Perfluorinated acid alternatives to trifluoroacetic acid for reversed-phase high-performance liquid chromatography

J Chromatogr A. 1996 Oct 11;746(2):277-81. doi: 10.1016/0021-9673(96)00377-9.

Abstract

Over the past decade trifluoroacetic acid (TFA) has become the ion-pairing agent (IPA) of choice for reversed-phase high-performance liquid chromatography (RP-HPLC) of peptides and proteins. Reagent grade TFA is highly pure, water soluble, transparent at 220 nm and readily volatile. A drawback of this universal appeal is that several alternative perfluorinated carboxylic acids tend to be overlooked when TFA does not work in a particular separation. Examples are given comparing TFA selectivity with those of pentafluoropropionic acid, heptafluorobutyric acid, perfluoropentanoic acid, perfluorohexanoic acid and perfluoroheptanoic acid. We have found that increasing the IPA n-alkyl chain length can resolve sample components that otherwise co-elute in the void volume of TFA-based RP-HPLC. Examples are given for the resolution of an oligoglycine series and enhanced selectivity for a bovine hypothalamic extract.

MeSH terms

  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid / methods*
  • Fluoroacetates / chemistry*
  • Hypothalamus / chemistry
  • Indicators and Reagents
  • Oligopeptides / analysis
  • Reference Standards
  • Spectrophotometry, Ultraviolet
  • Trifluoroacetic Acid / chemistry*

Substances

  • Fluoroacetates
  • Indicators and Reagents
  • Oligopeptides
  • Trifluoroacetic Acid