What are the common causes of having little or no retention and/or peak shape problems on a HILIC column? - WKB235914
ENVIRONMENT
- Hydrophilic interaction chromatography (HILIC)
- Bridged ethylene hybrid (BEH)
- ACQUITY Premier
- ACQUITY UPLC Amide
- Atlantis Premier
- BEH Z-HILIC
- CORTECS
ANSWER
Retention problems may result from:
- Improper column conditioning out of the box, prior to first injection.
- Using a reverse-phase gradient (low organic to high organic).
- Improper column re-equilibration at the end of each gradient.
Peak shape problems may result from:
- Improper column conditioning out of the box, prior to first injection.
- Strong sample solvent effects.
- Poor connection in the flow path (can occur for any type of LC column).
ADDITIONAL INFORMATION
Failing to fully condition a HILIC column before first injection may result in poor retention and/or peak shape problems.
Forgetting to assign the correct mobile phase bottles for HILIC separations can also occur by accident. In HILIC, you always begin a separation with ~95% acetonitrile (weak solvent) and ~5% aqueous buffered solution (strong solvent). Then you elute with increasing aqueous. Alternative polar solvents may also be used for elution.
Atlantis Premier BEH Z-HILIC Columns Care and Use Manual
ACQUITY UPLC and ACQUITY PREMIER BEH Columns Care and Use Manual
XBridge XP and XBridge PREMIER 2.5 μm BEH Columns Care and Use Manual
XBridge Columns Care & Use Manual
CORTECS 1.6um Columns Care & Use Manual
CORTECS 2.7um Columns Care & Use Manual
Webinars:
Approaches to Polar Retention | 3-Part Webinar Series:
Part 1: Tips & Tricks to Retain and Separate Polar Compounds Using Reverse Phase Liquid Chromatography
Video: Benefits of Atlantis Premier BEH Z-HILIC Columns
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