1. Enhancing sensitivity of liquid chromatography–mass spectrometry of peptides and proteins using supercharging agents
- Author
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Nshanian, Michael, Lakshmanan, Rajeswari, Chen, Hao, Loo, Rachel R Ogorzalek, and Loo, Joseph A
- Subjects
Analytical Chemistry ,Chemical Sciences ,Physical Chemistry ,Liquid chromatography ,Electrospray ionization mechanism ,Supercharging ,Ion suppression ,electrospray ionization mechanism ,ion suppression ,liquid chromatography ,supercharging ,Organic Chemistry ,Physical Chemistry (incl. Structural) ,Analytical chemistry ,Physical chemistry ,Atomic ,molecular and optical physics - Abstract
Trifluoroacetic acid (TFA) is often used as a mobile phase modifier to enhance reversed phase chromatographic performance. TFA adjusts solution pH and is an ion-pairing agent, but it is not typically suitable for electrospray ionization-mass spectrometry (ESI-MS) and liquid chromatography/MS (LC/MS) because of its significant signal suppression. Supercharging agents elevate peptide and protein charge states in ESI, increasing tandem MS (MS/MS) efficiency. Here, LC/MS protein supercharging was effected by adding agents to LC mobile phase solvents. Significantly, the ionization suppression generally observed with TFA was, for the most part, rescued by supercharging agents, with improved separation efficiency (higher number of theoretical plates) and lowered detection limits.
- Published
- 2018