HPLC Analysis of Insulin Variants and Analogs on BIOshell™ A160 Peptide C18

HPLC Analysis of Insulin Variants and Analogs on BIOshell™ A160 Peptide C18

Conditions

column BIOshell A160 Peptide C18, 15 cm x 2.1 mm, 2.7 µm particles (66905-U)
column temp. 75 °C
mobile phase [A] 75:25 water (0.1% trifluoroacetic acid) : acetonitrile (0.1% trifluoroacetic acid); [B] 50:50 water (0.1% trifluoroacetic acid) : acetonitrile (0.1% trifluoroacetic acid); (67:33, A:B)
flow rate 0.5 mL/min
pressure 3600 psi (248 bar)
sample mix of six insulin variants, 100 µg/mL in 0.1% trifluoroacetic acid
injection 0.5 µL
detector UV, 215 nm

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.
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 HPLC

Materials

     
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