HPLC Analysis of Protein Standards on Zenix® SEC-300 by Size Exclusion As Affected by Arginine
Materials
analytical column
Zenix® SEC-300 Gel Filtration Column, 3 μm
L × I.D. 30 cm × 4.6 mm, 3 μm particle size, stainless steelaccessory
mobile phase component
L-Arginine
from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%CONDITIONS
column
Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 μm particles (A777028)
mobile phase
[A], 0.15 M potassium phosphate, monobasic, pH 7.0 with potassium hydroxide; [B] 0.15 M potassium phosphate, monobasic, 0.2 M arginine, pH 7.0 with phosphoric acid (mobile phase is either 100% A or 100% B)
flow rate
0.3 mL/min
pressure
1030 psi 100% A (71 bar)
pressure
1165 psi 100% B (80 bar)
column temp.
25 °C
detector
UV, 280 nm
injection
2.5 μL
sample
Waters BEH200 SEC protein standard
Description
Analysis Note
Arginine can have benefitial effects on peak shape and recovery of hydrophobic proteins when chromatographed by size exclusion chromatography. In this case, with protein standards that are not particularly hydrophobic, no dramatic affect is noted, as assessed by peak efficiency, peak widths, or peak retention.
Legal Information
Zenix is a registered trademark of Sepax Technologies