HPLC Analysis of Proteins by Cation Exchange on Proteomix® WCX: Affect of Particle Size
Materials
related product
Proteomix® WCX-NP1.7 Weak Cation Exchange Column
L × I.D. 5 cm × 2.1 mm, 1.7 μm particle size, stainless steelProteomix® WCX-NP3 Weak Cation Exchange Column
L × I.D. 5 cm × 4.6 mm, 3 μm particle size, stainless steelProteomix® WCX-NP5 Weak Cation Exchange Column
L × I.D. 5 cm × 4.6 mm, 5 μm particle size, stainless steelused together
CONDITIONS
column
Proteomix WCX-NP5, 5 cm x 4.6 mm I.D., 5 μm particles (Z777193)
column
Proteomix WCX-NP3, 5 cm x 4.6 mm I.D., 5 μm particles (Z777185)
column
Proteomix WCX-NP1.7, 5 cm x 4.6 mm I.D., 5 μm particles (Z777177)
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)
column temp.
25 °C
detector
UV, 280 nm
injection
5 μL
sample
g/L ea: ribonuclease a, 0.38; cytochrome c, 0.15; lysozyme, 0.23; a-chymptrypsinogen, 0.23; in mobile phase A
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.
Legal Information
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