HPLC Analysis of Protein Standards by Size Exclusion on Zenix® as Affected by Pore Size: 100Å vs. 300Å

HPLC Analysis of Protein Standards by Size Exclusion on Zenix® as Affected by Pore Size: 100Å vs. 300Å

Conditions

column Zenix SEC-100, 30 cm x 4.6 mm I.D., 3 µm particles (Z777006)
column Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 µm particles (Z777028)
column Zenix SEC-100, 15 cm x 4.6 mm I.D., 3 µm particles
column Zenix SEC-300, 15 cm x 4.6 mm I.D., 3 µm particles (Z777026)
column temp. 25 °C
mobile phase 0.15 M sodium phosphate, monobasic, pH 7.0 with sodium hydroxide
flow rate 0.3 mL/min
pressure 1215 psi (84 bar)
sample Waters BEH125 SEC protein standard
injection 0.5 µL
detector UV, 215 nm

Description

Analysis Note With size exclusion chromatography, pore size fundamentally determines the relative retention of analytes. Larger pores permit inclusion of larger analytes, smaller pores exlcude larger analytes. This application compares the retention and selectivity of protein standards on columns of widely differing pore sizes as well as the sequential combination of both. The combination of the two in series provides an intermediate selectivity. Each 30 cm column was compared to combination of the two 15 cm columns connected in series; in the case of the latter, the SEC-300 preceding the SEC-100 column in the flow path.
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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