REGENERATING A REVERSED PHASE COLUMN

Requirements:

Time: 4 Hrs
Reagents: Water, DMSO, MeOH, CHCl₃
Flow rate: 0.7 ml/min
Procedure:

1. Fix the column in the normal direction.
   
2. Run the water for 10 minutes at 55ºC.
   
3. Inject 4 injections of DMSO (100µL each) at 2 minutes time interval.
   
4. Wash the column with water for half an hour.
   
5. Run the MeOH for one hour through the column in reverse direction.
   
6. Run the CHCl3 for one hour through the column in reverse direction.
   
7. Again Run the MeOH for one hour in reverse direction.
   
   
8. Finally run the MeOH for 20 minutes in normal direction.
   
9. Check the column performance.