UPLC-ToF MS analysis | | UPV

Título: UPLC-ToF MS analysis
Descripción: Separation of compounds by Ultra-high Performance Liquid Chromatography and detection by means of Time of Flight Mass Spectrometry. López Gresa, MP.; Caballero Vizcaino, M. (2019). UPLC-ToF MS analysis. hdl.handle.net/10251/117811
Descripción automática: In this video, the host explains the process and benefits of using ultra-performance liquid chromatography (UPLC) coupled with time of flight mass spectrometry (MS) for the analysis of polar and highly polar compounds, taking tomato leaves as an example. The combination of UPLC and MS, including high-pressure liquid chromatography (HPLC) techniques, is highlighted as particularly effective for complex substances that cannot be measured by gas chromatography (GC). The discussion covers the separation process in detail, touching on how compounds interact with the stationary phase, the importance of high-resolution, and the role of the mobile phase in transporting the sample through the column.
The presenter outlines the components of the HPLC instrument, including a degasser, sampler, column, detector, and data management system. Special focus is given to the two types of columns-normal and reversed-phase-with the latter being more commonly used with UPLC, as it works well with non-polar stationary phases (such as C18) and polar mobile phases (like methanol and water).
The video further details the detection process, comparing various types of detectors and their applications in quantitative analysis, including photodiode array (PDA), fluorescence, and mass spectrometry. The procedure for untargeted metabolomics analysis using UPLC-PDA-MS is also described, including the preparation and settings of the UPLC system.
Finally, mass spectrometry is introduced as a powerful technique for studying metabolites, with the video explaining the fundamental principles of ionization methods and mass analysis. The specific procedures for mass spectrometer calibration and the importance of maintaining stable temperatures during analysis are emphasized. The conclusion underscores UPLC coupled with time of flight mass spectrometry as optimal for analyzing polar and semi-polar compounds, with its effectiveness demonstrated by separating methanol extracts from tomato leaves.
Autor/a: López Gresa María Pilar
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#UPLC-MS #2302 - Bioquímica

[@shaniton1519]

Beautiful instrument