HILIC UHPLC-MS/MS for fast and sensitive bioanalysis: accounting for matrix effects in method development

Area of Science:

• Analytical Chemistry
• Bioanalysis
• Chromatography

Background:

• Matrix effects are a significant challenge in quantitative bioanalytical liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods.
• Minimizing matrix effects is crucial for improving the reliability of these analytical techniques.
• Hydrophilic Interaction Liquid Chromatography (HILIC) has been proposed as a potential strategy to mitigate matrix effects, but experimental validation is limited.

Purpose of the Study:

• To experimentally evaluate the effectiveness of HILIC compared to reversed-phase chromatography in reducing matrix effects.
• To assess the impact of HILIC on method reliability in bioanalytical LC-MS/MS.
• To investigate the combined effect of sample pretreatment (SPE) and HILIC on matrix effects.

Main Methods:

• Evaluation of matrix effects in spiked serum samples using ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS).
• Comparison of chromatographic separation using both HILIC and reversed-phase modes.
• Assessment of matrix effects on signal response for 34 pharmaceutical compounds using a post-extraction addition approach after solid-phase extraction (SPE) and protein precipitation.

Main Results:

• A detailed comparison of the advantages and disadvantages of HILIC and reversed-phase chromatographic modes was conducted.
• The study provided experimental evidence on the performance of HILIC in the context of matrix effects.

Conclusions:

• HILIC demonstrates potential for reducing matrix effects in quantitative bioanalytical LC-MS/MS.
• The efficacy of HILIC in mitigating matrix effects is enhanced when coupled with a robust sample pretreatment, such as solid-phase extraction (SPE).
• This study contributes experimental data supporting HILIC as a viable approach for improving bioanalytical method reliability by minimizing matrix effects.