Introduction
Recycling Preparative HPLC is a separation technique that can achieve complete separation of a number of components by repeatedly cycling the eluent to the same column. This has the same effect as using longer or multiple columns. When applied to preparative HPLC each purified component can be fractionated very effectively.
This technique has the following features:
(1) Reduced column costs, as it is not necessary to use longer or multiple columns.
(2) Reduces environmental loads and costs, because in recycle mode, the same mobile phase solvent is used repeatedly while improving separation.
(3) Effective preparative purification because each component is fractionated only after improving the separation.
LC-4000 Semi-preparative HPLC system
Experimental
Chromatographic conditions
Column: Megapak GEL 201C (20 mm ID x 500 mmL)
Eluent: Chloroform
Eluent Flow Rate: 3.0 mL/min
Detection: UV detection (254 nm)
Injection Volume: 200 μL
Standard Sample: Polystyrene Oligomer (1.0 %(w/v) in Chroloform)
Figure 1. Flow Diagram of Recycling Preparative HPLC system
Results
Figure 2 shows recycle chromatogram of a Polystyrene Oligomer, showing the improvement of separation by recycling six times in approximately 180 minutes. Three components tetramer, pentamer and hexamer of Polystyrene Oligomer were clearly separated and fractionated.
Figure 2. Recycle Chromatogram of Polystyrene Oligomer
