G005795

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

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: column: Ascentis Express Phenyl-Hexyl, 15 cm x 2.1 mm, 2.7 μm particles (53338-U); 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); column temp.: 30 °C; detector: UV, 215 nm; injection: 0.5 μL; sample: 200 μg/mL in 0.1% formic acid
• suitability: application for HPLC
• application(s): life science and biopharma; pharmaceutical (small molecule)

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

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

Analyte

Insulin from bovine pancreas, ≥25 USP units/mg (HPLC), powder
Insulin human, powder, recombinant, expressed in E. coli, suitable for cell culture
Insulin from porcine pancreas, powder, ≥27 USP units/mg