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)
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
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.
Legal Information
TSKgel is a registered trademark of Tosoh Corporation