HPLC Analysis of Protein Standards on Zenix® SEC-300 by Size Exclusion As Affected by Potassium Chloride
Materials
analytical column
Zenix® SEC-300 Gel Filtration Column, 3 μm
L × I.D. 30 cm × 4.6 mm, 3 μm particle size, stainless steelaccessory
mobile phase component
standard
CONDITIONS
column
Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 μm particles (Z777028)
mobile phase
[A], 0.15 M potassium phosphate, monobasic, pH 7.0 with potassium hydroxide; [B] 0.15 M potassium phosphate, monobasic, 0.2 M potassium chloride, pH 7.0 with potassium hydroxide; (mobile phase is either 100% A or 100% B)
flow rate
0.3 mL/min
pressure
1000 psi (69 bar)
column temp.
25 °C
detector
UV, 215 nm
injection
2.5 μL
sample
Waters BEH200 SEC protein standard
Description
Analysis Note
In size exclusion chromatography, addition of halide salts (typically sodium or potassium chloride) in modest concentration, to an aqueous buffer, can yield improved chromatographic performance with protein analytes. In this case, as regards chromatographic performanc, little impact is noted except in the case of IgG, where peak shape is improved.
Legal Information
Zenix is a registered trademark of Sepax Technologies