Heptafluorobutyric Acid (HFBA), HPLC Grade

HPLC Grade heptafluorobutyric acid (HFBA) is an ion-paired reagent specially developed for the separation of peptides and proteins by reverse-phase high-performance liquid chromatography. The reagent is in demand in proteomics, peptide analysis, preparative chromatography and in the identification of posttranslational modifications of proteins. HFBA belongs to the class of perfluorinated acids and is the most hydrophobic representative of this group. Compared with trifluoroacetic acid (TFA) and pentafluoropropionic acid (PFPA), HFBA provides a significantly stronger ion-pair interaction with positively charged amino acid residues (lysine, arginine, histidine). The resulting neutral complex increases the affinity of peptides to the hydrophobic stationary phase, which leads to an increase in retention time and an improvement in the shape of chromatographic peaks. The key advantage of HFBA is the ability to separate peptides that are not retained or eluted together in standard phases with TFA. Studies demonstrate that HFBA is excellent at separating glycated and non-glycated peptides, achieving basic resolution for some sequences where TFA provides only partial separation. In addition, HFBA effectively separates peptide mixtures by the number of charged groups, which makes it possible to separate peptides with a charge of +1 from peptides with a charge of +3 or +5. The product is characterized by high purity — the typical purity is 99.7 percent according to gas chromatography, the water content does not exceed 0.1 percent. The reagent is tested for sulfate content and UV absorption is measured at wavelengths of 230, 254 and 280 nanometers. Packaging is carried out in amber glass ampoules under nitrogen, which prevents oxidation and contamination. One ampoule with a volume of 1 milliliter allows you to prepare 1 liter of 0.1% HFBA solution for use as a mobile phase. The reagent is also used as a reagent for the sequencing of peptides and proteins by the Edman method, in solid-phase peptide synthesis and for the solubilization of peptides and proteins.