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


Effective preparative separation techniques

Stacking injection

Separation time and the solvent consumption can be reduced by injecting the samples with overwrapping of the elution of the impurities (but not target compounds). Productivity is improved dramatically by using stacking injection.

Recycling chromatography

Recycling chromatography is a method that achieves the same effect as a separation using a longer column by repeated separation of the compounds with recirculation of the mobile phase. Purification can be performed under higher loading with recycling chromatography than with a single column method. In addition, consumption of solvents can be significantly reduced due to recirculation of mobile phase during the recycling process.

Effective second preparative separation

Re-purification may increase purity and/or recovery of the target compound when purity target or recovery target is not achieved by single purification cycle due to insufficient resolution.

Example of purification of peptide

Column YMC*GEL C4 (15 µm, 300 Å)
Eluent A) acetonitrile/water/TFA
B) acetonitrile/water/TFA
25-45%B (0-40 min)
Temperature ambient
Detection UV at 214 nm