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How to perform a solvent changeover procedure on Waters Styragel columns to convert from DMF to DMSO - WKB50557

Article number: 50557

OBJECTIVE or GOAL

Convert Styragel columns packed in dimethylformamide (DMF) to dimethylsulfoxide (DMSO).

ENVIRONMENT

  • Gel permeation chromatography (GPC)
  • Styragel
  • Solvent changeover

PROCEDURE

  1. Set the column oven to 60–80 °C.
  2. For 7.8 x 300 mm columns,  set the pump flow rate to 0.1 mL/min throughout this procedure.
    • For 6.0 x 150 mm columns, set the pump flow rate to 0.06 mL/min throughout this procedure.
    • For 4.6 x 300 mm columns, set the pump flow rate to 0.03 mL/min throughout this procedure.
  3. If you are running a gradient, program a 95:5 DMF/DMSO to 100% DMSO over four hours.
    • If you are running in Isocratic mode,  make separate DMF/DMSO solutions in 5% steps from 95:5  DMF/DMSO to 100% DMSO, and increase DMSO strength over four hours.
  4. Flush with 100% DMSO for six hours per column. If there are two columns in the column bank, flush for 12 hours. This equates to 10 column volumes per column.

This solvent switch should be performed only once in the column's lifetime.

ADDITIONAL INFORMATION

GPC Columns – Operating Procedures:

  • Always ramp from 0.1 mL/min to approximately 1 mL/min over five minutes.
  • Do likewise when ramping down flow rate.
  • Typical operating flow rates when performing GPC separations on Styragel columns:
    • 7.8 x 300 mm columns  -  1.0 mL/min
    • 6.0 x 150 mm columns  -  0.6 mL/min
    • 4.6 x 300 mm columns  -  0.3 mL/min
  • When the column is not in use but is kept on a system, keep the flow rate at 0.1 mL/min minimum at all times. Do not power-off the pump.

id50557, EMPGPC, GPCV2K

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