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

UHPLC/MS Analysis of Insulin Variants and Analogs on BIOshell™ A160 Peptide C18 Using Ammonium Formate 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 (8 mM ammonium formate, pH 2.6 with formic acid:acetonitrile); [B] 50:50 (8 mM ammonium formate, pH 2.6 with formic acid:acetonitrile); (77:23, A:B)
flow rate 0.2 mL/min
sample Mixture of six insulin variants, 100 µg/mL, 8 mM ammonium formate, pH 2.6 with formic acid
injection 0.5 µL
detector MS, ESI(+), 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 ammonium formate as a mobile phase modifier yields more efficient peaks and less signal suppression during electrospray ionization (ESI) than some traditional ion-pair reagents (trifluoroacetic acid, formic acid).
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

     
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