Tyrolobibenzyls E and F from Scorzonera humilis and distribution of caffeic acid derivatives,lignans and tyrolobibenzyls in European taxa of the subtribe Scorzonerinae (Lactuceae,Asteraceae)

A chemosystematic study of the subtribe Scorzonerinae, a subtribe of the Lactuceae tribe of the Asteraceae family was performed, using the recently discovered tyrolobibenzyls as well as lignans and caffeic acid derivatives as diagnostic characters. In addition to the known compounds two new tyrolobibenzyls (E and F) were isolated and their structures were established by mass spectrometry and 1D and 2D NMR spectroscopy. Twenty four samples from rootstocks of seventeen different Scorzonerinae taxa, comprising members of three genera (Podospermum, Scorzonera, and Tragopogon), were analyzed. Tyrolobibenzyls A (1), B (2), C (5), D (3), E (6), and F (4) were identified in crude extracts by means of HPLC retention times, on-line UV spectra and on-line MS/MS spectra. Quantification of these compounds was performed by HPLC, using 2,2-bis-(4-hydroxyphenyl)-propane as an internal standard. Tyrolobibenzyls A-F were only detected in samples from Scorzonera humilis, while chlorogenic acid and 3,5-dicaffeoylquinic acid were detected in all samples investigated. In contrast, caffeoyl tartaric acid and cichoric acid were not detectable in any member of the subtribe Scorzonerinae.     

Metabolomic analysis of Strychnos nux-vomica, Strychnos icaja and Strychnos ignatii extracts by 1H nuclear magnetic resonance spectrometry and multivariate analysis techniques

1H Nuclear magnetic resonance spectrometry and multivariate analysis techniques were applied for the metabolic profiling of three Strychnos species: Strychnos nux-vomica (seeds, stem bark, root bark), Strychnos ignatii (seeds), and Strychnos icaja (leaves, stem bark, root bark, collar bark). The principal component analysis (PCA) of the 1H NMR spectra showed a clear discrimination between all samples, using the three first components. The key compounds responsible for the discrimination were brucine, loganin, fatty acids, and Strychnos icaja alkaloids such as icajine and sungucine. The method was then applied to the classification of several "false angostura" samples. These samples were, as expected, identified as S. nux-vomica by PCA, but could not be clearly discriminated as root bark or stem bark samples after further statistical analysis.     

Chemosystematic investigations of irregular diterpenes in Anisotome and related New Zealand Apiaceae

A chemosystematic HPLC-UV and HPLC-MS investigation of New Zealand members of the Apiaceae was performed. Diterpenes were identified and quantified in methanolic extracts from subaerial parts of 28 taxa and 54 samples of Aciphylla, Anisotome, Apium, Gingidia, Lignocarpa, Oreomyrrhis, and Scandia. Six diterpenes (1-2, 4-7) and four polyacetylenes (8-11) were identified. The known compounds were the diterpenes anisotomenoic acid 1, anisotomene-1-ol 2, 16-acetoxyanisotomenoic acid 4 and anisotomene-1,12-diol 5; and the polyacetylenes falcarinol 8, falcarindiol 9, (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 10, and (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 1-acetate 11. New irregular diterpenes 13,14-dihydroanisotom-12E-ene-1,14-diol 6 and 14-methoxy-13,14-dihydroanisotom-12E-ene-1-ol 7 were isolated from A. haastii. Isomers of the new semi-synthetic diterpene 16-hydroxyanisotomenoic acid 3 were detected in extracts of Anisitone flexuosa. Structure elucidation was performed by HR mass spectrometry and 1D and 2D NMR spectroscopy. In crude extracts, compounds were identified by their HPLC retention times and their on-line HPLC-UV and MS spectra. Anisotomene diterpenes occurred in eight out of 16 species of the genus Anisotome, but were not detected in any of the other genera. In contrast, polyacetylenes were present in all the genera investigated.     

Metabolic profiling of Dactylopius (Hemiptera: Dactylopiidae) species pigments by geographical origin and hosts using multivariate data analysis

The genus Dactylopius includes the group of insects historically used in Mexico as a source of natural red colorant, the cochineal color. Five species of Dactylopius collected in thirteen states of Mexico and two provinces of Argentina were analyzed using high-performance liquid chromatography with a photodiode array detector. This analysis allowed each species to be identified on the basis of differences in their metabolic profiles. Cluster analysis and principal component analysis differentiated the species by localities and host plant. These two multivariate data analysis techniques were complementary and confirmed the grouping of all analyzed Dactylopius samples. For all species, carminic acid, identified by reference to a commercial sample, was the major compound present in significant amounts, making all five species potential sources of colorant. In addition, each species could be differentiated by the presence of other minor compounds.     

A high-throughput method for microbial metabolome analysis using gas chromatography/mass spectrometry

An analytical high-throughput method based on gas chromatography/mass spectrometry (GC/MS) was developed for fast metabolome investigation. By parallelization and partial automation the time needed for the preanalytical steps could be reduced. In addition a strong decrease of the relative standard deviation of metabolite concentrations from independent samples on the same microtiter plate from 25 to 13% was achieved. Between different plates the relative standard deviation is comparable to the one observed in standard experiments with shaking flasks. Using a fast GC the time need for the full GC/MS-based metabolome analysis could be decreased from 60 to 18 min per run, allowing the measurement of 72 single samples per day and GC/MS machine. In samples of the model organism Corynebacterium glutamicum more than 1000 peaks in the total ion current could be detected in a single fast GC/MS run of which 650 were strong enough to be quantified. Approximately 150 compounds of these were identified using our metabolite MS-library. Correlation analysis of the concentration vectors of independent wild-type samples raised under the same conditions show very high correlations of 0.99+/-0.01 (logs). In conclusion this method allows screenings of large mutant libraries for genetically induced metabolic perturbations.     

Assessment of 1H NMR spectroscopy and multivariate analysis as a technique for metabolite fingerprinting of Arabidopsis thaliana

An approach to metabolite fingerprinting of crude plant extracts that utilizes 1H nuclear magnetic resonance (NMR) spectroscopy and multivariate statistics has been tested. Using ecotypes of Arabidopsis thaliana as experimental material, a method has been developed for the rapid analysis of unfractionated polar plant extracts, enabling the creation of reproducible metabolite fingerprints. These fingerprints could be readily stored and compared by a variety of chemometric methods. Comparison by principal component analysis using SIMCA-P allowed the generation of residual NMR spectra of the compounds that contributed significantly to the differences between samples. From these plots, conclusions were drawn with respect to the identity and relative levels of metabolites differing between samples.     

Chemical variability within the marine sponge Aplysina fulva

Dibromotyrosine-derived metabolites are of common occurrence within marine sponges belonging to the order Verongida. However, previous chemical analysis of crude extracts obtained from samples of the verongid sponge Aplysina fulva collected in Brazil did not provide any dibromotyrosine-derived compounds. In this investigation, five samples of A. fulva from five different locations along the Brazilian coastline and one sample from a temperate reef in the South Atlantic Bight (SAB) (Georgia, USA) were investigated for the presence of bromotyrosine-derived compounds. All six samples collected yielded dibromotyrosine-derived compounds, including a new derivative, named aplysinafulvin, which has been identified by analysis of spectroscopic data. These results confirm previous assumptions that dibromotyrosine-derived metabolites can be considered as chemotaxonomic markers of verongid sponges. The isolation of aplysinafulvin provides additional support for a biogenetic pathway involving an arene oxide intermediate in the biosynthesis of Verongida metabolites. It cannot yet be established if the chemical variability observed among the six samples of A. fulva collected in Brazil and the SAB is the result of different environmental factors, distinct chemical extraction and isolation protocols, or a consequence of hidden genetic diversity within the postulated morphological plasticity of this species.     

Malonylated flavonol glycosides from the petals of Clitoria ternatea

Three flavonol glycosides, kaempferol 3-O-(2"-O-alpha-rhamnosyl-6"-O-malonyl)-beta-glucoside, quercetin 3-O-(2"-O-alpha-rhamnosyl-6"-O-malonyl)-beta-glucoside, and myricetin 3-O-(2",6"-di-O-alpha-rhamnosyl)-beta-glucoside were isolated from the petals of Clitoria ternatea cv. Double Blue, together with eleven known flavonol glycosides. Their structures were identified using UV, MS, and NMR spectroscopy. They were characterized as kaempferol and quercetin 3-(2(G)- rhamnosylrutinoside)s, kaempferol, quercetin, and myricetin 3-neohesperidosides, 3-rutinosides, and 3-glucosides in the same tissue. In addition, the presence of myricetin 3-O-(2"-O-alpha-rhamnosyl-6"-O-malonyl)-beta-glucoside was inferred from LC/MS/MS data for crude petal extracts. The flavonol compounds identified in the petals of C. ternatea differed from those reported in previous studies.     

新疆雪莲的高效液相指纹图谱及液-质联用分析

建立新疆雪莲药材的高效液相指纹图谱。以Luna C18为分析柱,用乙腈-0.1%醋酸梯度洗脱,获得分离度较好的新疆雪莲药材HPLC分析条件。通过10批样品的分析和对照,标示出12个共有峰,相似度分析大于0.92。最后通过HPLC-UV-MS/MS联用分析鉴定出其中7个化学成分。该方法可为新疆雪莲药材指纹图谱和质量分析方法的建立提供参考。     

Novel dermaseptins from Phyllomedusa hypochondrialis (Amphibia)

Six new antimicrobial peptides structurally related to the dermaseptin family have been isolated from the skin secretion of the amphibian Phyllomedusa hypochondrialis. The primary structures of these molecules named as DShypo 01, 02, 03, 04, 06, and 07 were determined by de novo MS/MS experiments, Edman degradation, and cDNA sequencing. The fifth peptide was found to be precisely the same DS 01 from Phyllomedusa oreades previously described by our group. The majority of the peptides purified from the crude skin secretion could be directly localized and mapped onto a freshly dissected dorsal skin fragment using mass spectrometry-imaging techniques. Comparisons between peptides and commercial drugs on their antibacterial and anti-Leishmania amazonensis efficiencies, associated with peptide lytic effects on mammalian blood cells and surface plasmon resonance interaction studies on immobilized DMPC vesicles, were also performed.     

Sesquiterpene constituents from the essential oil of the liverwort Plagiochila asplenioides

The essential oil of the liverwort Plagiochila asplenioides from two different locations in Northern Germany were investigated by chromatographic and spectroscopic methods. Seven compounds were isolated by preparative gas chromatography (GC) and their structures investigated by mass spectrometry (MS), NMR techniques and chemical correlations in combination with enantioselective GC. In addition to known constituents, aromadendra-1(10),3-diene, two aromatic sesquiterpene hydrocarbons, bisabola-1,3,5,7(14)-tetraene and bisabola-1,3,5,7-tetraene, three sesquiterpene ethers, muurolan-4,7-peroxide, plagiochilines W and X, in addition to ent-4-epi-maaliol, could be identified as natural compounds for the first time.     

The chemotaxonomic classification of Korean Campanulaceae based on triterpene,sterol, and polyacetylene contents

The taxonomic usefulness of selected marker compounds was investigated by analyzing the chemotaxonomy of 25 taxa in the Korean Campanulaceae. Our data permit discrimination of the source plants of crude drugs listed in the Korean Pharmacopoeia and the Korean Herbal Pharmacopoeia. Chemotaxonomic analysis methods were validated, and quantitative measurements of six marker compounds were made using HPLC. Marker compound similarities among taxa were identified through multivariate statistical analyses (principal component analysis and hierarchical cluster analysis). The chemical analysis method was validated with regard to linearity, lower limit of detection, lower limit of quantitation, precision, accuracy, and recovery. The analysis revealed differences in the marker compound composition of each genus. Eight genera comprising Adenophora, Codonopsis, Asyneuma, Campanula, Hanabusaya, Platycodon, Wahlenbergia, and Peracarpa clustered according to the chemical classification. The results indicated that the six marker compounds used in this analysis were useful in identifying Korean Campanulaceae. These marker compound data were able to successfully discriminate among the three species that are sold as the crude drug Adenophorae Radix.     

Fingerprint profile of Ginkgo biloba nutritional supplements by LC/ESI-MS/MS

Ginkgo biloba is one of the most popular herb nutrition supplements, with terpene lactones and flavonoids being the two major active components. A fingerprint profile method was developed using a capillary HPLC/MS method which can identify more than 70 components from the G. biloba product. The method allows the flavonoids and terpene lactones to be detected simultaneously and information of both the parent ion and its fragmentation can be obtained in just one HPLC/MS run. Targeted post-acquisition analysis allows mass spectrometric information regarding the identification of flavonoid components to be easily distinguished from other data, however the same approach for terpene lactones was less successful due to dimer formation and requires further development. The fingerprint profiles of five commercial G. biloba nutritional supplements were obtained and compared; variation of some components among the samples was observed and fortification could be detected. In the quality control analysis of the G. biloba product this method could be viewed as complementary to specific quantitative analysis of some bioactive components of the herb.     

Chemotaxonomy of the tribe Antidesmeae (Euphorbiaceae): antidesmone and related compounds

Selected species of the tribe Antidesmeae (Euphorbiaceae, subfamily Phyllanthoideae) have been screened for antidesmone occurrence and its content by quantitative HPLC (UV) and qualitative LC-MS/MS analysis. The LC-MS analysis allowing the additional detection of 17,18-bis-nor-antidesmone, 18-nor-antidesmone, 8-dihydroantidesmone and 8-deoxoantidesmone was carried out in the selected reaction monitoring (SRM) mode. Leaf material from herbarium specimens of 13 Antidesma spp., Hyeronima alchorneoides and Thecacoris stenopetala (all subtribe Antidesminae), as well as Maesobotrya barteri, Aporosa octandra (both Scepinae) and Uapaca robynsii (Uapacinae) were analysed. Additionally, freshly collected samples of different plant parts of two Antidesma spp. were investigated to ensure the significance of the results on herbarium specimens and to compare the antidesmone content in bark, root and leaves. Antidesmone could be unambiguously identified in 12 of 13 Antidesma spp., as well as in the two other investigated genera of subtribe Antidesminae, in levels of up to 65 mg/kg plant dry weight. Antidesmone was not found in specimens from other subtribes. Antidesmone-derived compounds occur in much lower concentrations than antidesmone.     

Volatile constituents in the liverwort Tritomaria polita

The essential oil of the liverwort Tritomaria polita, collected in Otztal/Tyrol (Austria), was investigated by chromatographic and spectroscopic methods. Several new compounds were isolated by preparative gas chromatography (GC) and their structures investigated by mass spectrometry (MS) and NMR techniques. In addition to known constituents, the sesquiterpenoids (+)-eudesma-3,11-dien-8-one, (+)-eudesma-3,7(11)-dien-8-one, (+)-6,11-epoxy-eudesmane, (-)-6,7-seco-eudesm-7(11)-en-6-al, (+)-6beta-hydroxy-eudesm-11-ene, (-)-6alpha-hydroxy-eudesm-11-ene, (+)-6,11-epoxy-isodaucane could be identified as natural compounds for the first time.     

Analysis of volatiles from black pine (Pinus nigra): significance of wounding and egg deposition by a herbivorous sawfly

The composition of headspace volatiles of black pine (Pinus nigra) was analysed by coupled gas chromatography-mass spectrometry (GC-MS). It has been shown in a previous study that egg deposition of the sawfly Diprion pini on P. sylvestris induces a quantitative change of the pine volatile blend . Chrysonotomyia ruforum, an egg parasitoid of D. pini, is known to be attracted by volatiles from egg-carrying P. sylvestris, but not by odour from egg-laden P. nigra. Therefore, the present study focused on the question whether also P. nigra as another host plant of this sawfly responds to egg deposition by change of its volatile blend. The headspace of untreated, egg-carrying, and artificially wounded P. nigra twigs were compared. The artificial damage inflicted to the twigs mimicked the damage by the sawfly female prior to egg deposition. Thirty five mainly terpenoid compounds that were identified in more than 50% of the egg-carrying P. nigra twigs could also be detected in the headspace of untreated and artificially wounded twigs. Quantitative differences of the blends of volatiles from differently treated P. nigra twigs were compared by multivariate data analyses. PLS-DA (projection to latent structures-discriminant analysis) revealed that blends of volatiles from differently treated P. nigra differed significantly. When comparing volatiles from egg-carrying and artificially wounded P. nigra with respective P. sylvestris samples, qualitative and quantitative differences were detected. The differences in volatile composition of P. nigra and P. sylvestris are discussed with special respect to the egg parasitoid's response to odours of egg-carrying pine twigs.     

Bafoudiosbulbins A, and B, two anti-salmonellal clerodane diterpenoids from Dioscorea bulbifera L. var sativa

Two clerodane diterpenoids, Bafoudiosbulbins A 1, and B 2, together with five known compounds: tetracosanoic acid, 1-(tetracosanoyl)-glycerol, trans-tetracosanylferulate, beta-sitosterol and 3-O-beta-D-glucopyranosyl-beta-sitosterol were isolated from the tubers of Dioscorea bulbifera L. var sativa. Their structures were established by spectroscopic methods (1D and 2D-NMR, MS) and X-ray crystallographic diffraction analysis of compound 1. The CH2Cl2-soluble portion of the crude extract and the two clerodanes were screened for anti-bacterial activity using both agar diffusion and broth dilution techniques against Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Salmonella typhi, Salmonella paratyphi A and Salmonella paratyphi B. They both showed significant activities against P. aeruginosa, S. typhi, S. paratyphi A and S. paratyphi B.     

毛冬瓜根挥发油化学成分分析

采用水蒸气蒸馏法提取,运用毛细管气相色谱/质谱联用法首次对毛冬瓜根挥发油化学成分进行了分析研究,确认了其中的49种化合物,用气相色谱面积归一法测定各组分的相对百分含量.在分离出的主要挥发性成分中含有烃类(22.26%)、醛酮类(8.52%)、醇类(45.38%)、酚类(1.08%)、羧酸类(7.69%)、酯类(7.18%)、杂环类(2.72%)和环氧类(3.39%)等8大类.在主要成分中以萜类化合物为主(62.20%),包括5种单萜(5.29%)和17种倍半萜(56.91%).其中多种成分为已知药用成分,为进一步评价其质量和开发新药提供了基础数据.