All SEC peaks are getting broader on ACQUITY H-Class Bio - WKB73253

SYMPTOMS

• All SEC peaks are getting broader over time or showing double peaks
• Resolution of separation is no longer good

ENVIRONMENT

• ACQUITY UPLC H-Class Bio
• ACQUITY UPLC
• Size-exclusion chromatography

CAUSE

Microbial growth on column

FIX or WORKAROUND

1. Remove the contaminated solvent bottles and column.
2. Clean your solvent lines, pump, and injector using phosphoric acid wash (30%). See Controlling Contamination in LC/MS Systems.
3. Clean your detector according to the instructions in its operator manual (e.g., How to clean the ACQUITY PDA Detector flow cell).
4. Rinse the entire system with UPLC-grade water in a clean glass borosilicate bottle.
5. Make fresh solvent in clean bottles, fit a new SEC column, and set up the system with your SEC application according to the care and use manual of the column (ACQUITY UPLC Protein BEH SEC Columns and Standards).
6. To avoid microbial growth, follow guidelines in Guidelines for routine use and maintenance of ultra-performance size-exclusion and ion-exchange chromatography systems and Size exclusion and ion-exchange chromatography of proteins using the ACQUITY UPLC H-Class System.

ADDITIONAL INFORMATION