How do I clean an ACQUITY UPLC BEH SEC column or an XBridge BEH SEC column? - WKB70819

ENVIRONMENT

• Size exclusion chromatography (SEC)
• BEH SEC columns

ANSWER

Cleaning options include the following:

1. Flush column with 10 column volumes of a concentrated salt solution at low pH (for example, 0.5M Na₂SO₄, pH 2.7)
2. Flush column with 10 column volumes of a low concentration (e.g. 10%) of methanol in UPLC-grade water (for ACQUITY) or HPLC-grade water (for XBridge)..

NOTE: Avoid use of ionic detergents and other surfactants if the SEC column will be used to analyze native proteins.

ADDITIONAL INFORMATION

Also see:

How should I clean an ACQUITY BEH SEC column or XBridge BEH SEC column if I suspect protein has precipitated on the column?

Can I clean XBridge BEH SEC columns and ACQUITY UPLC BEH SEC Columns with injections of DMSO (dimethylsulfoxide)?

Can I clean an XBridge BEH SEC Column or ACQUITY UPLC BEH SEC column with Guanidine or Urea?

ACQUITY UPLC BEH SEC Columns  and Standards Care & Use Manual

XBridge Protein BEH SEC Columns & Standards Care & Use Manual

Note: Choose a cleaning solvent on the basis of sample properties (e.g., use option "a" to remove basic protein and option "b" to remove hydrophobic proteins).
Chaotropic agents can solvate strongly adsorbed proteins via hydrogen bond disruption.

As a last resort, you can try flow reversal or back flushing at a low flow rate (e.g., 0.1 mL/min). However, this approach may further damage the column or only provide short-lived improvement in performance.