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Merck
CN

G1006596

HPLC Analysis of Proteins on TSKgel® BioAssist S, Ion Selectivity Comparison

application for HPLC

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technique(s)

HPLC: suitable

test parameters

column: TSKgel BioAssist S, 5 cm x 4.6 mm I.D., 7 μm particles (819686)
mobile phase: [A] 10 mM potassium phosphate, dibasic, pH 7 with phosphoric acid; [B] 0.3 M potassium potassium phosphate, pH 7 with phosphoric acid; ([C] 10 mM sodium phosphate, dibasic, pH 7 with phosphoric acid; [D] 0.3 M sodium potassium phosphate, dibasic, pH 7 with phosphoric acid;)
gradient: 100% A to 100% B (or 100% C to 100% D) in 33 min, held at 100% B (or D) for 5 minutes
flow rate: 0.3 mL/min
pressure: 615 psi total pressure (42 bar total pressure)
column temp.: 25 °C
detector: UV, 280 nm
injection: 5 μL
sample: g/L ea: ribonuclease a, 0.38; cytochrome c, 0.15; lysozyme, 0.23; a-chymptrypsinogen, 0.23

suitability

application for HPLC

application(s)

life science and biopharma

Analysis Note

The choice of exchangable counterion can affect retention and selectivity when performing ion exchange chromatography of proteins. This application note compares sodium and potassium gradients for changes in retention and selectivity.

Legal Information

TSKgel is a registered trademark of Tosoh Corporation

Analyte

Description
    Ribonuclease A from bovine pancreas, Type I-A, powder, ≥60% RNase A basis (SDS-PAGE), ≥50 Kunitz units/mg protein
    α-Chymotrypsinogen A from bovine pancreas, essentially salt-free, lyophilized powder
    Cytochrome c from bovine heart, ≥95% based on Mol. Wt. 12,327 basis, powder, suitable for mammalian cell culture
    Lysozyme from chicken egg white, powder or granules, ≥39,000 units/mg protein

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