Trifluoroacetic acid (TFA) is commonly used as mobile phase additive to improve retention and peak shape characteristics in hydrophilic interaction liquid chromatography (HILIC) of intact proteins. However, when using electrospray ionization-mass spectrometry (ESI-MS) detection, TFA causes ionization suppression and adduct formation leading to reduced analyte sensitivity. To overcome the latter issues, in this work we describe a membrane-based microfluidic chip with with multiple parallel channels (Microfluidic Esi Mobile Phase Ion Stripper, uEMPIS) for the selective post-column removal of TFA anions from HILIC eluents up to flow rates of 0.2 mL/min. An anion-exchange membrane was used to physically separate the column effluent from a stripper flow solution (running countercurrent) comprising acetonitrile, formic acid, and propionic acid. The exchange of ions via diffusion based on concentration gradient allowed the ion-exchange removal of TFA ions used during HILIC analysis of model proteins using mobile phases containing 0.1% TFA (13 mM). Separation selectivity and efficiency were maintained (with minor band broadening effects) while increasing the signal intensity and areas by improving ionization and reducing TFA adduct formation.
[doi:10.25345/C5V793]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: TFA removal ; Intact protein analysis ; ESI-MS
Principal Investigators: (in alphabetical order) |
Gargano Andrea, University of Amsterdam, Netherlands |
Submitting User: | agargan1 |
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