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Various Matrices (various + matrix)
Selected AbstractsElectronic gel protein transfer and identification using matrix-assisted laser desorption/ionization-mass spectrometryELECTROPHORESIS, Issue 9 2004Jonathan W. Cooper Abstract An electronic protein transfer technique is described for achieving the rapid and efficient recovery of sodium dodecyl sulfate (SDS)-protein complexes from polyacrylamide gels. This process involves the use of small-dimension capillaries in physical contact with a resolved protein band within the polyacrylamide gel, providing a large potential drop and high electric field strength at the capillary/gel interface. Several factors controlling the electronic protein transfer, including the applied electric field strength, the electrophoresis buffer concentration, and the capillary dimension, are studied to further enhance the use of field-amplification for sample stacking of extracted SDS-protein complexes. As a result of sample stacking, the extracted proteins from a 50 ng gel loading are present in a narrow (,80 nL) and highly concentrated (0.46 mg/mL or 3.3×10,5 M for cytochrome c) solution plug. Three model proteins with molecular mass ranging from 14 kDa (cytochrome c) to 116 kDa (,-galactosidase) are stained by Coomassie blue and electrophoretically extracted from gels with protein loadings as low as 50 ng. The capillary format of the electronic protein transfer technique allows direct deposition of extracted proteins onto a matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) target. Various matrices and solvent compositions are evaluated for the analysis of extracted and concentrated SDS-protein complexes using MALDI-MS. The electronic protein transfer technique, when operated under optimized conditions, is demonstrated for the effective (>70% recovery), speedy (less than 5 min), and sensitive MS identification of gel resolved proteins (as low as 50 ng). [source] Imaging mass spectrometry for examining localization of polymeric composition in matrix-assisted laser desorption/ionization samplesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 5 2009Steffen M. Weidner The localization of polymeric composition in samples prepared for matrix-assisted laser desorption/ionization (MALDI) analysis has been investigated by imaging mass spectrometry. Various matrices and solvents were used for sample spot preparation of a polybutyleneglycol (PBG 1000). It was shown that in visibly homogeneous spots, prepared using the ,dried droplet' method, separation between matrix and polymer takes place. Moreover, using , -cyano-4-hydroxycinnamic acid (CCA) as matrix and methanol as solvent molecular mass separation of the polymer homologues in the spots was detectable. In contrast to manually spotted samples, dry spray deposition results in homogeneous layers showing no separation effects. Copyright © 2009 John Wiley & Sons, Ltd. [source] Determination of biogenic amines in HeLa cell lysate by 6-oxy-(N -succinimidyl acetate)-9-(2',methoxycarbonyl) fluorescein and micellar electrokinetic capillary chromatography with laser-induced fluorescence detectionELECTROPHORESIS, Issue 4 2006Liwei Cao Abstract An MEKC-LIF method using 6-oxy-(N -succinimidyl acetate)-9-(2'-methoxy-carbonyl) fluorescein (SAMF) newly synthesized in our lab as a labeling reagent for the separation and determination of eight typical biogenic amines was proposed. After careful study of the derivatization condition such as pH value, reagent concentration, temperature, and reaction time, derivatization reaction was accomplished as quickly as 10,min with stable yield. Optimal separation of SAMF-labeled amines was achieved with a running buffer (pH,9.3) containing 30,mM boric acid, 25,mM SDS, and 20%,v/v ACN. The proposed method allowed biogenic amines to be determined with LODs as low as 0.25,2.5,nmol/L and RSD values from 0.4 to 4.5%. The present method has been successfully used to monitor biogenic amines in HeLa cells and fish samples. This study exploits the potential of MEKC-LIF with SAMF labeling as a tool for monitoring biogenic amines involved in complex physiological and behavioral processes in various matrices. [source] THE ADDITIVE GENETIC VARIANCE AFTER BOTTLENECKS IS AFFECTED BY THE NUMBER OF LOCI INVOLVED IN EPISTATIC INTERACTIONSEVOLUTION, Issue 4 2003Yamama Naciri-Graven Abstract We investigated the role of the number of loci coding for a neutral trait on the release of additive variance for this trait after population bottlenecks. Different bottleneck sizes and durations were tested for various matrices of genotypic values, with initial conditions covering the allele frequency space. We used three different types of matrices. First, we extended Cheverud and Routman's model by defining matrices of "pure" epistasis for three and four independent loci; second, we used genotypic values drawn randomly from uniform, normal, and exponential distributions; and third we used two models of simple metabolic pathways leading to physiological epistasis. For all these matrices of genotypic values except the dominant metabolic pathway, we find that, as the number of loci increases from two to three and four, an increase in the release of additive variance is occurring. The amount of additive variance released for a given set of genotypic values is a function of the inbreeding coefficient, independently of the size and duration of the bottleneck. The level of inbreeding necessary to achieve maximum release in additive variance increases with the number of loci. We find that additive-by-additive epistasis is the type of epistasis most easily converted into additive variance. For a wide range of models, our results show that epistasis, rather than dominance, plays a significant role in the increase of additive variance following bottlenecks. [source] Analysis of flavor and perfume using an internally cooled coated fiber deviceJOURNAL OF SEPARATION SCIENCE, JSS, Issue 7 2007Yong Chen Abstract A miniaturized internally cooled coated fiber device was applied for the analysis of flavors and fragrances from various matrices. Its integration with a CTC CombiPAL autosampler enabled high throughput for the analysis of analytes in complex matrices that required simultaneous heating of the matrices and cooling of the fiber coating to achieve high extraction efficiency. It was found that up to ten times increase of extraction efficiencies was observed when the device was used to extract flavor compounds in water, even when limited sample temperatures were used to preserve the integrity of target compounds. The extraction of the flavor compounds in water with the device was reproducible, with RSD not larger than 15%. The lower limits of the linear ranges were in the low ppb range, which was about one order of magnitude smaller than those obtained with the commercialized 100 ,m PDMS fibers. Exhaustive extraction of some perfume ingredients from a complex matrix (shampoo) was realized. All achieved recoveries were not less than 80%. The repeatability of the extraction of the perfume compounds from shampoo was better than 10%. The linear ranges were about 1,3000 ,g/g, and the LOD was about 0.2,1 ,g/g. The automated internally cooled coated fiber device was demonstrated to be a powerful sample preparation tool in flavor and fragrance analysis. [source] Matrix effects in quantitative pesticide analysis using liquid chromatography,mass spectrometryMASS SPECTROMETRY REVIEWS, Issue 6 2006W.M.A. Niessen Abstract Combined liquid chromatography,mass spectrometry using electrospray or atmospheric-pressure chemical ionization has become an important tool in the quantitative analysis of pesticide residues in various matrices in relation to environmental analysis, food safety, and biological exposure monitoring. One of the major problems in the quantitative analysis using LC,MS is that compound and matrix-dependent response suppression or enhancement may occur, the so-called matrix effect. This article reviews issues related to matrix effects, focusing on quantitative pesticide analysis, but also paying attention to expertise with respect to matrix effects acquired in other application areas of LC,MS, especially quantitative bioanalysis in the course of drug development. © 2006 Wiley Periodicals, Inc. [source] Brominated flame retardants in US foodMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 2 2008Arnold Schecter Abstract We and others recently began studying brominated flame retardant levels in various matrices in the US including human milk and other food. This paper reviews the food studies. In our studies, ten to thirteen polybrominated diphenyl ether (PBDE) congeners were measured, usually including BDE 209. All US women's milk samples were contaminated with PBDEs from 6 to 419 ng/g, lipid, orders of magnitude higher than levels reported in European studies, and are the highest reported worldwide. We compared our market basket studies of meat, fish and dairy products with other US food studies of meat and fish. US studies showed somewhat higher levels of PBDEs than reported elsewhere. Fish were most highly contaminated (median 616 pg/g), then meat (median190 pg/g) and dairy products (median 32.2 pg/g). However, unlike some European countries where fish predominates, dietary intake of PBDEs in the US is mostly from meat, then fish and then dairy products. Broiling can decrease the amount of PBDEs per serving. We also measured levels of hexabromocyclododecane (HBCD), another brominated flame retardant, in human milk. The levels are lower than PBDEs, 0.16,1.2 ng/g, similar to European levels, unlike PBDEs where US levels are much higher than European levels. [source] Off-line liquid chromatography-MALDI by with various matrices and tandem mass spectrometry for analysis of glycated human serum albumin tryptic peptidesMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 4 2007Annunziata Lapolla Abstract Advanced glycation end-product (AGE)/peptides, arising from in vivo digestion of glycated proteins, are biologically important compounds, due to their reactivity against circulating and tissue proteins. For information on their possible structure, in vitro glycation of HSA and its further enzymatic digestion were performed. The resulting digestion product mixture was analysed directly by MALDI MS with various matrices [2,5-dihydroxy benzoic acid (DHB) and ,-cyano-4-hydroxy cinnamic acid (CHCA)]. Alternatively, offline microbore LC prior to MALDI analysis was used, and showed that 63% of the free amino groups prone to glycation are modified, indicating the contemporary presence of unglycated peptides. This result proves that, regardless of the high glucose concentration employed for HSA incubation, glycation does not go to completion. Further studies showed that the collisionally activated decomposition of singly charged glycated peptides leads to specific fragmentation pathways, all related to the condensed glucose molecule. These unique product ions can be used as effective markers to establish the presence of a glucose molecule within a peptide ion. [source] Rapid gas chromatography/mass spectrometry quinine determination in plasma after automated solid-phase extractionRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2006Richard Damien The combined use of an automatic solid-phase extraction (SPE) apparatus with Oasis MCX cartridges and gas chromatography/mass spectrometry (GC/MS) to rapidly quantify quinine in biological samples with cyproheptadine as the internal standard is described. The selected ion monitoring mode, with the quantification ions m/z 136 and 287 (qualifier ions: m/z 261, 381 and 215, 96), allows the estimation of quinine levels, respectively. Separation was completed within 12.7,min. Excellent linearity was found up to 10 000,µg/L of plasma. The limit of detection (LOD) was 12.2,µg/L and the limit of quantification (LOQ) was 40.6,µg/L. High reproducibility (intra-assay CV range 1.9,4.3%, inter-assay CV range 2.2,11.3%) and accuracy values (intra-assay range 83.2,103.7%, inter-assay range 86.8,103.7%) were obtained. Recoveries were concentration-independent (97.2% and 89.8% for 4000 and 10 000,µg/L, respectively). This sensitive, simple assay for quinine in various matrices meets the current requirements for bioanalytical assays and may be used to monitor quinine levels in patients developing severe malaria with acute renal failure during hemofiltration. The optimal quinine dose in this situation is not really established and to improve clinical care, quinine concentrations might be explored to improve efficacy and minimise potential toxicity. Copyright © 2006 John Wiley & Sons, Ltd. [source] Matrix-assisted laser desorption/ionization imaging mass spectrometry for direct measurement of clozapine in rat brain tissueRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 6 2006Yunsheng Hsieh Matrix-assisted laser desorption/ionization hyphenated with quadrupole time-of-flight (QTOF) mass spectrometry (MS) has been used to directly determine the distribution of pharmaceuticals in rat brain tissue slices which might unravel their disposition for new drug development. Clozapine, an antipsychotic drug, and norclozapine were used as model compounds to investigate fundamental parameters such as matrix and solvent effects and irradiance dependence on MALDI intensity but also to address the issues with direct tissue imaging MS technique such as (1) uniform coating by the matrix, (2) linearity of MALDI signals, and (3) redistribution of surface analytes. The tissue sections were coated with various matrices on MALDI plates by airspray deposition prior to MS detection. MALDI signals of analytes were detected by monitoring the dissociation of the individual protonated molecules to their predominant MS/MS product ions. The matrices were chosen for tissue applications based on their ability to form a homogeneous coating of dense crystals and to yield greater sensitivity. Images revealing the spatial localization in tissue sections using MALDI-QTOF following a direct infusion of 3H-clozapine into rat brain were found to be in good correlation with those using a radioautographic approach. The density of clozapine and its major metabolites from whole brain homogenates was further confirmed using fast high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) procedures. Copyright © 2006 John Wiley & Sons, Ltd. [source] Kinetic monitoring of trisubstituted organotins in soil after sewage sludge applicationAPPLIED ORGANOMETALLIC CHEMISTRY, Issue 9 2008S. Dubascoux Abstract Organotin compounds (OTC) are widely used for their biocidal effects in various agricultural or industrial activities, leading to their environmental presence. Among the organotin species, tributyltin (TBT) and triphenyltin (TPhT) are the most used and are generally considered the most toxic. So it is important to understand their behaviour in soils and obtain data about their persistence and phytoavailability. Many works deal with OTC speciation in various matrices, but few are concerned with OTC degradation in soil. The present study focuses on kinetic monitoring of TBT and TPhT in an agricultural soil. These compounds were introduced into the soil by the way of spiked sewage sludge, simulating agricultural practice and diffuse contamination. The influence of time and initial OTC concentration on the species preservation was evaluated. TBT concentration was shown to have a positive effect on TBT preservation. Corresponding half-lives were calculated. They were 6 ± 1 days and over 39 days for TPhT and TBT, respectively. Degradation compounds, mono- and dibutyltin, and mono- and diphenyltin, were produced by both direct and successive dealkyl and dearylation processes. Copyright © 2008 John Wiley & Sons, Ltd. [source] Chromatographic determination of herbicide residues in various matricesBIOMEDICAL CHROMATOGRAPHY, Issue 6 2004Tibor Cserháti Abstract The newest results in the use of various extraction techniques and chromatographic methods such as gas,liquid and high-performance liquid chromatography used for the assessment of herbicide residues in various matrices have been compiled and critically evaluated. Practical employments in water and soil research, environmental protection, clinical and food chemistry are presented. Copyright © 2004 John Wiley & Sons, Ltd. [source] Noggin maintains pluripotency of human embryonic stem cells grown on MatrigelCELL PROLIFERATION, Issue 4 2009G. Chaturvedi Objective:, Spontaneous differentiation of human embryonic stem cell (hESC) cultures is a major concern in stem cell research. Physical removal of differentiated areas in a stem cell colony is the current approach used to keep the cultures in a pluripotent state for a prolonged period of time. All hESCs available for research require unidentified soluble factors secreted from feeder layers to maintain the undifferentiated state and pluripotency. Under experimental conditions, stem cells are grown on various matrices, the most commonly used being Matrigel. Materials and Methods:, We propose an alternative method to prevent spontaneous differentiation of hESCs grown on Matrigel that uses low amounts of recombinant noggin. We make use of the porosity of Matrigel to serve as a matrix that traps noggin and gradually releases it into the culture to antagonize bone morphogenetic proteins (BMP). BMPs are known to initiate differentiation of hESCs and are either present in the conditioned medium or are secreted by hESCs themselves. Results:, hESCs grown on Matrigel supplemented with noggin in conditioned medium from feeder layers (irradiated mouse embryonic fibroblasts) retained both normal karyotype and markers of hESC pluripotency for 14 days. In addition, these cultures were found to have increased cell proliferation of stem cells as compared to hESCs grown on Matrigel alone. Conclusion:, Noggin can be utilized for short term prevention of spontaneous differentiation of stem cells grown on Matrigel. [source] Toughened Oxide Composites Based on Porous Alumina-Platelet InterphasesJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 4 2001Sang-Jin Lee A novel mechanism for debonding at a weak interphase in an all-oxide composite is introduced. This methodology involves the use of alumina platelets that have a diameter of 10,15 or 5,10 ,m and a thickness of 1 ,m. The platelets induce constrained sintering of the ceramic powder, which results in permanent porosity. For room-temperature properties, only minor additions (0,3 vol%) of matrix powder yield sufficiently weak debonding interphases. The platelets lie in random, three-dimensional orientations and provide a debonding mechanism that is independent of temperature, in chemically compatible matrixes. Laminated composites with two types of matrixes,mullite and alumina,have been fabricated with modified fibrous monoliths of alumina in a triple-layer "core/interphase/matrix" arrangement. In the laminated systems, the intimate mixing of strong versus tough microstructures has been tailored by alternating various matrix:interphase thickness ratios. Preliminary load,displacement curves clearly demonstrate characteristics of "graceful failure" and notable improvements in the work of fracture. Scanning electron microscopic observation of the crack paths confirms the viability of platelets for producing permanently porous, debondable interphases at elevated temperatures in air. [source] | |||||||||||||