HPLC Analysis of Proteins by Cation Exchange on Proteomix® WCX: Affect of Particle Size

HPLC Analysis of Proteins by Cation Exchange on Proteomix® WCX: Affect of Particle Size

Conditions

column Proteomix WCX-NP1.7, 5 cm x 4.6 mm I.D., 5 µm particles (Z777177)
column Proteomix WCX-NP3, 5 cm x 4.6 mm I.D., 5 µm particles (Z777185)
column Proteomix WCX-NP5, 5 cm x 4.6 mm I.D., 5 µm particles (Z777193)
column temp. 25 °C
mobile phase [A] 10 mM potassium phosphate, dibasic, pH 7 with phosphoric acid; [B] 0.3 M potassium phosphate, dibasic, pH 7 with phosphoric acid
gradient 0 to 100% B in 10 min, held at 100% B for 5 minutes
flow rate 0.5 mL/min
pressure 700 psi, 1200 psi, 3400 psi total pressure, respectively (48 bar, 83 bar, 234 bar total pressure, respectively)
sample g/L ea: ribonuclease a, 0.38; cytochrome c, 0.15; lysozyme, 0.23; a-chymptrypsinogen, 0.23; in mobile phase A
injection 5 µL
detector UV, 280 nm

Description

Analysis Note Smaller particles can affect retention and peak widths in ion exchange chromatogaphy of proteins. Smaller particles provide higher total surface area, lower peak widths, and higher backpressure. With higher resolution afforded by the smaller particle size, greater chromatographic detail is evident with minor contaminants.
Categories Analytical Chromatography, Proteins
Featured Industry Life Science and Biopharma
Legal Information Proteomix is a registered trademark of Sepax Technologies
suitability application for HPLC

Materials

     
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