Approach to hydrophilic interaction chromatography column selection: application to neurotransmitters analysis

Area of Science:

• Analytical Chemistry
• Chromatography
• Separation Science

Background:

• Hydrophilic interaction liquid chromatography (HILIC) is a key technique for separating polar compounds.
• Understanding column performance is crucial for optimizing HILIC separations of complex mixtures like neurotransmitters.

Purpose of the Study:

• To compare the performance of 12 commercially available HILIC columns.
• To investigate factors influencing retention and selectivity for 12 neurotransmitters.
• To develop a generic procedure for optimizing HILIC analyses based on analyte ionization.

Main Methods:

• Comparative analysis of 12 HILIC columns.
• Retention and selectivity studies for 12 neurotransmitters across different compound classes (anionic, cationic, zwitterionic).
• Statistical analysis including factor analysis and principal component analysis (PCA) of retention data.

Main Results:

• Significant differences in separation behavior were observed among the HILIC stationary phases.
• PCA effectively distinguished column performance based on retention data for different compound types.
• Analyte ionization state was identified as a critical factor for HILIC method optimization.

Conclusions:

• The study provides valuable insights into HILIC column performance for neurotransmitter analysis.
• A statistical approach aids in understanding and predicting column selectivity.
• A generalized HILIC optimization strategy is proposed, considering analyte ionization states.