How do I prepare mobile phases for HILIC applications? - WKB31055
OBJECTIVE or GOAL
Prepare mobile phase for HILIC applications.
ENVIRONMENT
- Hydrophilic Interaction Chromatography (HILIC)
- Mobile phase preparation
PROCEDURE
Low pH
Prepare a 200 mM stock buffer of ammonium formate by dissolving 12.6 g of formic acid ammonium salt and adding it to 900 mL of water in a 1 L volumetric flask. Twenty five milliliters of formic acid was added. Water was then added to the 1 L mark. Final solution pH was 3.0.
If using a Binary Solvent Manager system, prepare the low pH A and B mobile phases as follows:
- Mobile phase A: 90/10 ACN/water with 10 mM ammonium formate and 0.125% formic acid: Add 50 mL of 200 mM stock buffer and 50 mL of MilliQ water to 900 mL of ACN. Sonicate in warm water bath to eliminate cloudiness.
- Mobile phase B: 50/50 ACN/water with 10 mM ammonium formate and 0.125% formic acid: Add 50 mL of 200 mM stock buffer and 450 mL of water to 500 mL of ACN.
High pH
Prepare a 200 mM stock buffer of ammonium acetate by dissolving 15.4 g of ammonium acetate and adding it to 900 mL of water in a 1 L volumetric flask. Eight milliliters of ammonium hydroxide was added. Water was then added to the 1 L mark. The final solution pH was 9.0.
If using a Binary Solvent Manager system, prepare the high pH A and B mobile phases as follows:
- Mobile phase A: 90/10 ACN/water with 10 mM ammonium acetate and 0.04% ammonium hydroxide: Add 50 mL of 200 mM stock buffer and 50 mL of MilliQ water to 900 mL of ACN.
- Mobile phase B: 50/50 ACN/water with 10 mM ammonium acetate and 0.04% ammonium hydroxide: Add 50 mL of 200 mM stock buffer and 450 mL of water to 500 mL of ACN.
ADDITIONAL INFORMATION
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