Coupling ultra high-pressure liquid chromatography with mass spectrometry: constraints and possible applications.
The introduction of columns packed with porous sub-2μm particles and the extension of the upper pressure limit of HPLC instrumentation to 1300bar (ultra-high pressure liquid chromatography, UHPLC) has opened new frontiers in resolution and speed of analysis. However, certain constraints appear when coupling UHPLC technology with mass spectrometry (MS). First, the most significant limitation is related to the narrow peaks that are produced by UHPLC that require a fast duty cycle, which is only available on the latest generations of MS devices. Thus, certain analyzers are more readily compatible with UHPLC (e.g., QqQ or TOF/MS) than others (e.g., ion trap or FT-MS). Second, due to the reduction of the column volume, extra-column band broadening can become significant, leading to a reduction in the kinetic performance of the UHPLC-MS configuration. Third, as the mobile phase linear velocity is higher in UHPLC, the electrospray ionization source must also be able to provide high sensitivity at flow rates of up to 1mL/min. Despite these limitations, the UHPLC-MS/MS platform has successfully been employed over the last decade for various types of applications, including those related to bioanalysis, drug metabolism, multi-residue screening, metabolomics, biopharmaceuticals and polar compounds.
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Boulevard d'Yvoy 20, 1211 Geneva 4, Switzerland.
Marta
M
Rodriguez-Aller
Robert
R
Gurny
Jean-Luc
JL
Veuthey
Davy
D
Guillarme
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D055432:Metabolomics / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism; D010944:Plants / Q000737:chemistry / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000295:instrumentation / Q000379:methods*
D004364:Pharmaceutical Preparations