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 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.
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 (10/90/0.1) B) acetonitrile/water/TFA (50/50/0.1) 25-45%B (0-40 min) |
Temperature | ambient |
Detection | UV at 214 nm |