile compounds were identified with high percentages of 2-phenylacetaldehyde (2; 14.8–34.7%), followed by somewhat lower percentages of methyl syringate (1; 10.5–11.5%). Compound 2 is not a specific marker of the botanical origin of the honey, but its high percentage can be emphasized as headspace characteristic of asphodel honey. The extraction solvent for all the samples was selected after extracting a representative sample with pentane, Et(2)O, pentane/Et(2)O 1:2 (v/v), and CH(2)Cl(2) . Compound 1 was the major constituent of all the USE extracts (46.8–87.0%). According to these preliminary results, all the honey samples were extracted by USE with the solvent pentane/Et(2)O 1:2. A total of 60 volatile compounds were identified with 1 as predominant compound (69.4–87.0%), pointing out 1 as Asphodelus honey volatile marker.     

13C-NMR studies of membrane lipid-protein interactions upon protein heat denaturation.

Spinach chloroplast membranes were studied by natural abundance carbon-13 nuclear magnetic resonance (13C-NMR) spectroscopy in their normal state and after heat denaturation of membrane proteins. The membrane proteins were denatured by raising the temperature of the sample to 67 degrees C for 5 minutes [YashRoy, R.C. (1991) J. Biochem. Biophys. Methods 22, 55-59]. Line-broadening of 13C-NMR resonances arising from the 1st (carbonyl), 7th, 9th and 12th carbon atom of fatty-acyl chains with reference to the carbonyl (C-1) group shows increased immobilization of lipid fatty-acyl chains at these locations, obviously caused by changes in interactions between membrane lipids and proteins upon heat denaturation of membrane proteins.     

Chemoenzymatic synthesis of sucrose-containing aromatic polymers.

A chemoenzymatic approach was developed to prepare sucrose-containing aromatic polymers. The protease from Bacillus licheniformis catalyzed the transesterification of sucrose with a diester of terephthalic acid in pyridine to give the mono- and diester products. At 45 degrees C, >70% of sucrose was consumed after 1 day and sucrose diester began to form after 6 days when >95% of sucrose had been converted to sucrose monoester. The final yield of sucrose diester after 20 days was 13.8%. The sucrose monoester was identified as sucrose 1'-terephthalate and the diester products consisted of sucrose 6,1'-diterephthalate and sucrose 6',1'-diterephthalate in a ratio of 2:1. The sucrose diester products were polymerized with ethylene-glycol and ethylene-diamine to give poly(ethylene-terephthalate) and poly(ethylene-terephthalamide), with sucrose contained in the polymer backbone. The polycondensation reactions were carried out in dimethylsulfoxide (DMSO) at 70 degrees C using zinc acetate as a catalyst. The sucrose-containing polyester and polyamide were obtained at 65% yield for 24 h and at 73% yield for 12 h, respectively. End-group analysis of the polymers by (13)C-NMR or (1)H-NMR in DMSO provided a number average molecular weight of 3200 and 4300 Da, respectively. Structural analyses of the polymers were performed with (1)H-NMR, (13)C-NMR, and FTIR. On the basis of (13)C-NMR, acylation of the C1', C6, and C6' hydroxyls were maintained in the polymer backbones.