ResourceSeparations

Bile Acid Profiling Using UltraPerformance Convergence Chromatography (UPC<sup>2</sup>) Coupled to ESI-MS/MS

3 Nov 2014

This application note demonstrates that the use of UltraPerformance Convergence (UPC2) in conjunction with ESI-MS/MS enables the simultaneous profiling of 25 bile acids including conjugated forms in biological fluids. This application demonstrates that this novel separation method is highly suitable for determining the effects of bile acid levels in triglyceride, cholesterol, and glucose metabolism and has a potential to speed up the development of novel drug targets for metabolic diseases.

ACQUITY UPC² System

Waters

Convergence Chromatography with the ACQUITY UPC² System Bring streamlined workflow, expanded selectivity, and an orthogonal technique to your laboratory. Convergence chromatography (CC) is a broad-based, complementary analytical platform that is taking its place alongside of LC and GC as one of the three essential separation technologies for modern laboratory analysis. This technique provides another, reliable choice as an analytical platform that is orthogonal to reversed-phase LC (RPLC), while significantly streamlining the entire analytical workflow. In convergence chromatography, the separation is achieved by manipulating the density and composition of a supercritical fluid-based mobile phase. Because of the very high diffusivity of the mobile phase, can be achieved. Additionally, the diversity of stationary phase and mobile phase (co-solvent) options provides access to the largest selectivity space available to any separation technique. The ACQUITY UPC² System is a holistically designed UltraPerformance Convergence Chromatography (UPC²) system that utilizes liquid CO2 as the primary mobile phase. The system leverages the chromatographic principles and selectivity of normal-phase chromatography while providing the ease-of-use of RPLC, enabling you to address routine and complex separations challenges. ACQUITY UPC² System: Discover what's been missing from your analytical toolbox With the ACQUITY UPC² System, scientists can now separate, analyze, and understand compounds that have challenged all other LC and GC technologies including: Hydrophobic and chiral compounds Lipids Thermally-labile samples Polymers It's also the perfect complement to MS due to its low solvent load and high resolution, narrow peaks and fast separations. ACQUITY UPC² System Features: Compressed carbon dioxide (CO2), the primary mobile phase, is less costly and less toxic than liquid mobile phases or carrier gases. Variable volume injection delivers 0.5 µL to 10 µL as well as the ability to match column and injection volume with minimal sample loss and need to change loops. Higher throughput and productivity allows you to analyze more samples per day with faster re-equilibration of columns and shorter cycle times, increasing laboratory productivity. Co-solvent and column switching provides flexibility in method development for faster solvent screening and faster column screening.

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Xevo TQ-S

Waters

Xevo TQ-S tandem quadrupole mass spectrometer - designed for the most demanding quantitative UPLC/MS/MS applications. The ultimate in tandem quadrupole performance allows you to achieve unrivalled sensitivity and robustness, available to everyone due to the accessibility of the Xevo platform. The Xevo TQ-S gives you class leading sensitivity and robustness, qualitative information when you need it most and accelerated method development for sensitive and robust methods. Reducing complexity, increasing ease of use, and ensuring the correct result every time.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. Supercritical Fluid ChromatographySupercritical Fluid Chromatography (SFC) is used for the rapid separation and analysis of complex mixtures. SFC systems require injectors, pumps, columns, ovens and detectors. The SFC retention method uses a supercritical fluid, usually CO2. There are a number of different SFC detector types including UV / VIs, ECD, mass spectrometry and fluorescence.UltraPerformance Convergence ChromatographyConvergence Chromatography expands the capabilities of reversed phase LC and conventional GC techniques. By varying mobile phase strength, pressure, temperature and stationary phase Convergence Chromatography facilitates the separation, detection and quantification of many compounds and samples. The CO<sub>2< / sub> mobile phase used reduces the need for toxic solvents.MetabolitesMetabolites are the intermediate and end products of metabolism. Analyzing metabolites offers insight into cellular processes and disease mechanisms and is critical in fields like pharmacology, toxicology, and diagnostics. Explore our peer-reviewed product directory to find the best tools for metabolite analysis, compare products, check reviews, and get pricing directly from manufacturers.MetabolomicsMetabolomics is the study of small metabolites (the intermediates and products of metabolism). It involves the identification and quantification of cellular metabolites using analytical technologies such as GC, HPLC, NMR, and LC/MS.Drug MetabolismDrug metabolism is the enzymatic conversion within the body of pharmaceutical substances into other chemical compounds. The primary site for drug metabolism is the liver.