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Guide to HILIC (U)HPLC Columns

What Are HILIC Columns?

Hydrophilic Interaction Liquid Chromatography (HILIC) Columns are specialized (U)HPLC columns designed to retain and separate highly polar analytes. In contrast to reversed-phase methods, HILIC uses hydrophilic stationary phases and predominantly organic mobile phases to enhance retention of polar compounds—making it ideal for metabolites, polar drugs, sugars, and small peptides.

How Do HILIC Columns Work?

HILIC Columns operate via partitioning between a water-rich layer on the stationary phase and an organic-rich mobile phase. Polar analytes partition into the aqueous layer and elute later than less polar compounds. This mechanism improves retention and resolution for highly polar substances and reduces ion suppression in LC–ESI–MS workflows.

 

Applications of HILIC in HPLC & UHPLC

  • Polar metabolite analysis: Ideal for small molecules, peptides, nucleotides, and carbohydrates.

  • LC–MS workflows: High organic content mobile phases enhance MS-sensitivity and reduce ion suppression.

  • Complementary selectivity: HILIC retains polar analytes weakly retained by reversed-phase methods.

  • Bioanalysis & drug metabolism: Crucial for profiling polar metabolites in complex matrices.

Choosing the Right HILIC Column

  • Stationary phase type: Silica-based vs. zwitterionic vs. amide columns offer different selectivities.

  • Particle size and format: Choose analytical or UHPLC-compatible formats based on your system and resolution needs.

  • Mobile phase composition: High organic (e.g., 90 % acetonitrile) content improves retention and MS sensitivity.

  • pH and buffer considerations: Volatile buffers (formate, ammonium acetate) ensure compatibility with MS.

Why Use HILIC Columns?

HILIC Columns enable reliable retention of polar compounds, improved resolution for early-eluting analytes, and enhanced sensitivity in LC–ESI–MS, making them essential tools for modern analytical labs.

 

Need help selecting a HILIC (U)HPLC column? Contact YMC Europe for expert advice.

Contact us for consultation
  • YMC Europe GmbH
  • Phone +49 2064 427-0
  • Schöttmannshof 19
  • Fax +49 2064 427-222
  • D-46539 Dinslaken