Effect of Sample Load on Resolution (QA-825)

A mixture of transferrin and ovalbumin was injected into IEC QA-825 (a column for ion exchange chromatography) to obtain the relation between sample load and resolution(Rs). With up to 2mg of sample load, the Rs remained at approximately 3 with no decline. Rs was 2.5 when the sample load was 5mg. Even when the sample load was increased to 10 mg, Rs was approximately 2 with satisfactory resolution.

Sample :
Transferrin and Ovalbumin

Column       : Shodex IEC QA-825 (8.0mmID*75mm)
Eluent       : (A); 20mM Piperazine-HCl buffer(pH6.0)
               (B); (A) + 0.5M NaCl
               Linear gradient: 0min to 20min, (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.