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Comprehensive Characterization of Monoclonal Antibodies Using a Microfluidic Chip-Q-TOF PlatformAccurate, sensitive, and comprehensive characterization of monoclonal antibodies is an absolute necessity to the pharmaceutical and diagnostic industries to meet regulatory requirements and ensure the efficacy and safety of the final product. Microfluidic chip-based high performance liquid chromatography technology interfaced with the mass accuracy of quadrupole time-of-flight mass spectrometry provides the ability to rapidly and efficiently assess the quality of intact monoclonal antibodies, confirm their amino acid sequence, and determine their glycosylation state, while consuming very small amounts of these precious products.
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Comparison of Extracts from Dry and Alcohol-Steamed Root of Polygonatum kingianum (Huang Jing) by Sub-2-µm–LC–TOF-MSMost plants used in traditional Chinese medicine must be processed before their medicinal usage; hence the effective ingredients may differ from those in the freshly harvested plant extracts. In this work, we present a fast and generic approach using sub-2-?m liquid chromatography–time-of-flight–mass spectrometry (sub-2-?m-LC–TOF-MS) coupled with multivariate statistical data analysis to systematically profile ingredient changes between fresh and processed samples of huang jing.
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High-Throughput Trace Analysis Using SERS-Coated Microtiter Plates with a Raman Plate ReaderSurface-enhanced Raman spectroscopy (SERS) has been studied extensively over the last few decades with many advances in preparation of SERS substrates and coatings. While the bulk of the research in SERS substrate preparation has been devoted to pushing detection limits to higher sensitivity for measurement of single samples, the application of SERS to high-throughput analysis has been largely ignored. In this article, we present the use of commercially available SERS-coated microtiter plates in a dedicated Raman microtiter plate reader, enabling high-throughput trace analysis measurements. This article also describes the SERS substrate, the high-throughput plate reader, and preliminary results from samples representing trace analysis of explosives, nerve agents, pharmaceuticals, and biological compounds.
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High-Throughput Trace Analysis Using SERS-Coated Microtiter Plates with a Raman Plate ReaderSurface-enhanced Raman spectroscopy (SERS) has been studied extensively over the last few decades with many advances in preparation of SERS substrates and coatings. While the bulk of the research in SERS substrate preparation has been devoted to pushing detection limits to higher sensitivity for measurement of single samples, the application of SERS to high-throughput analysis has been largely ignored. In this article, we present the use of commercially available SERS-coated microtiter plates in a dedicated Raman microtiter plate reader, enabling high-throughput trace analysis measurements. This article also describes the SERS substrate, the high-throughput plate reader, and preliminary results from samples representing trace analysis of explosives, nerve agents, pharmaceuticals, and biological compounds.
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A Mass Spectrometry GlossaryHere we present an updated glossary of industry terms - a handy guide for the seasoned analyst as well as those new to the field.
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Dried Blood Spots and High-Resolution Mass Spectrometry for Discovery Fast PK BioanalysisThis article introduces the advantages of accurate mass high-resolution mass spectrometry LC–MS (HRMS) coupled to the dried blood spot (DBS) technique for fast PK applications in a discovery environment. Compared with the established norm of plasma bioanalysis using triple quadrupoles, HRMS coupled to DBS is a viable alternative. The benefit is access to critical new information (HRMS bioanalysis) and significantly less stress on the animal (DBS), both factors that potentially improve the quality of early PK data.
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Confocal Raman AFM Imaging of PaperA better understanding of the micro- and nanostructures of cellulose fiber composition before and after various chemical treatments, together with the distribution of bleaching additives to the final paper composition, is essential for further improvements of paper properties. Confocal Raman imaging is a valuable tool for such studies, as it reveals not only optical information but also information regarding the 3D distribution of the chemical compounds in the submicrometer range. In the paper surface finishing process, additional surface roughness parameters can be measured by combining confocal Raman microscopy with atomic force microscopy. Such combined analytical microscopes allow a direct linking between high-resolution imaging and chemical identification of various species on a surface.
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Using LC–oa-TOF MSE with a Multivariate Statistical Sample Statistical Sample Profiling Strategy to Distinguish Chinese Red Ginseng from Korean Red GinsengBoth Chinese ginseng and Korean ginseng are similar plant species and undergo similar handling procedures when harvested and processed for sale. Despite their similarities, Korean ginseng commands a higher price than Chinese ginseng on the open market and is believed to produce different clinical effects than Chinese ginseng. Chinese researchers are now employing new techniques on the two varieties of ginseng to understand their chemical differences. HPLC/UV-based strategies for distinguishing the two types of ginseng have proven to be mostly ineffective due to lack of resolution. Using UltraPerformance liquid chromatography/orthogonal acceleration (oa)–TOF mass spectrometry and exact mass measurement, the authors developed a high-resolution method using multivariate statistical analysis for separating and identifying differences between Chinese ginseng and Korean ginseng at the molecular level.
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Vibrational Spectroscopic Discrimination of Herbal Medicines: Polygala senega, Polygala tenuifolia, and Glinus oppositifoliusThe noninvasive discrimination of powdered root material belonging to the Polygala genus and an adulterant is presented. The quality of the approach is assessed for attenuated total reflectance mid-infrared spectroscopy and diffuse reflectance near-infrared spectroscopy. Due to the pharmaceutical importance of Polygala related plant material, conclusions are drawn towards a laboratory independent discrimination of the samples.
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Advances in TOF-MS-Based Screening for Food Safety Residue Analysis with a Positive ApproachMass spectrometry plays an increasingly significant role in the analysis of residues and contaminants in food. Here we will illustrate how the combination of ultrahigh-pressure liquid chromatography (UHPLC) and high-resolution time-of-flight-mass spectrometry (TOF-MS) is used to generate a screen of veterinary drug residues in products of animal origin. The use of UHPLC–TOF-MS and dedicated, workflow directed software allows rapid screening for large numbers of residues and automated quantification of positive samples. In addition, we illustrate how the data generated using MSE acquisition mode enable critical structural information to be collected, which offers additional selectivity and confirmatory data for compound identification and facilitates elucidation of the structure of newly discovered compounds.
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Branching Out: Mass Spectrometry and the Shape of BiotherapeuticsThose fond of puns point out that mass spectrometry (MS) has become ever more focused in the last two decades, while at the same time offering ever more information. The dynamic market for biotherapeutics has driven a number of developments, particularly following the paradigm of well-characterized biopharmaceutical products (WCBP) (1,2). Partly as a result of automation and interfacing, those trained in biological or biochemical disciplines now use mass spectrometers routinely. This also means that the sorts of questions asked of MS have changed. Coping with biomolecule heterogeneity is a key challenge, not generally an issue for small molecule drugs. The data complexity means that mass information alone is insufficient. And at the submission stage, regulators are increasingly concerned about tertiary structure and conformation, something that was not previously an analytical requirement (2). Adding polyethylene glycol (PEG) to already heterogeneous molecules to prolong their half-lives in the body raises..
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Polarized Raman Spectroscopy of Aligned Semiconducting Single-Walled Carbon NanotubesAligned semiconducting single-walled carbon nanotubes (s-SWCNTs) are expected to outperform silicon as the next generation of integrated circuits. Greater utilization of polarized Raman spectroscopy is proving beneficial for efficient characterization of alignment in CNT films. Here, we present the results of how polarized Raman imaging can be used to effectively characterize alignment in large regions of aligned s-SWCNT films.
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Advances in TOF-MS-Based Screening for Food Safety Residue Analysis with a Positive ApproachMass spectrometry plays an increasingly significant role in the analysis of residues and contaminants in food. Here we will illustrate how the combination of ultrahigh-pressure liquid chromatography (UHPLC) and high-resolution time-of-flight-mass spectrometry (TOF-MS) is used to generate a screen of veterinary drug residues in products of animal origin. The use of UHPLC–TOF-MS and dedicated, workflow directed software allows rapid screening for large numbers of residues and automated quantification of positive samples. In addition, we illustrate how the data generated using MSE acquisition mode enable critical structural information to be collected, which offers additional selectivity and confirmatory data for compound identification and facilitates elucidation of the structure of newly discovered compounds.
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Improved Principal Component Discrimination of Commercial Inks Using Surface-Enhanced Resonant Raman ScatteringIn the three decades since its discovery, surface-enhanced Raman scattering (SERS) has been used in numerous applications to increase signal intensity in Raman scattering experiments. The current study provides insight into the more practical aspects of enhanced Raman sampling for laboratory users. We describe how the signal enhancement from a surface-enhanced resonant Raman scattering (SERRS) process improves the ability to discriminate between ink samples using principal component clustering.
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The Orbis Micro-XRF Analyzer SeriesBuilding on more than 10 years of Micro-XRF experience, the Orbis spectrometer yields a system with excellent Micro-XRF capability while setting a new standard in analytical flexibility. The Orbis incorporates a unique motorized turret integrating video and X-ray optics allowing coaxial sample view and X-ray analysis. The turret can accommodate two additional collimators along with the X-ray optic for a total of three X-ray beam sizes to expand the Orbis analytical capabilities beyond traditional Micro-XRF analysis. Primary beam filters can be used with all spot sizes available on the turret to allow true XRF analytical capabilities in a micro-spot analysis. The working distance is increased to allow analysis over rougher sample topography without sacrificing signal intensity.
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ADME/Pharmacokinetic Studies from Serum and Plasma: Improvements in Sample Preparation and LC–MS Analysis of Therapeutic OligonucleotidesThe authors discuss improvements in sample preparation for ADME/pharmacokinetic studies of therapeutic oligonucleotides.
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Increasing Productivity of ADME Studies Using Accurate Mass TechnologyA new time-of-flight mass spectrometer was evaluated for performing simultaneous metabolic stability measurement and metabolite identification with ultrahigh-pressure liquid chromatography. Six representative compounds (clomipramine, diclofenac, imipramine, haloperidol, verapamil, and midazolam) were incubated in rat liver microsomes at a more physiologically relevant substrate concentration (1 ?M). High-resolution full-scan and product-ion spectra were acquired in a single injection using generic methodology. Quantitative clearance of the parent was measured using the full-scan data. Major metabolites were identified using the accurate mass product ion spectra. High scanning speed allowed for a sufficient number of data points to be collected across the chromatographic peak for quantitative analysis. Sensitivity was sufficient for obtaining meaningful kinetics with a 1 ?M initial substrate concentration.
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Raman Spectroscopy of DocumentsDocuments have been investigated to determine the feasibility of utilizing Raman and SERS Raman spectroscopy for the identification and characterization of inks on paper. Fluorescence reduction methods have been employed to facilitate the analysis by reducing the nascent fluorescence from paper and ink. Furthermore, ink crossings were investigated to demonstrate that ink applied after creation of a document could be differentiated from the originally applied ink.
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ADME/Pharmacokinetic Studies from Serum and Plasma: Improvements in Sample Preparation and LC–MS Analysis of Therapeutic OligonucleotidesThe authors discuss improvements in sample preparation for ADME/pharmacokinetic studies of therapeutic oligonucleotides.
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High-Definition Screening for Boar Taint in Fatback Samples Using GC–MSHigh-definition screening by gas chromatography–mass spectrometry (GC–MS) is shown to be a viable option for the reliable identification of odorous compounds in pork.
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Metabolite Profiling Applications in Early Drug DiscoveryMetabolite profiling in drug discovery can contribute significantly at the lead optimization stage in two main application areas. The first is the identification of major metabolites, which provides medicinal chemists with information on the metabolic "soft spots." These soft spots are locations on the molecule particularly susceptible to metabolic modification, which can contribute to high pharmacokinetic clearance. This information then can be used to optimize the structure of a lead compound or chemical series to slow the rate of metabolism and therefore reduce hepatic clearance. This improves the absorption, distribution, metabolism, and excretion (ADME) properties of the compound, such as bioavailability, exposure (as measured by area under the curve), and half-life. Through iterative optimization of the structure and timely generation of metabolism data following each structural modification, pharmacokinetic properties can be improved while maintaining activity against the therapeutic target.
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The Chemical Analysis ProcessThe authors present an overview of the chemical analysis process.
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High-Throughput Trace Analysis Using SERS-Coated Microtiter Plates with a Raman Plate ReaderSurface-enhanced Raman spectroscopy (SERS) has been studied extensively over the last few decades with many advances in preparation of SERS substrates and coatings. While the bulk of the research in SERS substrate preparation has been devoted to pushing detection limits to higher sensitivity for measurement of single samples, the application of SERS to high-throughput analysis has been largely ignored. In this article, we present the use of commercially available SERS-coated microtiter plates in a dedicated Raman microtiter plate reader, enabling high-throughput trace analysis measurements. This article also describes the SERS substrate, the high-throughput plate reader, and preliminary results from samples representing trace analysis of explosives, nerve agents, pharmaceuticals, and biological compounds.
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Radio Frequency Glow Discharge Optical Emission Spectroscopy: Depth Profiling Analysis of Thin Anodic Alumina Films as Potential Reference Materials (PDF)The need for reference materials that can be applied in the area of thin films analysis has long been realized but is still, in general, under-addressed. Alumina films of single-micrometer thickness, having either fine distributions of impurities or delta function impurity marker layers, can be prepared routinely by anodic oxidation of electropolished aluminum specimens in appropriate electrolytes. Selected films were examined by transmission electron microscopy (TEM) and analyzed by radio frequency glow discharge optical emission spectroscopy (rf-GD-OES), providing very rapid, yet high-resolution, depth-resolved analysis of these electrically insulating materials.
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Carbon Nanotube Characterization by Resonance Raman SpectroscopyPhoton etc. has designed two narrowband tunable filters for resonance Raman spectroscopy.
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Moving Mid-IR Spectroscopy Forward in MedicineNick Stone of the University of Exeter explains why mid-infrared (mid-IR) spectroscopy is so valuable for spectral imaging in disease research and clinical diagnostics, and discusses his own recent work in this area.
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Fluorescence Spectroscopy: Theory and System DesignThis application note explores the fundamentals of fluorescence and different types of measurement setups.
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Improving the Analytical Workflow for Protein Biopharmaceutical Characterization with a Novel LC–MS System SolutionThis article presents an efficient analytical workflow for protein characterization using LC–MS.
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Low-Resolution Raman Spectroscopy in Science EducationRecent developments in photonics are finally making Raman instrumentation accessible to larger basic laboratories.
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Online Identification of Flavones from Flos Chrysanthemi by LC–MS-IT-TOFThe use of medicinal herbs as alternative treatment methods continues to grow. With this escalating use has come an increasing interest in determining the chemical compositions of these herbs in order to obtain a better understanding of their makeup and effects. In this study, Flos Chrysanthemi, a commonly used traditional Chinese medicine that has been cultivated for centuries, was analyzed to identify the main flavone compositions in one original breed of Flos Chrysanthemi (Hangbaiju) in China.
