How should I clean an ACQUITY BEH SEC column or XBridge BEH SEC column if I suspect protein has precipitated on the column? - WKB108019

ENVIRONMENT

• Ultra Performance Liquid Chromatography (UPLC)
• Ultra High Performance Liquid Chromatography (UHPLC)
• High Performance Liquid Chromatography (HPLC)
• Bridged ethylene hybrid (BEH)
• Size exclusion chromatography (SEC)
• Column
• XBridge BEH SEC column
• ACQUITY BEH SEC column
• Protein separation
• Column cleaning

ANSWER

1. Perform a shallow gradient from 0% to 100% acetonitrile over a period of 30 minutes, and perform this two times.
2. Inject a protein molecular weight standard to verify that step 1 corrects the issue.
3. If the issue is not corrected, choose one of the following options:
   1. Make three 100-µL injections 50:50 of DMSO:water (dimethylsulfoxide) if your system is compatible with DMSO.
   2. Make three 100-µL injections of 6M guanidnium chloride in a mobile phase of 50:50 H₂O/acetonitrile.

ADDITIONAL INFORMATION

Prior to cleaning the column, inject a protein molecular weight standard to confirm that the column is fouled.

Also see:

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