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 (α) chmotrypsinogen A remains at approximately 3 until the sample load reaches 1.0 mg. Even when the sample load is 5.0 mg, the Rs is approximately 1.1, which means the substances can still be separated.


Sample :
Ribonuclease A
α-Chymotrypsinogen A

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