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