Background: Hydrophilic interaction chromatography (HILIC) is becoming an increasingly popular alternative to traditional reversed-phase chromatography for the analysis of polar compounds. The ability to retain the most polar compounds in HILIC makes it attractive for the analysis of certain large groups of compounds, such as monoamines, which are inherently very polar.
Results: This paper details the development of a HILIC LC-MS/MS method for the analysis of monoamine neurotransmitters. The emphasis is on method development; in particular, the factors influencing sensitivity, peak shape and resolution. Mobile-phase ionic strength, temperature and stationary phase functionality are shown to be key parameters for the successful development of HILIC methods.
Conclusion: HILIC is shown to be an appropriate and suitable method for the analysis of monoamine neurotransmitters and an attractive alternative to reversed-phase analysis. The most polar analytes, which are essentially unretained by reversed-phase chromatography, demonstrate superior retention and resolution when analyzed by HILIC.