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- Posts: 35
- Joined: Sun Apr 08, 2018 3:17 am
I'm developing and trying to validate a HILIC method using IPA as my mobile phase with 5% Water.
I developed this method on a Waters Acquity H Class system using 1.0mL/min and had no problems. I recently tried re-visiting this and now I'm having all kinds of problems.
Obviously I washed the system with 100% Water with a union to flush the system of any residual salts.
When I try to run the chromatographic parameters listed below I'm getting these cyclical peaks and valleys now.
I read somewhere that you have to use lower flow rates for IPA due to the increase in viscosity, but even when I try that I'm still getting cyclical baseline disturbances.
Any suggestions?
Column: Luna Amino 150mm X 4.6mm, 5um
Mobile phase: 95:5 IPA:Water
Diluent: Mobile phase
Injection volume: 50uL
Detection wavelength 210nm
Flow Rate: Tried 1.0mL/min then going to 0.5mL/min and now trying 0.25mL/min
Column Teperature: Ambient.