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

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

Conditions

column TSKgel BioAssist S, 5 cm x 4.6 mm I.D., 7 µm particles (819686)
column temp. 25 C
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 psitotal pressure (42 bar total pressure)
sample g/L ea: ribonuclease a, 0.38; cytochrome c, 0.15; lysozyme, 0.23; a-chymptrypsinogen, 0.23
injection 5 µl
detector UV, 280 nm

Description

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.
Categories Analytical Chromatography, Proteins
Featured Industry Life Science and Biopharma
Legal Information TSKgel is a registered trademark of Tosoh Corporation
suitability application for HPLC

Materials

     
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