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Why is trifluoroacetic acid (TFA) used in reverse-phase chromatography for protein purification?

Posted June 1, 2020


Trifluoroacetic acid (TFA) is used as an ion paring agent in reversed-phase chromatography to improve peak shape and resolution. The silica used in columns may have metal ion impurities, which can cause peak tailing and loss of resolution. An ion-pair reagent such as TFA added to the mobile phase at a concentration of ~0.1% can “shield” the metal ion and help to maintain good peak shape.

Additional resources

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Miller, J. M. (2003). Chromatography. digital Encyclopedia of Applied Physics, 1055-1102.

Janson, J. C. (Ed.). (2012). Protein purification: principles, high resolution methods, and applications (Vol. 151). John Wiley & Sons.