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G005794

HPLC Analysis of Insulin (Species Variants) on BIOshell A160 Peptide C18

application for HPLC

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About This Item

technique(s)

HPLC: suitable

test parameters

column: BIOshell A160 Peptide C18, 15 cm x 2.1 mm I.D., 2.7 μm particles (66905-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: 4040 psi (278 bar)
column temp.: 30 °C
detector: UV, 215 nm
injection: 0.5 μL
sample: 200 μg/mL insulin (bovine, human, porcine) 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. Baseline resolution of these insulins in achieved on BIOshell A160 Peptide C18, with relative band spacing of these three peaks approximating their relative sequence similarities. BIOshell A160 Peptide C18 is ideally suited for RP chromatography of small proteins or large peptides.

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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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