Showing papers in "Analytical Chemistry in 2006"
TL;DR: An LC/MS-based data analysis approach, XCMS, which incorporates novel nonlinear retention time alignment, matched filtration, peak detection, and peak matching, and is demonstrated using data sets from a previously reported enzyme knockout study and a large-scale study of plasma samples.
Abstract: Metabolite profiling in biomarker discovery, enzyme substrate assignment, drug activity/specificity determination, and basic metabolic research requires new data preprocessing approaches to correlate specific metabolites to their biological origin. Here we introduce an LC/MS-based data analysis approach, XCMS, which incorporates novel nonlinear retention time alignment, matched filtration, peak detection, and peak matching. Without using internal standards, the method dynamically identifies hundreds of endogenous metabolites for use as standards, calculating a nonlinear retention time correction profile for each sample. Following retention time correction, the relative metabolite ion intensities are directly compared to identify changes in specific endogenous metabolites, such as potential biomarkers. The software is demonstrated using data sets from a previously reported enzyme knockout study and a large-scale study of plasma samples. XCMS is freely available under an open-source license at http://metlin...
3,963 citations
TL;DR: The development of a new high-resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS) is reported, which allows the direct separation of most ions from inorganic and organic species at the same nominal m/z, and the quantification of several types of organic fragments.
Abstract: The development of a new high-resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS) is reported. The high-resolution capabilities of this instrument allow the direct separation of most ions from inorganic and organic species at the same nominal m/z, the quantification of several types of organic fragments (CxHy, CxHyOz, CxHyNp, CxHyOzNp), and the direct identification of organic nitrogen and organosulfur content. This real-time instrument is field-deployable, and its high time resolution (0.5 Hz has been demonstrated) makes it well-suited for studies in which time resolution is critical, such as aircraft studies. The instrument has two ion optical modes: a single-reflection configuration offers higher sensitivity and lower resolving power (up to ∼2100 at m/z 200), and a two-reflectron configuration yields higher resolving power (up to ∼4300 at m/z 200) with lower sensitivity. The instrument also allows the determination of the size distributions of all ions. One-minute detection limits for subm...
1,818 citations
TL;DR: The probabilistic quotient normalization is introduced in this work, which is based on the calculation of a most probable dilution factor by looking at the distribution of the quotients of the amplitudes of a test spectrum by those of a reference spectrum.
Abstract: For the analysis of the spectra of complex biofluids, preprocessing methods play a crucial role in rendering the subsequent data analyses more robust and accurate. Normalization is a preprocessing method, which accounts for different dilutions of samples by scaling the spectra to the same virtual overall concentration. In the field of 1H NMR metabonomics integral normalization, which scales spectra to the same total integral, is the de facto standard. In this work, it is shown that integral normalization is a suboptimal method for normalizing spectra from metabonomic studies. Especially strong metabonomic changes, evident as massive amounts of single metabolites in samples, significantly hamper the integral normalization resulting in incorrectly scaled spectra. The probabilistic quotient normalization is introduced in this work. This method is based on the calculation of a most probable dilution factor by looking at the distribution of the quotients of the amplitudes of a test spectrum by those of a refer...
1,454 citations
TL;DR: N nanoparticles up to 250 nm in diameter were found have approximately 2 orders of magnitude higher DNA loading than smaller (13-30 nm) nanoparticles, a consequence of their larger surface area.
Abstract: We have investigated the variables that influence DNA coverage on gold nanoparticles. The effects of salt concentration, spacer composition, nanoparticle size, and degree of sonication have been evaluated. Maximum loading was obtained by salt aging the nanoparticles to ∼0.7 M NaCl in the presence of DNA containing a poly(ethylene glycol) spacer. In addition, DNA loading was substantially increased by sonicating the nanoparticles during the surface loading process. Last, nanoparticles up to 250 nm in diameter were found have ∼2 orders of magnitude higher DNA loading than smaller (13−30 nm) nanoparticles, a consequence of their larger surface area. Stable large particles are attractive for a variety of biodiagnostic assays.
1,073 citations
TL;DR: In this paper, the authors present a review of the book.http://www.reviewreviews.com/reviews/book-reviews-of-the-book
Abstract: Review
941 citations
TL;DR: The technique is validated against a traditional "spectral binning" analysis on the basis of sensitivity to water suppression, relaxation effects, and NMR spectral acquisition times using PCA pattern recognition analysis.
Abstract: Extracting meaningful information from complex spectroscopic data of metabolite mixtures is an area of active research in the emerging field of “metabolomics”, which combines metabolism, spectroscopy, and multivariate statistical analysis (pattern recognition) methods. Chemometric analysis and comparison of 1H NMR1 spectra is commonly hampered by intersample peak position and line width variation due to matrix effects (pH, ionic strength, etc.). Here a novel method for mixture analysis is presented, defined as “targeted profiling”. Individual NMR resonances of interest are mathematically modeled from pure compound spectra. This database is then interrogated to identify and quantify metabolites in complex spectra of mixtures, such as biofluids. The technique is validated against a traditional “spectral binning” analysis on the basis of sensitivity to water suppression (presaturation, NOESY-presaturation, WET, and CPMG), relaxation effects, and NMR spectral acquisition times (3, 4, 5, and 6 s/scan) using PC...
872 citations
TL;DR: Rapid, automated data-dependent capabilities enable real-time acquisition of up to three high-mass accuracy MS/MS spectra per second.
Abstract: Design and performance of a novel hybrid mass spectrometer is described. It couples a linear ion trap mass spectrometer to an orbitrap mass analyzer via an rf-only trapping quadrupole with a curved axis. The latter injects pulsed ion beams into a rapidly changing electric field in the orbitrap wherein they are trapped at high kinetic energies around an inner electrode. Image current detection is subsequently performed after a stable electrostatic field is achieved. Fourier transformation of the acquired transient allows wide mass range detection with high resolving power, mass accuracy, and dynamic range. The entire instrument operates in LC/MS mode (1 spectrum/s) with nominal mass resolving power of 60 000 and uses automatic gain control to provide high-accuracy mass measurements, within 2 ppm using internal standards and within 5 ppm with external calibration. The maximum resolving power exceeds 100 000 (fwhm). Rapid, automated data-dependent capabilities enable real-time acquisition of up to three high...
762 citations
748 citations
TL;DR: In this paper, it is recommended that δ13C values of both organic and inorganic materials be measured and expressed relative to VPDB (Vienna Peedee belemnite) on a scale normalized by assigning consensus values of −46.6
Abstract: Consistency of δ13C measurements can be improved 39−47% by anchoring the δ13C scale with two isotopic reference materials differing substantially in 13C/12C. It is recommended that δ13C values of both organic and inorganic materials be measured and expressed relative to VPDB (Vienna Peedee belemnite) on a scale normalized by assigning consensus values of −46.6‰ to L-SVEC lithium carbonate and +1.95‰ to NBS 19 calcium carbonate. Uncertainties of other reference material values on this scale are improved by factors up to two or more, and the values of some have been notably shifted: the δ13C of NBS 22 oil is −30.03‰.
726 citations
TL;DR: A pragmatic approach combining several well-established statistical methods was developed for processing this large data set in order to detect small differences in metabolic profiles in combination with a large biological variation and proved to be applicable for dealing with large-scale human metabolomics studies.
Abstract: A large metabolomics study was performed on 600 plasma samples taken at four time points before and after a single intake of a high fat test meal by obese and lean subjects. All samples were analyzed by a liquid chromatography-mass spectrometry (LC-MS) lipidomic method for metabolic profiling. A pragmatic approach combining several well-established statistical methods was developed for processing this large data set in order to detect small differences in metabolic profiles in combination with a large biological variation. Such metabolomics studies require a careful analytical and statistical protocol. The strategy included data preprocessing, data analysis, and validation of statistical models. After several data preprocessing steps, partial least-squares discriminant analysis (PLS-DA) was used for finding biomarkers. To validate the found biomarkers statistically, the PLS-DA models were validated by means of a permutation test, biomarker models, and noninformative models. Univariate plots of potential biomarkers were used to obtain insight in up- or downregulation. The strategy proposed proved to be applicable for dealing with large-scale human metabolomics studies. © 2006 American Chemical Society.
724 citations
TL;DR: This review of ionic liquids research highlights early work involving ILs and discusses the most recent advances in separations, mass spectrometry, spectroscopy, and electroanalytical chemistry.
Abstract: The role of ionic liquids (ILs) in analytical chemistry is increasing substantially every year. A decade ago there were but a handful of papers in this area of research that were considered curiosities at best. Today, those publications are recognized as seminal articles that gave rise to one of the most rapidly expanding areas of research in chemical analysis. In this review, we briefly highlight early work involving ILs and discuss the most recent advances in separations, mass spectrometry, spectroscopy, and electroanalytical chemistry. Many of the most important advances in these fields depend on the development of new, often unique ILs and multifunctional ILs. A better understanding of the chemical and physical properties of ILs is also essential.
TL;DR: It is concluded that data preprocessing is context dependent and there can be no single method for general use and care must be exercised in calculation of correlation coefficients for data sets where normalization to a constant sum is used.
Abstract: Considerable confusion appears to exist in the metabonomics literature as to the real need for, and the role of, preprocessing the acquired spectroscopic data. A number of studies have presented various data manipulation approaches, some suggesting an optimum method. In metabonomics, data are usually presented as a table where each row relates to a given sample or analytical experiment and each column corresponds to a single measurement in that experiment, typically individual spectral peak intensities or metabolite concentrations. Here we suggest definitions for and discuss the operations usually termed normalization (a table row operation) and scaling (a table column operation) and demonstrate their need in 1H NMR spectroscopic data sets derived from urine. The problems associated with "binned" data (i.e., values integrated over discrete spectral regions) are also discussed, and the particular biological context problems of analytical data on urine are highlighted. It is shown that care must be exercised in calculation of correlation coefficients for data sets where normalization to a constant sum is used. Analogous considerations will be needed for other biofluids, other analytical approaches (e.g., HPLC-MS), and indeed for other "omics" techniques (i.e., transcriptomics or proteomics) and for integrated studies with "fused" data sets. It is concluded that data preprocessing is context dependent and there can be no single method for general use.
TL;DR: IMS analysis of tissues from 8 mg/kg olanzapine dosed rats revealed temporal distribution of the drug and metabolites that correlate to previous quantitative whole- body autoradiography studies, and Whole-body MALDI IMS is further extended to detecting proteins from organs present in a whole-body sagittal tissue section.
Abstract: Imaging mass spectrometry (IMS) that utilizes matrix-assisted laser desorption/ionization (MALDI) technology can provide a molecular ex vivo view of resected organs or whole-body sections from an animal, making possible the label-free tracking of both endogenous and exogenous compounds with spatial resolution and molecular specificity. Drug distribution and, for the first time, individual metabolite distributions within whole-body tissue sections can be detected simultaneously at various time points following drug administration. IMS analysis of tissues from 8 mg/kg olanzapine dosed rats revealed temporal distribution of the drug and metabolites that correlate to previous quantitative whole-body autoradiography studies. Whole-body MALDI IMS is further extended to detecting proteins from organs present in a whole-body sagittal tissue section. This technology will significantly help advance the analysis of novel therapeutics and may provide deeper insight into therapeutic and toxicological processes, reveal...
TL;DR: This work presents the first demonstration of the utility of zirconium dioxide microtips for phosphopeptide isolation prior to mass spectrometric analysis, and shows that these microtips display similar overall performance as TiO2 microtips.
Abstract: Due to the dynamic nature and low stoichiometry of protein phosphorylation, enrichment of phosphorylated peptides from proteolytic mixtures is often necessary prior to their characterization by mass spectrometry. Several phosphopeptide isolation strategies have been presented in the literature, including immobilized metal ion affinity chromatography. However, that technique suffers from poor selectivity and reproducibility. Recently, titanium dioxide-based columns have been successfully employed for phosphopeptide enrichment by several research groups. Here, we present, to our knowledge, the first demonstration of the utility of zirconium dioxide microtips for phosphopeptide isolation prior to mass spectrometric analysis. These microtips display similar overall performance as TiO2 microtips. However, more selective isolation of singly phosphorylated peptides was observed with ZrO2 compared to TiO2 whereas TiO2 preferentially enriched multiply phosphorylated peptides. Thus, these two chromatographic materi...
TL;DR: This technology was applied to determine novel single-cell enzyme kinetics for three different cell types (HeLa, 293T, Jurkat) and predicted this varied response was due to variation in the concentration of carboxylesterase between cell types.
Abstract: High-quality single-cell data are required for a quantitative systems biology description of cellular function. However, data of this type are difficult and time-consuming to collect using traditional techniques. We present a robust and simple microfluidic method for trapping single cells in large arrays to address this problem. Ordered single-cell isolation arrays allow for high-density microscopic analysis with simplified image processing. Moreover, for fluorescent assays, on-chip sample preparation (e.g., fluorescent labeling, washing) can be performed, as opposed to manual intensive operations of incubation, centrifugation, and resuspension in previous techniques-saving time and reagents. This technology was applied to determine novel single-cell enzyme kinetics for three different cell types (HeLa, 293T, Jurkat). A kinetic model of this process predicted this varied response was due to variation in the concentration of carboxylesterase between cell types. Nordihydroguaiaretic acid (NDGA) was also characterized as an inhibitor of carboxylesterases. For HeLa cells, 20 nM of the 50 nM total carboxylesterases was unaffected by NDGA. This type of analysis could be directly applied to quantify a variety of intracellular enzymes with available fluorogenic substrates.
TL;DR: The association of aptamer-functionalized Pt- NPs to a throm bin aptamer/thrombin complex associated with an electrode allowed the amplified, electrocatalytic detection of thrombin with a sensitivity limit corresponding to 1 x 10(-9) M.
Abstract: Nucleic acid-functionalized Pt nanoparticles (Pt-NPs) act as catalytic labels for the amplified electrochemical detection of DNA hybridization and aptamer/protein recognition. Hybridization of the nucleic acid-modified Pt-NPs with a sensing nucleic acid/analyte DNA complex associated with an electrode enables the amperometric, amplified, detection of the DNA by the Pt NP electrocatalyzed reduction of H2O2 (sensitivity limit, 1 × 10-11 M). Similarly, the association of aptamer-functionalized Pt- NPs to a thrombin aptamer/thrombin complex associated with an electrode allowed the amplified, electrocatalytic detection of thrombin with a sensitivity limit corresponding to 1 × 10-9 M.
TL;DR: Scanning electron microscopy images indicate significant improvement in the microstructure of the proposed electrode compared to carbon paste electrodes, which allows sensitive, low-potential, simple,low-cost, and stable electrochemical sensing of biomolecules and other electroactive compounds.
Abstract: Ionic liquid, n-octylpyridinum hexafluorophosphate (OPFP) has been used to fabricate a new carbon composite electrode with very attractive electrochemical behavior. This type of carbon electrode has been constructed using graphite mixed with OPFP as the binder. The electrode has combined advantages of edge plane characteristics of carbon nanotubes and edge plane pyrolytic graphite electrodes together with the low cost of carbon paste electrodes and robustness of metallic electrodes. It provides a remarkable increase in the rate of electron transfer of different organic and inorganic electroactive compounds and offers a marked decrease in the overvoltage for biomolecules such as NADH, dopamine, and ascorbic acid. It also circumvents NADH surface fouling effects as well as furnishing higher current density for a wide range of compounds tested. Depending on the choice of the electrolyte, the electrode can have the ion-exchange property and adsorptive characteristics of clay-modified electrodes. The proposed electrode thus allows sensitive, low-potential, simple, low-cost, and stable electrochemical sensing of biomolecules and other electroactive compounds. Scanning electron microscopy images indicate significant improvement in the microstructure of the proposed electrode compared to carbon paste electrodes. Such abilities promote new opportunities for a wide range of electrochemical and biosensing applications.
TL;DR: The development of a low-cost, sensitive colorimetric sensor array for the detection and identification of volatile organic compounds (VOCs) is reported, and by proper choice of dyes and substrate, the array is essentially nonresponsive to changes in humidity.
Abstract: The development of a low-cost, sensitive colorimetric sensor array for the detection and identification of volatile organic compounds (VOCs) is reported. Using an array composed of chemoresponsive dyes, enormous discriminatory power is possible in a simple device that can be imaged easily with an ordinary flatbed scanner. Excellent differentiation of closely related organic compounds can be achieved, and a library of 100 VOCs is presented. The array discriminates among VOCs by probing a wide range of intermolecular interactions, including Lewis acid/base, Bronsted acid/base, metal ion coordination, hydrogen bonding, and dipolar interactions. Importantly, by proper choice of dyes and substrate, the array is essentially nonresponsive to changes in humidity.
TL;DR: An accessible method to form a planar lipid bilayer, simply by contacting two monolayers assembled at the interface between water and organic solvent in a microfluidic chip is described.
Abstract: Artificial planar lipid bilayers are a powerful tool for the functional study of membrane proteins, yet they have not been widely used due to their low stability and reproducibility. This paper describes an accessible method to form a planar lipid bilayer, simply by contacting two monolayers assembled at the interface between water and organic solvent in a microfluidic chip. The membrane of an organic solvent containing phospholipids at the interface was confirmed to be a bilayer by the capacitance measurement and by measuring the ion channel signal from reconstituted antibiotic peptides. We present two different designs for bilayer formation. One equips two circular wells connected, in which the water/solvent/water interface was formed by simply injecting a water droplet into each well. Another equips the cross-shaped microfluidic channel. In the latter design, formation of the interface at the sectional area was controlled by external syringe pumps. Both methods are extremely simple and reproducible, es...
TL;DR: The approach demonstrated the feasibility of using the developed nanosensor for rapid determination of Hg(II) in aqueous environmental samples and in batteries.
Abstract: A new gold-nanoparticle (AuNP)-based sensor for detecting Hg(II) ions in aqueous solution has been developed. Rhodamine B (RB) molecules that are highly fluorescent in bulk solution fluoresce weakly when they are adsorbed onto AuNP surfaces as a result of fluorescence resonance energy transfer and collision with AuNPs. In the presence of metal ions such as Hg(II), RB molecules are released from the AuNP surface and thus restore the florescence of RB. The modulation of the photoluminescence quenching efficiency of RB−AuNPs in the presence of Hg(II) ions can achieve a large turn-on fluorescence enhancement (400-fold) in aqueous solution, and the entire detection takes less than 10 min. We have improved the selectivity of the probe further by modifying the AuNP surfaces with thiol ligands (mercaptopropionic acid, mercaptosuccinic acid, and homocystine) and adding a chelating ligand (2,6-pyridinedicarboxylic acid) to the sample solutions. Under the optimum conditions, the selectivity of this system for Hg(II)...
TL;DR: The developed PDDA/AChE/PDDA/CNT/GC biosensor integrated into a flow injection system was used to monitor organophosphate pesticides and nerve agents, such as paraoxon and had excellent operational lifetime stability with no decrease in the activity of enzymes for more than 20 repeated measurements over a 1-week period.
Abstract: A highly sensitive flow injection amperometric biosensor for organophosphate pesticides and nerve agents based on self-assembled acetylcholinesterase (AChE) on a carbon nanotube (CNT)-modified glassy carbon (GC) electrode is described. AChE is immobilized on the negatively charged CNT surface by alternatively assembling a cationic poly(diallyldimethylammonium chloride) (PDDA) layer and an AChE layer. Transmission electron microscopy images confirm the formation of layer-by-layer nanostructures on carboxyl-functionalized CNTs. Fourier transform infrared reflectance spectrum indicates the AChE was immobilized successfully on the CNT/PDDA surface. The unique sandwich-like structure (PDDA/AChE/PDDA) on the CNT surface formed by self-assembling provides a favorable microenvironment to keep the bioactivity of AChE. The electrocatalytic activity of CNT leads to a greatly improved electrochemical detection of the enzymatically generated thiocholine product, including a low oxidation overvoltage (+150 mV), higher sensitivity, and stability. The developed PDDA/AChE/PDDA/CNT/GC biosensor integrated into a flow injection system was used to monitor organophosphate pesticides and nerve agents, such as paraoxon. The sensor performance, including inhibition time and regeneration conditions, was optimized with respect to operating conditions. Under the optimal conditions, the biosensor was used to measure as low as 0.4 pM paraoxon with a 6-min inhibition time. The biosensor had excellent operational lifetime stability with no decrease in the activity of enzymes for more than 20 repeated measurements over a 1-week period. The developed biosensor system is an ideal tool for online monitoring of organophosphate pesticides and nerve agents.
TL;DR: A novel two-nanoparticle assay with aptamers as the molecular recognition element for leukemia cells is developed using a cell-based SELEX strategy and allows for specific recognition of the cells from complex mixtures including whole blood samples.
Abstract: We have developed a method for the rapid collection and detection of leukemia cells using a novel two-nanoparticle assay with aptamers as the molecular recognition element. An aptamer sequence was selected using a cell-based SELEX strategy in our laboratory for CCRF-CEM acute leukemia cells that, when applied in this method, allows for specific recognition of the cells from complex mixtures including whole blood samples. Aptamer-modified magnetic nanoparticles were used for target cell extraction, while aptamer-modified fluorescent nanoparticles were simultaneously added for sensitive cell detection. Combining two types of nanoparticles allows for rapid, selective, and sensitive detection not possible by using either particle alone. Fluorescent nanoparticles amplify the signal intensity corresponding to a single aptamer binding event, resulting in improved sensitivity over methods using individual dye-labeled probes. In addition, aptamer-modified magnetic nanoparticles allow for rapid extraction of target...
TL;DR: This method for the determination of cocaine, amphetamines, morphine, cannabinoids, methadone, and some of their metabolites in wastewater could be useful to estimate and monitor drug consumption in the population in real time, helping social scientists and authorities to combat drug abuse.
Abstract: Residues of illicit drugs and their metabolites that are excreted by humans may flow into and through wastewater treatment plants. The aim of this study was to develop a method for the determination of cocaine, amphetamines, morphine, cannabinoids, methadone, and some of their metabolites in wastewater. Composite 24-h samples from urban treatment plants were enriched with deuterated internal standards before solid-phase extraction. High-pressure liquid chromatography tandem mass spectrometry with multiple reaction monitoring was used for quantitation. Recoveries were generally higher than 80%, and limits of quantifications were in the low nanograms-per-liter range for untreated and treated wastewater. The overall variability of the method was lower than 10% for untreated and 5% for treated wastewater. The method was applied to wastewater samples coming from two treatment plants in Italy and Switzerland. Quantification ranges were found to be 0.2−1 μg/L for cocaine and its metabolite benzoylecgonine, 80−20...
TL;DR: Microfluidics is utilized to isolate picogram and subpicogram mRNA templates, as well as to synthesize cDNA from these templates, which form the foundation for highly parallel single-cell gene expression studies.
Abstract: Single-cell gene expression analysis holds great promise for studying diverse biological systems, but methodology to process these precious samples in a reproducible, quantitative, and parallel fashion remains challenging. Here, we utilize microfluidics to isolate picogram and subpicogram mRNA templates, as well as to synthesize cDNA from these templates. We demonstrate single-cell mRNA isolation and cDNA synthesis, provide quantitative calibrations for each step in the process, and measure gene expression in individual cells. The techniques presented here form the foundation for highly parallel single-cell gene expression studies.
TL;DR: Novel data analysis software, XCMS, was used to monitor all metabolite features detected from an array of serum extraction methods, with application to metabolite profiling using electrospray liquid chromatography/mass spectrometry (ESI-LC/MS).
Abstract: The aim of metabolite profiling is to monitor all metabolites within a biological sample for applications in basic biochemical research as well as pharmacokinetic studies and biomarker discovery. Here, novel data analysis software, XCMS, was used to monitor all metabolite features detected from an array of serum extraction methods, with application to metabolite profiling using electrospray liquid chromatography/mass spectrometry (ESI-LC/MS). The XCMS software enabled the comparison of methods with regard to reproducibility, the number and type of metabolite features detected, and the similarity of these features between different extraction methods. Extraction efficiency with regard to metabolite feature hydrophobicity was examined through the generation of unique feature density distribution plots, displaying feature distribution along chromatographic time. Hierarchical clustering was performed to highlight similarities in the metabolite features observed between the extraction methods. Protein extracti...
TL;DR: The automated analysis of total lipid extracts was powered by a robotic nanoflow ion source and produced currently the most detailed description of the glycerophospholipidome.
Abstract: We report a method for the identification and quantification of glycerophospholipid molecular species that is based on the simultaneous automated acquisition and processing of 41 precursor ion spectra, specific for acyl anions of common fatty acids moieties and several lipid class-specific fragment ions. Absolute quantification of identified species was linear within a concentration range of 10 nM−100 μM and was achieved by spiking into total lipid extracts a set of synthetic lipid standards with diheptadecanoyl (17:0/17:0) fatty acid moieties, representing six common classes of glycerophospholipids. The automated analysis of total lipid extracts was powered by a robotic nanoflow ion source and produced currently the most detailed description of the glycerophospholipidome.
TL;DR: This paper presents the first in vivo application of surface-enhanced Raman scattering (SERS), which was used to obtain quantitative in vivo glucose measurements from an animal model.
Abstract: This paper presents the first in vivo application of surface-enhanced Raman scattering (SERS). SERS was used to obtain quantitative in vivo glucose measurements from an animal model. Silver film over nanosphere surfaces were functionalized with a two-component self-assembled monolayer, and subcutaneously implanted in a Sprague−Dawley rat such that the glucose concentration of the interstitial fluid could be measured by spectroscopically addressing the sensor through an optical window. The sensor had relatively low error (RMSEC = 7.46 mg/dL (0.41 mM) and RMSEP = 53.42 mg/dL (2.97 mM).
TL;DR: Novel high-throughput sample preparation strategies for MALDI imaging mass spectrometry (IMS) and profiling are presented and an acoustic reagent multispotter was developed to provide improved reproducibility for depositing matrix onto a sample surface, for example, a tissue section.
Abstract: Novel high-throughput sample preparation strategies for MALDI imaging mass spectrometry (IMS) and profiling are presented. An acoustic reagent multispotter was developed to provide improved reproducibility for depositing matrix onto a sample surface, for example, such as a tissue section. The unique design of the acoustic droplet ejector and its optimization for depositing matrix solution are discussed. Since it does not contain a capillary or nozzle for fluid ejection, issues with clogging of these orifices are avoided. Automated matrix deposition provides better control of conditions affecting protein extraction and matrix crystallization with the ability to deposit matrix accurately onto small surface features. For tissue sections, matrix spots of 180−200 μm in diameter were obtained and a procedure is described for generating coordinate files readable by a mass spectrometer to permit automated profile acquisition. Mass spectral quality and reproducibility was found to be better than that obtained with...
TL;DR: It is shown that diffusion occurs because of the concentration gradient between the compounds in solution flowing through the interstices and the stagnant liquid within the pores of the packing, and transport of these compounds into the pores results.
Abstract: Nowadays, there is a lot of buzz in separations science about the porous materials known as monoliths. Monoliths are used mostly as separation media and supports, and each one can be roughly compared to a single porous ''particle''. Monoliths are an alternative to columns packed with particulate stationary phases, which chromatographers have used for >100 years. Although that technology is well developed, it is not flawless. Packed columns have rather large void volumes; this is an inevitable result of the particulate character of the packing. Even in a perfectly organized array of monodisperse spheres, {approx}30% of the volume is interstitial voids. In reality, the percentage is even larger. As a consequence, a significant part of the column volume is not used for separation. However, this is not the only problem. Consider what happens when a mobile phase is pumped through a column packed with a standard porous packing. The liquid flows readily through the interstitial voids between the particles, where resistance to its flow is the smallest (Figure 1a). In contrast, the liquid in the pores remains stagnant. If a sample is injected into the stream of the mobile phase, the compounds in the sample will also be carried throughmore » the voids. However, diffusion occurs because of the concentration gradient between the compounds in solution flowing through the interstices and the stagnant liquid within the pores of the packing, and transport of these compounds into the pores results. Once the concentration ''pulse'' has passed by the bead, the compounds diffuse back from the pores into the surrounding liquid; eventually, only the original stagnant phase remains within the pores.« less