HPLC Analysis of Protein Standards on Zenix® SEC-300 by Size Exclusion As Affected by Sodium Perchlorate
Materials
Analytical column
Zenix® SEC-300 Gel Filtration Column, 3 μm
L × I.D. 30 cm × 4.6 mm, 3 μm particle size, stainless steelStandard
CONDITIONS
column
Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 μm particles (Z777028)
mobile phase
[A], 0.1 M sodium phosphate, monobasic, pH 7.0 with sodium hydroxide; [B] 0.1 M sodium phosphate, monobasic, 0.1 M sodium perchlorate, pH 7.0 with sodium hydroxide; (mobile phase is either 100% A or 100% B)
flow rate
0.3 mL/min
pressure
1040 psi (72 bar)
column temp.
25 °C
detector
UV, 215 nm
injection
2.5 μL
sample
Waters BEH200 SEC protein standard
Description
Analysis Note
Perchlorate is an alternative to chloride, to inspect possible benefits of a higher ionic strength mobile phase for size exclusion chromatography. Higher ionic strength mobile phases may enhance chromatographic performance by reducing ion-exchange activity of the column. Perchlorate is compatible with conventional U/HPLC instrumentation with stainless steel hardware/tubing, and generally shows greater elution strength than chloride, as a displacer anion. With this protein standard, addition of perchlorate to the mobile phase does result in some improvements to peak shape and selectivity.
Legal Information
Zenix is a registered trademark of Sepax Technologies