Convert HPLC Columns Between Reversed Phase and Normal Phase - How to
April 11, 2012
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Date: 11-APRIL-2012 Last Updated: 23-NOVEMBER-2025
Switching Between HPLC Modes: A Quick Guide
When working in different HPLC modes, a simple column-switching procedure can save time and ensure proper equilibration. The following 30-minute protocol allows you to transition between Reversed Phase (RP) and Normal Phase (NP) efficiently:
A – From Reversed Phase to Normal Phase
- Pump 100% methanol for 15 minutes at 1 mL/min.
- Follow with 100% methylene chloride for 15 minutes at the same flow rate.
- The column is now ready to be equilibrated with your NP-HPLC mobile phase.
B – From Normal Phase to Reversed Phase
- Pump 100% methylene chloride for 15 minutes at 1 mL/min.
- Follow with 100% methanol for 15 minutes at the same flow rate.
- The column is now ready to be equilibrated with your RP-HPLC mobile phase.
