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G005795

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

application for HPLC

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About This Item

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)

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

Description
    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

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