How do I change solvents on an Ultrahydrogel column? - WKB107044

OBJECTIVE or GOAL

Change solvents on an Ultrahydrogel column while minimizing damage to the packing material.

ENVIRONMENT

• High Performance Liquid Chromatography (HPLC)
• Gel permeation chromatography (GPC)
• Size exclusion chromatography (SEC)
• Gel filtration chromatography (GFC)
• Columns
• Water-soluble polymers
• Solvent switching
• Solvent changeover

PROCEDURE

1. Start flow rate at 0.05 mL/min and monitor the baseline. Continue flowing at 0.05 mL/min until your baseline stabilizes.
2. Increase flow rate to 0.10 mL/min and monitor the baseline. Continue flowing at 0.10 mL/min until your baseline stabilizes.
3. Increase flow rate to 0.15 mL/min and monitor the baseline. Continue flowing at 0.15 mL/min until your baseline stabilizes.
4. Increase flow rate to 0.20 mL/min and monitor the baseline. Continue flowing at 0.20 mL/min until your baseline stabilizes.
5. Increase flow rate to 0.25 mL/min and monitor the baseline. Continue flowing at 0.25 mL/min until your baseline stabilizes.
6. Increase flow rate to 0.30 mL/min and monitor the baseline. Continue flowing at 0.30 mL/min until your baseline stabilizes.
7. Equilibrate column bank equal to 36 mL per 7.8 x 300 mm column installed on system at 0.3mL/min.  
8. After equilibration, increase flow in increments of 0.05 mL/min, ensuring that you see baseline stability before each ramp-up.
9. Continue this process until you reach an operational flow rate of no more than 1.0 mL/min.

ADDITIONAL INFORMATION

Waters Gel Permeation Chromatography (GPC) Capabilities Brochure

Ultrahydrogel Columns Care & Use Manual