RESUMO
Commercially available linear alkylbenzenesulfonates (LASs) are a mixture of various homologues and isomers, leading to 20 major species. In this work we investigated the commercial product by liquid chromatography-solid phase extraction-nuclear magnetic resonance spectroscopy-mass spectrometry (LC-SPE-NMR/MS). The commercial product was separated into 17 fractions by liquid chromatography (LC). After chromatographic separation, 5% of the flow was split to a mass spectrometer (MS) while 95% was send to post-column solid phase extraction cartridges for enrichment of the analytes (LC-SPE). After elution from the SPE-cartridges a NMR-spectrometer equipped with a cryo-probe was used for the characterisation of the different LASs species. For the first time (1)H-1D and H-H-COSY spectra for 14 LASs species out of 20 major isomers are presented, whereas the 6 remaining species are detected as mixtures in 3 (1)H-1D and H-H-COSY spectra. These data were used to correlate the chromatographic retention of the LASs isomers to the substitution pattern of the alkyl chain.
Assuntos
Envelhecimento da Pele/fisiologia , Dermatopatias/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Humanos , Lipídeos de Membrana/metabolismo , Pessoa de Meia-Idade , Pele/fisiopatologia , Dermatopatias/fisiopatologia , Dermatopatias/prevenção & controle , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
For rapid screening of natural products from Actinomycetes, a combination of on-line couplings LC-NMR, LC-DAD-MS and HPLC-PDA, as well as MALDI-TOF-MS is particularly suitable. Simultaneous use of these coupling techniques provides considerable advantages for the rapid identification of natural compounds in mixtures. The results of our present investigation on secondary metabolite products of Streptomyces violaceoruber TU 22 showed that more than 50% of the identified metabolites are new compounds. The structures of four new polyketides (granaticin C, metenaticin A, B and C) as well as four known ones (granaticin A, granatomycin E, daidzein and genistein) have been elucidated using LC-NMR, LC-MS/MS and -MS(n) techniques in combination with two-dimensional NMR spectroscopy.
Assuntos
Fatores Biológicos/análise , Cromatografia Líquida/métodos , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Streptomyces/química , Fatores Biológicos/química , Genisteína/análise , Isoflavonas/análise , Macrolídeos/análise , Naftoquinonas/análise , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Streptomyces/metabolismoRESUMO
Nodular thyroid disease is highly prevalent in iodine deficient areas. In Germany it affects approximately 30 % of the adult population. Differential diagnosis of thyroid nodules is directed at exclusion of rare thyroid malignancy and assessment of the nodules' functional characteristics in order to determine the best treatment approach. In 2003 the annual meeting of the Sektion Schilddrüse, a thyroid specialist group of the German Society of Endocrinology addressed the topic of "management of benign nodular thyroid disease". To assess the current diagnostic and therapeutic approach to nodular thyroid disease by German thyroid experts we designed a questionnaire, which was sent to all members of the Thyroid Section before the meeting and was answered by 70 % of the participants. We here report the results of the questionnaire, which show a considerable national heterogeneity in the management of thyroid nodules as well as diagnostic and treatment preferences for different thyroid specialist subgroups, e.g. endocrinologists and nuclear medicine specialist. This is in agreement with results of previous questionnaires assessing state-of-the-art practise among members of the European and American Thyroid Associations and underlines the definite need for a consensus process and for carefully planned randomized trials to answer the many unresolved questions in diagnosis and therapy of nodular thyroid disease.
Assuntos
Inquéritos e Questionários , Nódulo da Glândula Tireoide/diagnóstico , Nódulo da Glândula Tireoide/terapia , Biópsia por Agulha , Endocrinologia , Seguimentos , Alemanha/epidemiologia , Bócio Nodular/diagnóstico , Humanos , Incidência , Iodo/uso terapêutico , Cintilografia , Sociedades Médicas , Nódulo da Glândula Tireoide/epidemiologia , Tiroxina/uso terapêuticoRESUMO
A new cycloartane glycoside (1) was obtained from a minor triterpene fraction of the rhizome extract of Actaea racemosa (synonym: Cimicifuga racemosa) along with a known compound, cimigenol 3-O-beta-D-xylopyranoside. The structure of 1 was elucidated as 20(S),22(R),23(R),24(S)-12beta-acetoxy-16beta:23,23alpha:24-diepoxy-3beta,22beta,25-trihydroxy-9,19-cyclolanost-7-ene 3-O-beta-D-xylopyranoside (actaeaepoxide 3-O-beta-D-xylopyranoside) on the basis of spectral and chemical evidence.
Assuntos
Glicosídeos/isolamento & purificação , Plantas Medicinais/química , Triterpenos/isolamento & purificação , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Compostos de Epóxi/química , Glicosídeos/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Raízes de Plantas/química , Espectrometria de Massas por Ionização por Electrospray , Triterpenos/químicaRESUMO
We have synthesized and fully characterized the NK(2) receptor antagonist nepadutant and its by-products using nuclear magnetic resonance (NMR) and restrained molecular dynamics. The agent consists of an active bicyclic hexapeptide combined with a sugar residue. Analysis of the high-performance liquid chromatogram and the mass spectroscopy spectra yields traces of three by-products with the same molecular weight as the main product. The conformation of the molecules in the bicyclic hexapeptide segment, the active region, is well defined, whereas the sugar moiety is disordered. For the peptide region of nepadutant and all of its by-products, the NMR observables can be described by a single backbone conformation, more specifically a betaI, betaII-turn arrangement. The active dipeptide unit Trp-Phe occupies the i+1 and i+2 position of a betaI-turn. The by-product profile is characterized by different forms of sugars which are caused mainly by isomerization in the process of ring opening.
Assuntos
Glicopeptídeos/química , Peptídeos Cíclicos/química , Receptores da Neurocinina-2/antagonistas & inibidores , Cromatografia Líquida de Alta Pressão , Glucose/química , Glicopeptídeos/síntese química , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Conformação Molecular , Peso Molecular , Peptídeos Cíclicos/síntese química , Conformação Proteica , Estrutura Secundária de ProteínaRESUMO
The twigs of Rhus pyroides yielded a novel bichalcone 2',4",2"'-trihydroxy-4',4"'-dimethoxy-4-O-5"'-bichalcone. It was identified on the basis of spectroscopic data including 1D and 2D NMR spectroscopy. The name rhuschalcone-1 is proposed.
Assuntos
Chalcona/análogos & derivados , Plantas Tóxicas , Toxicodendron/química , Chalcona/química , Chalcona/isolamento & purificação , Chalconas , Espectroscopia de Ressonância Magnética , Caules de Planta/químicaRESUMO
LC/NMR and LC/MS (the latter technique in the MSn mode) were used to characterize the organic constituents of industrial wastewater with emphasis on polar, nonvolatile compounds. In the effluent of a textile company, various compounds such as anthraquinone-type dyes and their byproducts, a fluorescent brightener, a byproduct from polyester production, and auxiliaries such as anionic and nonionic surfactants and their degradation products were identified. It is shown that the combined use of both hyphenated techniques provides complementary structural information. If applied under comparable chromatographic conditions, they are well-suited for the nontarget analysis.
RESUMO
The solution structure of eight cyclic pentapeptides has been determined by two-dimensional 1H-NMR spectroscopy combined with spectra simulations and restrained molecular dynamic simulations. Six of the cyclic pentapeptides were derived from the C-terminal cholecystokinin fragment CCK-4 enlarged with Asp1 resulting in the sequence (Asp-Trp-Met-Asp-Phe), one L-amino acid after the other was substituted by its D-analog. In addition, two peptides, including an all-L-amino-acid-containing cyclic pentapeptide, cyclo(Asp-Phe-Lys-Ala-Thr) and cyclo(Asp-Phe-Lys-Ala-D-Thr) were investigated. All D-amino-acid-containing peptides show beta II'-turn conformations with the D-amino acid in the i + 1 position, excepting the D-aspartic-acid-containing peptides. These two peptides are characterized by the lack of beta-turns at pH values less than 4, suggesting that D-aspartic acid in the full-protonized state avoids the formation of beta-turns in these compounds. At pH values greater than 5, a conformational change into the beta II'-turn conformation was also observed for these peptides. Conformations without beta-turns are expected for cyclic all-L pentapeptides, but both cyclo(Asp-Phe-Lys-Ala-Thr) and the D-Thr analog cyclo(Asp-Phe-Lys-Ala-D-Thr) exhibit beta II'-turn conformations around Thr-Asp and D-Thr-Asp. Thus cyclic all-L pentapeptides and those with one D-amino acid are able to form similar structures preferably with a beta II'-turn. The beta-turn formation in cyclic pentapeptides containing a D-aspartic acid is dependent on the ionization state. The relevance of the work to the design of beta'-turn mimetics is discussed.
Assuntos
Peptídeos Cíclicos/química , Estrutura Secundária de Proteína , Ligação de Hidrogênio , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Molecular , Fragmentos de Peptídeos/química , Tetragastrina/químicaRESUMO
Two methods for the quantitative determination of compounds in continuous-flow HPLC/NMR are described. The first method uses an internal standard (caffeine) of known concentration directly mixed into the mobile phase, while with the second method, a known amount of internal standard is injected onto the column during the chromatographic run. The latter method was validated using several nitroaromatic compounds and explosives. Deviations between the injected and calculated amounts of analytes are usually below 10% while the relative standard deviation ranges from 2% in the upper microgram range to 40% at the limit of detection.
RESUMO
An aqueous solution of 2,4,6-trinitrotoluene (TNT) was irradiated by natural sunlight for a period of 1 month to generate phototransformation products of this compound. After solid-phase extraction on a poly(styrene-divinylbenzene) copolymer at pH 1, the structures of several acidic nitroaromatic compounds were identified by means of continuous-flow HPLC/(1)H NMR and HPLC/TSP-MS measurements of this extract. By interpretation of both NMR and MS spectra, it was even possible to characterize noncommercially available phototransformation products of TNT. The results obtained by continuous-flow HPLC/(1)H NMR were compared with those obtained by the investigation of a groundwater sample of a former ammunition site near Elsnig, Germany. The results show that several identified phototransformation products of TNT are also present in this groundwater sample.
RESUMO
Coupling of HPLC to NMR was applied for the first time to the analysis of environmental samples, i.e., water samples from an ammunition hazardous waste site. Using the continuous flow mode at very low flow rates (< or = 0.017 mL/min) and large volume injection (400 microL), the confirmation of many nitroaromatic compounds could be achieved down to the microgram-per-liter level after solid phase extraction of a groundwater sample from a former ammunition production site. At a flow rate of 0.006 mL/min, it is possible to detect less than 29 nmol (5 micrograms) of 1,3-dinitrobenzene injected on a 75 mm x 4 mm reversed phase C-18 column (particle size, 5 microns). The results obtained by HPLC-NMR are compared to those obtained by HPLC-PDA (photodiode array) of the same sample, demonstrating that many more compounds can be identified by the former compared to the latter method as a result of coelution of major and minor components in the HPLC chromatogram.
Assuntos
Água Doce/química , Resíduos Perigosos , Poluentes Químicos da Água/análise , Cromatografia Líquida de Alta Pressão , Indústrias , Espectroscopia de Ressonância MagnéticaRESUMO
2D 1H NMR spectroscopy of two alpha-helical peptides which differ in their amphipathicity has been used to investigate the relationships between amide-proton chemical shifts, amide-proton exchange rates, temperature, and trifluoroethanol (TFE) concentration. In 50% TFE, in which the peptides are maximally helical, the amide-proton chemical shift and temperature coefficient patterns are very similar to each other in each peptide. Temperature coefficients from -10 to -6 ppb/K, usually indicative of the lack of intramolecular hydrogen bonds, were observed even for hydrophobic amino acids in the center of the alpha-helices. However, slow hydrogen isotope exchange for residues from 4 to 16 in both 18-mer helices indicates intact intramolecular hydrogen bonds over most of the length of these peptides. Based on these anomalous observations, we suggest that the pattern of amide-proton shifts in alpha-helices in H20/TFE solvents is dominated by bifurcated intermolecular hydrogen-bond formation between the backbone carbonyl groups and TFE. The amide-proton chemical shift changes with increasing temperature may be interpreted by a disruption of intermolecular hydrogen bonds between carbonyl groups and the TFE in TFE/water rather than by the length of intramolecular hydrogen bonds in alpha-helices.
Assuntos
Espectroscopia de Ressonância Magnética , Peptídeos/química , Trifluoretanol/química , Hidrogênio , Dobramento de Proteína , TemperaturaRESUMO
The cyclic, imido acid containing pentapeptides cyclo(Asp-Trp-(NMe)Nle-Asp-Phe) (cpp[NMeNle(3)]) and cyclo(Asp-Trp-Pro-Asp-Phe) (cpp[Pro(3)]) have been investigated by 1H-NMR spectroscopy in DMSO and by restrained molecular dynamics methods. The spectra indicate the existence of at least four cis/trans isomers for cpp[NMeNle(3)] and two cis/trans isomers for cpp[Pro(3)]. In addition to the imido peptide bonds, cpp[NMeNle(3)] shows cis/trans isomerization of the Asp4-Phe5 and Phe5-Asp1 peptide bonds whereas only the Phe5-Asp1 peptide bond isomerizes in the Pro-containing peptide. In cpp[Pro(3)] all cis bonds are centred in betaVIb turns. Also, cpp[NMeNle(3)] prefers backbone angles around the cis bonds which are rather similar to the angles of a betaVIb turn. The higher number of cis/trans isomers and slight deviations in the backbone angles of comparable isomers of both peptides are caused by an enhanced flexibility of cpp[NMeNle(3)] due to the possibility of the phi-(NMe)Nle rotation.
Assuntos
Norleucina/química , Peptídeos Cíclicos/química , Prolina/química , Sequência de Aminoácidos , Espectroscopia de Ressonância Magnética , Dados de Sequência Molecular , Oligopeptídeos/química , Conformação ProteicaRESUMO
The conformational behavior of cyclic peptides of the amino acid sequence Cys-Phe/Ala-Pro-Ala-Cys has been investigated through the combined use of molecular simulation methods and NMR experiments to find models for beta-VIa turns of proteins. Both oxidized (cyclic) peptides and reduced (linear) forms were investigated. At least 95% of the cyclic peptides show a cis conformation of the Xaa-Pro bond in solution in DMSO or water, whereas all other peptide bonds are trans. Furthermore, we observed a hydrogen bond between the NH group of residue Ala4 and the C = O group of residue Cys1. Both properties are indicative of beta-VIa turns. After reduction of the disulfide bridge, the all-trans form of the peptide bonds predominates.
Assuntos
Peptídeos/química , Conformação Proteica , Sequência de Bases , Dissulfetos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Dados de Sequência MolecularRESUMO
The conformational analysis of the CCK-B binding peptide cyclo (Asp-Trp-Met-Asp-Phe) has been carried out in DMSO-d6 and in a mixture of H2O/DMSO-d6 by NMR spectroscopy and by restrained molecular dynamics methods. In the NMR spectra, only one set of resonance signals was found. The NOE analyses proved the existence of an all-trans conformation for this peptide. Distance constraints of 1H pairs derived from NOE data were used for restrained molecular dynamics simulations, resulting in one conformational family with a very regular orientation of the amino acids and similar dihedral angles for each residue. The dihedrals and the absence of an intramolecular hydrogen bond indicate that there is no common turn formation in the peptide backbone. A submicromolar binding constant for CCK-B receptors point to a similarity with the bioactive conformation.
Assuntos
Colecistocinina/análogos & derivados , Peptídeos Cíclicos/química , Sequência de Aminoácidos , Fenômenos Químicos , Físico-Química , Dimetil Sulfóxido , Espectroscopia de Ressonância Magnética , Dados de Sequência Molecular , Conformação Proteica , Prótons , SoluçõesRESUMO
A new procedure has been developed for investigating the ability of paramagnetic metal complexes to penetrate the plasma membrane of eukaryotic cells without decomposition. Defolliculated Xenopus laevis oocytes formed the biological system to test N,N-ethylenebis-(1,5,5-trimethyltetramic-acid-3-acetiminato) copper (II). An increase of the signal intensities in spin-echo (SE) images of oocytes treated with the tested substance indicated that the complex was able to penetrate biological membranes due to the arrangement of hydrophobic and hydrophilic groups within the ligand. In contrast, the treatment with the commonly used contrast agent gadolinium-DTPA/dimeglumine did not enhance the signal intensity in NMR images of oocytes after time periods of exposure comparable to those used for the copper complex. After microinjection into Xenopus oocytes the copper complex was released into the extracellular medium without degradation, as shown by HPLC measurements.
Assuntos
Permeabilidade da Membrana Celular , Espectroscopia de Ressonância Magnética , Compostos Organometálicos/farmacocinética , Animais , Cromatografia Líquida de Alta Pressão , Meios de Contraste/farmacocinética , Microscopia , Oócitos/citologia , Oócitos/metabolismo , XenopusRESUMO
The thermally induced decomposition of disaccharide Amadori compounds has been compared to those of monosaccharide ones under almost water-free conditions. The structure of the synthesized maltulosyl compound has been proved to be 4C1-alpha-D-glucopyranosyl- (1----4)-2C5-beta-D-fructopyranosylglycine by 1H- and 13C-NMR spectroscopy. The decomposition of Amadori compounds has been used to study the kinetics of the browning reaction. Compared to fructosylglycine and maltotriulosylglycine, the browning of the disaccharide is faster. Curie point pyrolysis at 300 degrees C and investigation of the pyrolysate by gas chromatography/mass spectrometry have shown that the disaccharide component influences the thermal process. Furanes and furanones have been detected as predominant degradation products, the main one being 2(5H)-furanone. For the first time, we suggest a reaction pathway for the formation of these products via the Maillard reaction which includes 1,6-anhydroglucose.