How do I make a focused gradient? - WKB27535

OBJECTIVE or GOAL

To design a focused gradient in order to further resolve impurities.

ENVIRONMENT

• Prep
• Preparative Chromatography
• Autopurification
• Columns
• OBD
• Gradient
• isolation

PROCEDURE

1. Determine the system dwell volume on prep system.
2. Run scouting gradient on analytical column of similar or same chemistry as used in prep system.
3. Design the focused gradient based on the analytical run.
4. Run the new focused gradient on the scaled prep column.

ADDITIONAL INFORMATION

For suggested tests and calculations to figure out dwell volume and focused gradients, see the following:

Developing Focused Gradients for Isolation and Purification