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

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

Materials

Analytical column

Product No.
Description
Pricing

Ascentis® Express Phenyl-Hexyl, 2.7 μm HPLC Column

2.7 μm particle size, L × I.D. 15 cm × 2.1 mm

Standard

Product No.
Description
Pricing

Insulin from porcine pancreas

powder, ≥27 USP units/mg

Insulin from bovine pancreas

≥25 USP units/mg (HPLC), powder

CONDITIONS

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

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