How do I transfer an ACQUITY UPLC method/column to an HPLC method/column? - WKB231157

OBJECTIVE or GOAL

Transfer an ACQUITY UPLC method that uses an ACQUITY UPLC column to an HPLC system that uses an HPLC column.

ENVIRONMENT

• Method transfer
• UPLC column
• HPLC column
• ACQUITY

PROCEDURE

1. Choose an HPLC column with the same bonded phase as the ACQUITY column, except in a larger particle size.
2. Measure the system dwell volume for each system.
3. Specify the original method information on the left-hand side of the Columns Calculator Version 2.0, including system dwell volume.
4. Specify the HPLC column dimensions, particle size, and the HPLC system dwell volume on the right-hand side of the calculator.
5. The calculator will calculate the scaled injection volume, flow rate, and gradient for the HPLC column.

ADDITIONAL INFORMATION

Maintaining a similar "L/Dp" value for the original column vs. the new column will help to maintain similar resolution on the transferred method.

If a method is transferred to a column that has a lower L/Dp value vs. the original column, you will have less resolution in the transferred method.

If a method is transferred to a column that has a higher L/Dp value vs. the original column, you will have more resolution in the transferred method.

(L/Dp = Length over Particle Diameter)

See:  L/dp Comparison Chart For LC Columns

Also see:

Waters Columns Calculator User Guide Infographic

Dwell Volume and Extra-Column Volume: What Are They and How Do They Impact Method Transfer

Successful Transfer of Size-Exclusion Separations between HPLC and UPLC

Transferring RapiFluor-MS Labeled N-Glycan HILIC Separations Between UPLC and HPLC

Fundamentals of a Successful LC Method Transfer - Advancing from HPLC to UPLC

Transferring Compendial HPLC Methods to UPLC Technology