UHPLC/MS Analysis of Insulin Variants and Analogs on BIOshell™ A160 Peptide C18 Using Difluoroacetic Acid as a Mobile Phase Modifier

UHPLC/MS Analysis of Insulin Variants and Analogs on BIOshell™ A160 Peptide C18 Using Difluoroacetic Acid as a Mobile Phase Modifier

Conditions

column BIOshell A160 Peptide C18, 15 cm x 2.1 mm I.D., 2.7 µm particles (66905-U)
column temp. 75 °C
mobile phase [A] 75:25 water (0.1% v/v difluoroacetic acid): acetonitrile (0.1% v/v difluoroacetic acid); [B] 50:50 water (0.1% v/v difluoroacetic acid): acetonitrile (0.1% v/v difluoroacetic acid); (77:23, A:B)
flow rate 0.2 mL/min
sample Mixture of six insulin variants, 100 µg/mL, water (0.1% v/v difluoroacetic acid)
injection 0.5 µL
injection MS, ESI(+) TOF, TIC 100-3000 m/z

Description

Analysis Note BIOshell A160 Peptide C18 is an ideal column for reversed-phase separations of small proteins to large peptide sequences. There is over 75% sequence similarity between all six of the insulin variants in the sample; therefore, separation of these variants is achieved through the subtle differences in the polarities of surface residues on the peptides. The use of difluoroacetic acid (DFA) has less of an effect on ion suppression than trifluoroacetic acid (TFA), thus allowing for mass spectral detection of the insulin variants.
Categories Analytical Chromatography, Proteins, Peptides
Featured Industry Life Science and Biopharma
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
suitability application for UHPLC

Materials

     
Related Links