How to troubleshoot split peaks on an ACQUITY UPLC column - WKB28044

OBJECTIVE or GOAL

Troubleshoot split peaks on an ACQUITY UPLC column.

ENVIRONMENT

• ACQUITY UPLC column
• Split peaks

PROCEDURE

1. Put a fresh column on the system, and add a guard column if appropriate. 
2. Inject benchmarking standard to confirm that the fresh column shows good peak shape. 
3. Run your normal mobile phase conditions through the column for as many cycles as it takes to see the split peaks; make no injections. 
4. After the mobile phase cycles are complete, inject the benchmarking standard again. Does the peak split? If yes, there is contamination in the flow path coming from the reservoirs through to the column. Take steps to decontaminate the system. If no, move on to the next step.
5. Inject blank solution that contains the liquid components of your sample diluent (e.g., water only, water and organic, or water and buffer).
6. Run the mobile phase conditions with the injected blanks for as many cycles as it takes to see the split peaks.
7. Does the peak split after this exercise?  If yes, the injector is contaminated. Take steps to clean the injector.

ADDITIONAL INFORMATION

For additional information, see following application notes:

• Improving Method Robustness for Routine Analysis of Pharmaceutical Formulations
• USP Method Transfer of Donepezil Tablets from HPLC to UPLC
• Controlling Contamination in LC/MS Systems - Best Practices

Also see the article Split peaks seen on an ACQUITY UPLC system.