HPLC Analysis of Insulin (Species Variants) on Ascentis® Express Phenyl-Hexyl

HPLC Analysis of Insulin (Species Variants) on Ascentis® Express Phenyl-Hexyl

Conditions

column Ascentis Express Phenyl-Hexyl, 15 cm x 2.1 mm, 2.7 µm particles (53338-U)
column temp. 30 °C
mobile phase [A] 70:30, water with 0.1% TFA:acetonitrile with 0.09% TFA; [B] 55:45, water with 0.1% TFA:acetonitrile with 0.09% TFA
gradient 0 to 100% B in 7.5 min
flow rate 0.3 mL/min
pressure 3960 psi (273 bar)
sample 200 µg/mL in 0.1% formic acid
injection 0.5 µL
detector UV, 215 nm

Description

Analysis Note These three insulins have more than 76% identical positions in a primary structure alignment. The human and porcine proteins are most similar with 85% identity. The Phenyl-Hexyl phase provides an alternate retention and selectivity to a C18 phase, particularly the C18 phase on a larger pore silica, Ascentis Express ES-C18
Categories Analytical Chromatography, Proteins, Peptides, Hormones
Featured Industry Life Science and Biopharma
Pharmaceutical (small molecule)
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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