Technical Information

Overview of method development for isolation/purification Analytical scale conditions Loading Scale-up Preparative separation conditions Effective preparative separation technique Examples of preparative purification


Examples of preparative purification

Preparative purification was performed to obtain more than 200 mg of degradation product in total amount with over 95% purity for structural determination of the pharmaceutical degradation compound.

Analytical scale study

Separation method was developed on the analytical scale. Mobile phase pH was optimized in order to completely separate the target compound from the rest of the compounds. By applying the conditions for high resolution, loadability of a single run can be increased.

Eluent Buffer/acetonitrile (85/15)
Flow rate 0.8 mL/min
Temperature 35℃
Detection UV at 270 nm

Evaluation of temperature dependence

Separation conditions were studied at room temperature because the heated condition is not preferable on the scaled up purification. Even at room temperature, the target compound and the rest of the compounds were well separated and kept good peak shapes.

Eluent 20 mM CH3COOH (pH 3.2) /acetonitrile
Flow rate 0.8 mL/min
Detection UV at 270 nm
Injection 20 μL

Scale up and preparative separation

Based on the conditions studied on the analytical scale, column size and particle size were chosen. After the lab-scale preparation study, the large-scale preparation was performed.

Column YMC*GEL ODS-AQ (15 µm, 120 Å)
Eluent 20 mM CH3COOH/acetonitrile (85/15)
Temperature ambient
Detection UV at 270 nm