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Enhanced Characterization of Metabolites using Microflow UPLC-MS and Collision Cross Section Measurements

Waters
25 Jun 2014

Metabolites of Midazolam were produced from incubations in mouse, rat and human hepatocytes. The assay is based on the direct analysis of the extracts produced by protein precipitation and subsequent analysis using a new microfluidic device combining an ultra high pressure liquid chromatography (UPLC) analytical column with a nano-flow electrospray source. The ceramic device contains a 5 cm analytical column of 150 µm internal diameter along with the ionization emitter in one interchangeable unit. All microfluidic, gas and electrical connections are automatically engaged when the ceramic microfluidic device is inserted into the source enclosure. All experiments were performed on a Waters Synapt G2-Si Q-tof mass spectrometer operating in MSE data acquisition mode. Detected metabolites of interest were subject to separation by ion mobility spectrometry and the CCS calculated for all the isomeric forms detected.

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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.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. Ion Mobility SpectrometryASMSThe American Society for Mass Spectrometry (ASMS) is an organization dedicated to advancing the science and application of mass spectrometry. ASMS supports professionals through events, publications, and research initiatives, promoting innovation in areas like proteomics, metabolomics, and environmental analysis.

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Enhanced Characterization of Metabolites using Microflow UPLC-MS and Collision Cross Section Measurements