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Good Resolution (good + resolution)
Selected AbstractsField-amplified sample injection-micellar electrokinetic capillary chromatography for the analysis of bisphenol A, bisphenol F, and their diglycidyl ethers and derivatives in canned soft drinksELECTROPHORESIS, Issue 9 2010Héctor Gallart-Ayala Abstract Conditions were established for the separation and analysis of bisphenol A, bisphenol F, and their diglycidyl ethers by micellar electrokinetic capillary chromatography (MECC). Good resolution was obtained for all compounds, although in order to achieve the separation of ortho,ortho, ortho,para, and para,para isomers of bisphenol F diglycidyl ether (BFDGE), BFDGE·2H2O and BFDGE·2HCl, it was necessary to use a 25,,m id fused silica capillary. To increase sensitivity, a field-amplified sample injection (FASI)-MECC method was developed using 10,mM SDS solution as injection matrix and a 75,,m id fused silica capillary. Instrumental quality parameters such as LODs (<55,,g/L with standards), linearity (r2>0.999), and run-to-run and day-to-day precisions (RSD values lower than 12.5%) were determined. Finally, the suitability of the FASI-MECC method for the analysis of bisphenol A, bisphenol F, and their diglycidyl ethers in canned soft drinks was evaluated. Quantitation was performed by matrix-matched calibration using a plastic-bottled isotonic drink as matrix. The results showed that FASI-MECC is an economic method for the screening and quantitation of these kinds of compounds in soft drink beverages, with no loss of reproducibility, and effective at concentrations lower than the specific migration level values established by the European Union. [source] Influence of microemulsion chirality on chromatographic figures of merit in EKC: Results with novel three-chiral-component microemulsions and comparison with one- and two-chiral-component microemulsionsELECTROPHORESIS, Issue 17 2007Kimberly A. Kahle Abstract Novel microemulsion formulations containing all chiral components are described for the enantioseparation of six pairs of pharmaceutical enantiomers (atenolol, ephedrine, metoprolol, N -methyl ephedrine, pseudoephedrine, and synephrine). The chiral surfactant dodecoxycarbonylvaline (DDCV, R - and S -), the chiral cosurfactant S -2-hexanol, and the chiral oil diethyl tartrate (R - and S -) were combined to create four different chiral microemulsions, three of which were stable. Results obtained for enantioselectivity, efficiency, and resolution were compared for the triple-chirality systems and the single-chirality system that contained chiral surfactant only. Improvements in enantioselectivity and resolution were achieved by simultaneously incorporating three chiral components into the aggregate. The one-chiral-component microemulsion provided better efficiencies. Enantioselective synergies were identified for the three-chiral-component nanodroplets using a thermodynamic model. Additionally, two types of dual-chirality systems, chiral surfactant/chiral cosurfactant and chiral surfactant/chiral oil, were examined in terms of chromatographic figures of merit, with the former providing much better resolution. The two varieties of two-chiral-component microemulsions gave similar values for enantioselectivity and efficiency. Lastly, the microemulsion formulations were divided into categories based on the number of chiral microemulsion reagents and the average results for each pair of enantiomers were analyzed for trends. In general, enantioselectivity and resolution were enhanced while efficiency was decreased as more chiral components were used to create the pseudostationary phase (PSP). [source] Sensitive and simultaneous analysis of five transgenic maizes using multiplex polymerase chain reaction, capillary gel electrophoresis, and laser-induced fluorescenceELECTROPHORESIS, Issue 14 2004Virginia García-Cañas Abstract The benefits of using multiplex polymerase chain reaction (PCR) followed by capillary gel electrophoresis with laser-induced fluorescence (CGE-LIF) for the simultaneous detection of five transgenic maizes (Bt11, T25, MON810, GA21, and Bt176) are demonstrated. The method uses a hexaplex PCR protocol to amplify the five mentioned transgenic amplicons plus the zein gene used as reference, followed by a CGE-LIF method to analyze the six DNA fragments. CGE-LIF was demonstrated very useful and informative for optimizing multiplex PCR parameters such as time extension, PCR buffer concentration and primers concentration. The method developed is highly sensitive and allows the simultaneous detection in a single run of percentages of transgenic maize as low as 0.054% of Bt11, 0.057% of T25, 0.036% of MON810, 0.064% of GA21, and 0.018% of Bt176 in flour obtaining signals still far from the detection limit (namely, the signal/noise ratios for the corresponding DNA peaks were 41, 124, 98, 250, 252, and 473, respectively). These percentages are well below the minimum threshold marked by the European Regulation for transgenic food labeling (i.e., 0.5,0.9%). A study on the reproducibility of the multiplex PCR-CGE-LIF procedure was also performed. Thus, values of RSD lower than 0.67 and 6.80% were obtained for migration times and corrected peak areas, respectively, for the same sample and three different days (n = 12). On the other hand, the reproducibility of the whole procedure, including four different multiplex PCR amplifications, was determined to be better than 0.66 and 23.3% for migration times and corrected peak areas, respectively. Agarose gel electrophoresis (AGE) and CGE-LIF were compared in terms of resolution and sensitivity for detecting PCR products, demonstrating that CGE-LIF can solve false positives induced by artifacts from the multiplex PCR reaction that could not be addressed by AGE. Moreover, CGE-LIF provides better resolution and sensitivity. To our knowledge, these results demonstrate for the first time that multiplex PCR-CGE-LIF is a solid alternative to determine multiple genetically modified organisms in maize flours in a single run. [source] Eukaryotic diversity and phylogeny using small- and large-subunit ribosomal RNA genes from environmental samplesENVIRONMENTAL MICROBIOLOGY, Issue 12 2009William Marande Summary The recent introduction of molecular techniques in eukaryotic microbial diversity studies, in particular those based in the amplification and sequencing of small-subunit ribosomal DNA (SSU rDNA), has revealed the existence of an unexpected variety of new phylotypes. The taxonomic ascription of the organisms bearing those sequences is generally deduced from phylogenetic analysis. Unfortunately, the SSU rDNA sequence alone has often not enough phylogenetic information to resolve the phylogeny of fast-evolving or very divergent sequences, leading to their misclassification. To address this problem, we tried to increase the phylogenetic signal by amplifying the complete eukaryotic rDNA cluster [i.e. the SSU rDNA, the internal transcribed spacers, the 5.8S rDNA and the large-subunit (LSU) rDNA] from environmental samples, and sequencing the SSU and LSU rDNA part of the clones. Using marine planktonic samples, we showed that surveys based on either SSU or SSU + LSU rDNA retrieved comparable diversity patterns. In addition, phylogenetic trees based on the concatenated SSU + LSU rDNA sequences showed better resolution, yielding good support for major eukaryotic groups such as the Opisthokonta, Rhizaria and Excavata. Finally, highly divergent SSU rDNA sequences, whose phylogenetic position was impossible to determine with the SSU rDNA data alone, could be placed correctly with the SSU + LSU rDNA approach. These results suggest that this method can be useful, in particular for the analysis of eukaryotic microbial communities rich in phylotypes of difficult phylogenetic ascription. [source] High-order ENO and WENO schemes for unstructured gridsINTERNATIONAL JOURNAL FOR NUMERICAL METHODS IN FLUIDS, Issue 10 2007W. R. Wolf Abstract This work describes the implementation and analysis of high-order accurate schemes applied to high-speed flows on unstructured grids. The class of essentially non-oscillatory schemes (ENO), that includes weighted ENO schemes (WENO), is discussed in the paper with regard to the implementation of third- and fourth-order accurate methods. The entire reconstruction process of ENO and WENO schemes is described with emphasis on the stencil selection algorithms. The stencils can be composed by control volumes with any number of edges, e.g. triangles, quadrilaterals and hybrid meshes. In the paper, ENO and WENO schemes are implemented for the solution of the dimensionless, 2-D Euler equations in a cell centred finite volume context. High-order flux integration is achieved using Gaussian quadratures. An approximate Riemann solver is used to evaluate the fluxes on the interfaces of the control volumes and a TVD Runge,Kutta scheme provides the time integration of the equations. Such a coupling of all these numerical tools, together with the high-order interpolation of primitive variables provided by ENO and WENO schemes, leads to the desired order of accuracy expected in the solutions. An adaptive mesh refinement technique provides better resolution in regions with strong flowfield gradients. Results for high-speed flow simulations are presented with the objective of assessing the implemented capability. Copyright © 2007 John Wiley & Sons, Ltd. [source] Evaluation of the biliary tract: The value of performing magnetic resonance cholangiopancreatography in conjunction with a 3-D spoiled gradient-echo gadolinium enhanced dynamic sequenceJOURNAL OF MEDICAL IMAGING AND RADIATION ONCOLOGY, Issue 4 2007SJ Kim Summary The 3-D gradient-echo (GRE) sequence allows thinner sections and better resolution of biliary obstruction. When the presence of biliary obstruction is identified using magnetic resonance cholangiopancreatography, the addition of the 3-D GRE sequence may be helpful for diagnosing biliary obstruction. By showing the changes in the bile duct wall, within the duct lumen and around the bile duct, this technique can be helpful for distinguishing benign from malignant stricture as well as a stone from an enhancing intraluminal mass. [source] Millipede phylogeny revisited in the light of the enigmatic order SiphoniulidaJOURNAL OF ZOOLOGICAL SYSTEMATICS AND EVOLUTIONARY RESEARCH, Issue 2 2003P. Sierwald Abstract The discovery of six specimens of the enigmatic order Siphoniulida, including for the first time males, prompted a modern re-analysis of current phylogenetic schemes for the class Diplopoda derived from traditional morphological and developmental characters. The data matrix was constructed and analysed using paup. The resulting phylogenetic hypotheses corroborated the longest standing, traditional classification, but also demonstrated clearly that more characters must be included to reach a better resolution. Recent alternative phylogenetic hypotheses and classifications are discussed in the light of the current analysis. The validity of a putative helminthomorph synapomorphy, the location of male gonopods on the 7th body ring, is discussed. Scanning electron microscopy corroborated morphological characters already described for the Siphoniulida: modified anterior legs, an apparently legless 3rd body ring, pyriform head, antennae with clavate setae, and absence of ozopores. The highly modified gonopods of the Siphoniulida are described for the first time; only the anterior legs of the 7th ring are modified into gonopods, the posterior legs of this ring are developed as normal walking legs. The gnathochilarium differs from the Colobognatha and consists of well-developed stipites with palps, elongated lingual plates with palps and a narrow central sclerite, most likely the mentum. Structures on the epiproct may possibly be spinnerets. Despite the discovery of adult males, the Siphoniulida are still considered Helminthomorpha incertae sedis . Zusammenfassung Die Entdeckung von 6 Exemplaren der ungewöhnlichen Ordnung Siphoniulida, einschließlich der ersten Männchen, erforderte eine Neu-Analyse der derzeitig diskutierten, morphologisch begründeten Klassifikationen der Klasse Diplopoda. Die erstellte Datenmatrize wurde in PAUP analysiert. Die sich ergebenden phylogenetischen Hypothesen bestätigen in wesentlichen die früheren, traditionellen Klassifikationsschemata; allerdings zeigt diese Analyse auch, dass die Merkmalsbasis erweitert werden muss, um eine höhere Auflösung zu erreichen. Neuere, abweichende phylogenetische Klassifikationen und Kladogramme werden unter Berücksichtigung der jetzigen Analyse verglichen. Die Gültigkeit einer der vorgeschlagenen Apomorphien für das Taxon Helminthomorpha, die Position der Gonopden am 7. Körperring, wird diskutiert. Rasterelektronenmikroskopische Untersuchungen bestätigten die morphologischen Merkmale, welche bereits für die Siphoniulida angegeben wurden: modifizierte Vorderbeine, scheinbar beinloser dritter Körperring, kleiner, zugespitzter Kopf, Antennen mit abgeflachten Borstenhaaren, fehlende Saftlöcher. Die männlichen Gonopoden werden hier zum ersten Male beschrieben: nur das vordere Beinpaar des 7. Ringes bildet die Gonopoden, das hintere Beinpaar desselben Ringes ist als normales Laufbeinpaar ausgebildet. Die Gonopoden sind stark modifizierte Strukturen. Das Gnathochilarium weicht deutlich von dem der Colobognathen ab, es besteht aus tastertragenden Stipites, langgestreckten Lingula Lamellae mit Tastern und einem zentralen, schmalen Sklerit, wahrscheinlich dem Mentum. Der Epiproct trägt vier Strukturen, die wahrscheinlich als Spinngriffel zu interpretieren sind. Trotz der Entdeckung von Männchen müssen die Siphoniulida weiterhin als Helminthomorpha incertae sedis angesehen werden. [source] Experimental determination of human peripheral nerve stimulation thresholds in a 3-axis planar gradient systemMAGNETIC RESONANCE IN MEDICINE, Issue 3 2009Rebecca E. Feldman Abstract In MRI, strong, rapidly switched gradient fields are desirable because they can be used to reduce imaging time, obtain images with better resolution, or improve image signal-to-noise ratios. Improvements in gradient strength can be made by either increasing the gradient amplifier strength or by enhancing gradient efficiency. Unfortunately, many MRI pulse sequences, in combination with high-performance amplifiers and existing gradient hardware, can cause peripheral nerve stimulation (PNS). This makes improvements in gradient amplifiers ineffective at increasing safely usable gradient strength. Customized gradient coils are one way to achieve significant improvements in gradient performance. One specific gradient configuration, a planar gradient system, promises improved gradient strength and switching time for cardiac imaging. The PNS thresholds for planar gradients were characterized through human stimulation experiments on all three gradient axes. The specialized gradient was shown to have significantly higher stimulation thresholds than traditional cylindrical designs (y-axis SRmin = 210 ± 18 mT/m/ms and ,Gmin = 133 ± 13 mT/m; x-axis SRmin = 222 ± 24 mT/m/ms and ,Gmin = 147 ± 17 mT/m; z-axis SRmin = 252 ± 26 mT/m/ms and ,Gmin = 218 ± 26 mT/m). This system could be operated at gradient strengths 2 to 3 times higher than cylindrical configurations without causing stimulation. Magn Reson Med, 2009. © 2009 Wiley-Liss, Inc. [source] HPLC determination of acidic d -amino acids and their N -methyl derivatives in biological tissuesBIOMEDICAL CHROMATOGRAPHY, Issue 6 2009Mara Tsesarskaia Abstract d -Aspartate (d -Asp) and N -methyl- d -aspartate (NMDA) occur in the neuroendocrine systems of vertebrates and invertebrates, where they play a role in hormone release and synthesis, neurotransmission, and memory and learning. N -methyl- d -glutamate (NMDG) has also been detected in marine bivalves. Several methods have been used to detect these amino acids, but they require pretreatment of tissue samples with o -phthaldialdehyde (OPA) to remove primary amino acids that interfere with the detection of NMDA and NMDG. We report here a one-step derivatization procedure with the chiral reagent N-, -(5-fluoro-2,4-dinitrophenyl)-(d or l)-valine amide, FDNP-Val-NH2, a close analog of Marfey's reagent but with better resolution and higher molar absorptivity. The diastereomers formed were separated by HPLC on an ODS-Hypersil column eluted with TFA/water,TFA/MeCN. UV absorption at 340 nm permitted detection levels as low as 5,10 pmol. d -Asp, NMDA and NMDG peaks were not obscured by other primary or secondary amino acids; hence pretreatment of tissues with OPA was not required. This method is highly reliable and fast (less than 40 min HPLC run). Using this method, we detected d -Asp, NMDA and NMDG in several biological tissues (octopus brain, optical lobe and bucchal mass; foot and mantle of the mollusk Scapharca broughtonii), confirming the results of other researchers. Copyright © 2009 John Wiley & Sons, Ltd. [source] Case study and application of process analytical technology (PAT) towards bioprocessing: Use of tryptophan fluorescence as at-line tool for making pooling decisions for process chromatographyBIOTECHNOLOGY PROGRESS, Issue 5 2009Anurag S. Rathore Abstract Process analytical technology (PAT) has been gaining momentum in the biopharmaceutical community due to the potential for continuous real time quality assurance resulting in improved operational control and compliance. Two imperatives for implementing any PAT tool are that "variability is managed by the process" and "product quality attributes can be accurately and reliably predicted over the design space established for materials used, process parameters, manufacturing, environmental, and other conditions." Recently, we have been examining the feasibility of applying different analytical tools to bioprocessing unit operations. We have previously demonstarted that commercially available online-high performance liquid chromatography and ultra performance liquid chromatography systems can be used for analysis that can facilitate real-time decisions for column pooling based on product quality attributes (Rathore et al., 2008a,b). In this article, we review an at-line tool that can be used for pooling of process chromatography columns. We have demonstrated that our tryptophan fluorescence method offers a feasible approach and meets the requirements of a PAT application. It is significantly faster than the alternative of fractionation, offline analysis followed by pooling. Although the method as presented here is not an online method, this technique may offer better resolution for certain applications and may be a more optimal approach as it is very conducive to implementation in a manufacturing environment. This technique is also amenable to be used as an online tool via front face fluorescence measurements done concurrently with product concentration determination by UV. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source] In vivo confocal microscopic evaluation of inflammatory changes in the ocular surfaceACTA OPHTHALMOLOGICA, Issue 2009A LABBE Purpose The ocular surface constitutes a complex physiopathological and anatomical entity assuring the barrier between the outside world and the fragile ocular structures. Ophthalmic instruments such as the slit lamp, which magnifies approximately 40 times, cannot provide details of the corneal structures at the cellular level. Methods In vivo confocal microscopy using the HRT Rostock Cornea module® (HRT / RCM) provides better resolution and therefore outlines distinctively in vivo inflammatory changes occurring in the ocular surface. Results In vivo confocal microscopy is capable of providing corneal, conjunctival and limbal cellular details in different ocular surface diseases such as dry eye, infectious keratitis, toxic keratitis, corneal intraepithelial neoplasia or vernal keratoconjunctivitis. Conclusion In correlation with ex vivo impression cytology analysis, in vivo confocal microscopy constitutes an interesting aid in the diagnosis and management of complex ocular surface conditions. [source] CEC separation of heterocyclic amines using methacrylate monolithic columnsELECTROPHORESIS, Issue 11 2007Elena Barceló-Barrachina Abstract Two methacrylate-based monolithic columns, one with a negatively charged group (sulfonic group) and another with a new monomer N,N -dimethylamino ethyl acrylate (DMAEA), were prepared and tested for the separation of basic compounds by CEC. This new monolithic stationary phase was prepared by the in situ polymerization of DMAEA with butyl methacrylate and ethylene dimethacrylate, using a ternary porogenic solvent consisting of water, 1-propanol and 1,4-butanediol. The performance of this column was evaluated by means of the analysis of a family of heterocyclic amines. Separation conditions such as pH, amount of organic modifier, ionic strength and elution mode (normal or counterdirectional flow) were studied. At the optimal running electrolyte composition, and using the counterdirectional mode, symmetrical electrochromatographic peaks were obtained, with the number of theoretical plates up to 30,000 and a good resolution between closely related peaks. The 2-acrylamido-2-methyl-1-propane-sulfonic acid column was used for CEC-MS, taking advantage of the compatibility of its elution mode (normal flow) with the MS coupling. [source] Development of off-line and on-line capillary electrophoresis methods for the screening and characterization of adenosine kinase inhibitors and substratesELECTROPHORESIS, Issue 12 2006Jamshed Iqbal Abstract Fast and convenient CE assays were developed for the screening of adenosine kinase,(AK) inhibitors and substrates. In the first method, the enzymatic reaction was performed in a test tube and the samples were subsequently injected into the capillary by pressure and detected by their UV absorbance at 260,nm. An MEKC method using borate buffer (pH,9.5) containing 100,mM SDS (method,A) was suitable for separating alternative substrates (nucleosides). For the CE determination of AMP formed as a product of the AK reaction, a phosphate buffer (pH,7.5 or 8.5) was used and a constant current (95,,A) was applied (method,B). The methods employing a fused-silica capillary and normal polarity mode provided good resolution of substrates and products of the enzymatic reaction and a short analysis time of less than 10,min. To further optimize and miniaturize the AK assays, the enzymatic reaction was performed directly in the capillary, prior to separation and quantitation of the product employing electrophoretically mediated microanalysis (EMMA, method,C). After hydrodynamic injection of a plug of reaction buffer (20,mM Tris-HCl, 0.2,mM MgCl2, pH,7.4), followed by a plug containing the enzyme, and subsequent injection of a plug of reaction buffer containing 1,mM,ATP, 100,,M adenosine, and 20,,M,UMP as an internal standard,(I.S.), as well as various concentrations of an inhibitor, the reaction was initiated by the application of 5,kV separation voltage (negative polarity) for 0.20,min to let the plugs interpenetrate. The voltage was turned off for 5,min (zero-potential amplification) and again turned on at a constant current of ,60,,A to elute the products within 7,min. The method employing a polyacrylamide-coated capillary of 20,cm effective length and reverse polarity mode provided good resolution of substrates and products. Dose,response curves and calculated Ki values for standard antagonists obtained by CE were in excellent agreement with data obtained by the standard radioactive assay. [source] Molecular phylogeny of Calyptratae (Diptera: Brachycera): the evolution of 18S and 16S ribosomal rDNAs in higher dipterans and their use in phylogenetic inferenceINSECT MOLECULAR BIOLOGY, Issue 5 2001X. Nirmala Abstract Sequences for nearly complete 18S rRNA and partial 16S rRNA genes were determined for sixteen species representing twelve calyptrate families. Two unique insertions are present in expansion regions of the 18S rDNA in nycteribiids. Alignments containing other dipteran rRNA genes provided good resolution at higher taxonomic level: monophyly of Calyptratae is well supported. While both 16S and 18S rDNA matrices produce unstable topologies within Calyptratae when analysed separately, their combination results in a tree with several robust and well supported nodes. Of three superfamilies recognized in recent classifications, the Hippoboscoidea is well supported by 16S rDNA and by combined matrices. The representatives of Muscoidea, Musca sp. and Antipoda sp., display a tendency to cluster within Oestroidea. The comparison of secondary structures of two variable regions indicates that Sarcophagidae are related to Calliphoridae rather than to Tachinidae. [source] Capillary Zone Electrophoresis and Micellar Electrokinetic Capillary Chromatography for Determining Water-Soluble Vitamins in Commercial Capsules and TabletsJOURNAL OF FOOD SCIENCE, Issue 1 2001S-C. Su ABSTRACT: A rapid method was developed for simultaneously determining thiamine, riboflavin, pyridoxine, nicotinamide, nicotinic acid, and ascorbic acid. It was tested on 15 samples. The peaks of all components were cleanly separated with good resolution by capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MECC). CZE was performed with 0.02 M borate buffer, and MECC was performed with 4% acetonitrile in 0.02 M borate/phosphate buffer containing 0.1 M sodium dodecyl sulfate. Average recoveries for all components were 80.3% to 103.7% with coefficients of variation being less than 5%. Thiamine, nicotinic acid, and pyridoxine contents were consistent with those labeled on the packages, but nicotinamide, riboflavin, and ascorbic acid contents of some samples were less. [source] Characterization of covalently inhibited extracellular lipase from Streptomyces rimosus by matrix-assisted laser desorption/ionization time-of-flight and matrix-assisted laser desorption/ionization quadrupole ion trap reflectron time-of-flight mass spectrometry: localization of the active site serine,JOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 12 2004Martin Zehl Abstract A chemical modification approach combined with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to identify the active site serine residue of an extracellular lipase from Streptomyces rimosus R6-554W. The lipase, purified from a high-level overexpressing strain, was covalently modified by incubation with 3,4-dichloroisocoumarin, a general mechanism-based serine protease inhibitor. MALDI time-of-flight (TOF) mass spectrometry was used to probe the nature of the intact inhibitor-modified lipase and to clarify the mechanism of lipase inhibition by 3,4-dichloroisocoumarin. The stoichiometry of the inhibition reaction revealed that specifically one molecule of inhibitor was bound to the lipase. The MALDI matrix 2,6-dihydroxyacetophenone facilitated the formation of highly abundant [M + 2H]2+ ions with good resolution compared to other matrices in a linear TOF instrument. This allowed the detection of two different inhibitor-modified lipase species. Exact localization of the modified amino acid residue was accomplished by tryptic digestion followed by low-energy collision-induced dissociation peptide sequencing of the detected 2-(carboxychloromethyl)benzoylated peptide by means of a MALDI quadrupole ion trap reflectron TOF instrument. The high sequence coverage obtained by this approach allowed the confirmation of the site specificity of the inhibition reaction and the unambiguous identification of the serine at position 10 as the nucleophilic amino acid residue in the active site of the enzyme. This result is in agreement with the previously obtained data from multiple sequence alignment of S. rimosus lipase with different esterases, which indicated that this enzyme exhibits a characteristic Gly-Asp-Ser-(Leu) motif located close to the N-terminus and is harboring the catalytically active serine residue. Therefore, this study experimentally proves the classification of the S. rimosus lipase as GDS(L) lipolytic enzyme. Copyright © 2004 John Wiley & Sons, Ltd. [source] HPLC/ESI-MSn and ESI-MS studies on the Aconitum alkaloids in three Chinese medicinal herbsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 17-18 2010Wenlong Liu Abstract Aconitum lipo-alkaloids (LDAs) peaks in the previous HPLC studies have shown overlapping behavior, making it impossible to identify the total retention behavior of LDAs. The structural diversity of LDAs in Radix Aconiti, Radix Aconiti Kusnezoffii and Radix Aconiti Lateralis Preparata makes it difficult to synthesize and quantify each of them. Therefore, this study has been devoted to develop a HPLC/ESI-MSn method with good resolution of Aconitum alkaloids especially LDAs and then set up a fast ESI-MS SIM method to quantify the total contents of specific skeleton LDAs in the three herbs via in-source CID and the capability of mass separation. As a result, the retention behavior of LDAs was summarized based on an optimized chromatographic separation; 32 alkaloids from the three herbs of Aconitum genus were separated and identified according to their fragmentation pathways by using HPLC/ESI-MSn. In addition, a fast quantitative analysis ESI-MS SIM method was successfully applied to the quantification of the total contents of specific skeleton LDAs in the three herbs. [source] Identification and determination of the saikosaponins in Radix bupleuri by accelerated solvent extraction combined with rapid-resolution LC-MSJOURNAL OF SEPARATION SCIENCE, JSS, Issue 13 2010Yun-Yun Yang Abstract A method based on accelerated solvent extraction combined with rapid-resolution LC,MS for efficient extraction, rapid separation, online identification and accurate determination of the saikosaponins (SSs) in Radix bupleuri (RB) was developed. The RB samples were extracted by accelerated solvent extraction using 70% aqueous ethanol v/v as solvent, at a temperature of 120°C and pressure of 100,bar, with 10,min of static extraction time and three extraction cycles. Rapid-resolution LC separation was performed by using a C18 column at gradient elution of water (containing 0.5% formic acid) and acetonitrile, and the major constituents were well separated within 20,min. A TOF-MS and an IT-MS were used for online identification of the major constituents, and 27 SSs were identified or tentatively identified. Five major bioactive SSs (SSa, SSc, SSd, 6,- O -acetyl-SSa and 6,- O -acetyl-SSd) with obvious peak areas and good resolution were chosen as benchmark substances, and a triple quadrupole MS operating in multiple-reaction monitoring mode was used for their quantitative analysis. A total of 16 RB samples from different regions of China were analyzed. The results indicated that the method was rapid, efficient, accurate and suitable for use in the quality control of RB. [source] Extended application of a chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid to the resolution of N -(substituted benzoyl)-,-amino acid amidesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 10 2006Guanghui Tan Abstract A chiral stationary phase (CSP 1) based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid was applied to the resolution of N -(substituted benzoyl)-,-amino acid amides and esters. N -(Substituted benzoyl)-,-amino acid amides were well resolved using a mixture of acetic acid-triethylamine-acetonitrile (0.01 : 0.05 : 100, v/v/v) as an optimum mobile phase while N -(substituted benzoyl)-,-amino acid esters were not resolved at all. In contrast, both N -(substituted benzoyl)-,-amino acid amides and esters were not resolved at all or resolved very poorly on another CSP (CSP 2), which lacks the two N,H hydrogens of the amide tethers of CSP 1. Among the substituents on the benzoyl group of analytes, the nitro group was the best for good resolution of analytes on CSP 1. From these results, the two N,H hydrogens of the amide tethers of CSP 1, the carbonyl oxygen of the amide group of analytes, and the nitro group on the benzoyl group of analytes were concluded to play significant roles in chiral recognition. In addition, various N -(3,5-dinitrobenzoyl)leucine amides with different lengths of N -alkylamide chains were resolved on CSP 1 and N -(3,5-dinitrobenzoyl)leucine N -propylamide was found to show the best chiral recognition in terms of the separation (, = 1.30) and the resolution factor (RS = 3.17). [source] Application of 31P NMR spectroscopy and chemical derivatization for metabolite profiling of lipophilic compounds in human serumMAGNETIC RESONANCE IN CHEMISTRY, Issue S1 2009M. Aruni DeSilva Abstract New methods for obtaining metabolic fingerprints of biological samples with improved resolution and sensitivity are highly sought for early disease detection, studies of human health and pathophysiology, and for better understanding systems biology. Considering the complexity of biological samples, interest in biochemical class selection through the use of chemoselective probes for improved resolution and quantitation is increasing. Considering the role of lipids in the pathogenesis of a number of diseases, in this study fingerprinting of lipid metabolites was achieved by 31P labeling using the derivatizing agent 2-chloro-4,4,5,5-tetramethyldioxaphospholane. Lipids containing hydroxyl, aldehyde and carboxyl groups were selectively tagged with 31P and then detected with good resolution using 31P NMR by exploiting the 100% natural abundance and wide chemical shift range of 31P. After standardizing the reaction conditions using representative compounds, the derivatization approach was used to profile lipids in human serum. The results show that the 31P derivatization approach is simple, reproducible and highly quantitative, and has the potential to profile a number of important lipids in complex biological samples. Copyright © 2009 John Wiley & Sons, Ltd. [source] MMIC's characterization by very near-field techniqueMICROWAVE AND OPTICAL TECHNOLOGY LETTERS, Issue 3 2004L. Nativel Abstract This paper shows a method to characterize microwave circuits using a near-field scanning microscope. Applied on various samples, it shows good resolution and weak disturbance for ICs operating with very common microwave components. Here, it is applied in an industrial surrounding to characterize the Bluetooth CMOS power amplifier. © 2004 Wiley Periodicals, Inc. Microwave Opt Technol Lett 41: 209,213, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.20096 [source] Combination of immunohistochemistry and laser ablation ICP mass spectrometry for imaging of cancer biomarkersPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 18 2008Jaume Seuma Abstract Laser ablation (LA) ICP-MS has been developed as a new tool for imaging of cancer biomarkers in tissue sections. The distribution of two breast cancer-associated proteins, MUC-1 and HER2 was studied based on multiple line rastering of tissue sections and measurement of relevant Au/Ag tagged antibodies bound to the tissue. Comparisons with optical microscopy indicated extremely high sensitivity for the LA technique and sufficiently good resolution to permit fine scale feature mapping at the cellular level. Application to the quantitative assessment of HER2 expression in tissue microarrays was demonstrated. [source] Determination of ibudilast in human serum by high-performance liquid chromatography for pharmacokinetic studyBIOMEDICAL CHROMATOGRAPHY, Issue 3 2010Hwa Yoon Abstract A simple, accurate, precise and cost effective reversed-phase HPLC method was developed to determine the concentration of ibudilast in human serum. Ibudilast and an internal standard, butyl 4-hydroxybenzoate, were extracted by liquid,liquid extraction with methyl tert -butyl ether. HPLC analysis was carried out under the following conditions: a Luna C18(2) 5,,m column, a mobile phase of acetonitrile,0.02% phosphoric acid (50,:,50, v/v, adjusted to pH 6.0 with triethylamine) and a UV detector at 319,nm. The chromatograms showed good resolution and sensitivity as well as no interference from the human serum. The calibration curves were linear over the concentration range, 1,100,ng/mL, for serum with correlation coefficients >0.999. The intra- and inter-day assay precision as well as the accuracy fulfilled the international requirements. The mean absolute recovery for human serum was 101.7 ± 6.1%. The lower limit of quantitation in human serum was 1,ng/mL, which is sensitive enough for pharmacokinetic studies. Stability studies revealed that ibudilast in human serum was stable during storage as well as during the assay procedure. This method was applied successfully to an examination of the pharmacokinetics of ibudilast in human subjects following a single oral dose of an ibudilast (10,mg) capsule. Copyright © 2009 John Wiley & Sons, Ltd. [source] Synthesis, X-ray Single-Crystal Structure Determination, and Magnetic Properties of [Rb(benzo[18]crown-6)]+ Salts Containing Well-Ordered Fulleride Trianions C603,,CHEMISTRY - A EUROPEAN JOURNAL, Issue 13 2009Bele Boeddinghaus Dipl.-Chem. Abstract Finally, a structure with well-resolved C603,ions and S=1/2 spin system: The two novel title compounds have clearly S=1/2 and not S=3/2 electronic states, as expected for the occupation of the triply degenerate LUMO of C60. These structures with well-ordered fullerene trianions show that the expected Jahn,Teller distortion is not observable in X-ray diffraction experiments. Crystals of the composition [Rb(benzo[18]crown-6)]3[,3 -C60](C7H8)(C3H7NO)4.5 (1) and [Rb(benzo[18]crown-6)]3[C60](C3H7NO)5.33(C4H8O)1.66 (2) were synthesized by reduction of C60 with elemental rubidium in a mixture of dimethylformamide/tetrahydrofuran in the presence of benzo[18]crown-6. X-ray single-crystal structure determinations reveal well-ordered C60 trianions in both compounds. In contrast to previously reported structures of compounds containing C603, ions the present structure refinements allow the description of the buckyballs with good resolution (R1/wR2 for I>2,(I) (all data) are 0.030/0.069 (0.051/0.071) and 0.042/0.101 (0.076/0.108) for 1 and 2, respectively). In 1 the C603, unit coordinates in an ,3 -fashion to one Rb atom, whose coordination sphere is completed by one benzo[18]crown-6 and one dimethylformamide molecule. In 2 no coordination of the alkali metal atom to the C60 trianion is observed. Magnetic measurements for 1 reveal a magnetic moment of 1.74,,B, as expected for an isolated S=1/2 spin system. The high-quality data of the present studies allowed a comparison of structural details of C603, ions with other C60n, units (n=0, 2, 3) and a discussion of the structural Jahn,Teller distortions. [source] Breaking good resolutions with ARP/wARPJOURNAL OF SYNCHROTRON RADIATION, Issue 1 2004Richard J. Morris New procedures are outlined that enable ARP/wARP to automatically build protein models with diffraction data extending to about 2.5 Å. An overview of ongoing research is given and possible future advances are discussed. [source] | |||||||||||||||||||