Effect of Sample Load on Resolution (CM-825)

When IEC CM-825 (a column forweak cation exchange chromatography) is used, the effect of sample load on the resolution is shown. The resolution (Rs) of a mixture of ribonuclease A and (alpha) chmotrypsinogen A remains at approximately 3 until the sample load reaches 1.0mg. Even when the sample load is 5.0mg, the Rs is approximately 1.1, which means the substances can still be separated.

Sample :
Ribonuclease A
alpha Chymotrypsinogen A

Column       : Shodex IEC CM-825 (8.0mmID*75mm)
Eluent       : (A); 20mM Sodium phosphate buffer(pH7.0)
               (B); (A) + 0.5M NaCl
               Linear gradient: 0min to 20min, (A) to (B)
Detector     : UV(280nm)
Column temp. : Room temp.