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HPLC Analysis of Protein Standards on Zenix® SEC-300 by Size Exclusion As Affected by Sodium Perchlorate

HPLC Analysis of Protein Standards on Zenix® SEC-300 by Size Exclusion As Affected by Sodium Perchlorate

Conditions

column Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 µm particles (Z777028)
column temp. 25 C
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)
sample Waters BEH200 SEC protein standard
injection 2.5 µl
detector UV, 215 nm

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.
Categories Analytical Chromatography, Proteins
Featured Industry Life Science and Biopharma
Legal Information Zenix is a registered trademark of Sepax Technologies
suitability application for HPLC

Materials

     
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