HILIC UHPLC-MS/MS for fast and sensitive bioanalysis: accounting for matrix effects in method development
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Updated: May 7, 2026
Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
Published on: May 20, 2013
Lucie Havlíková1, Hana Vlčková, Petr Solich
1Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic.
Background:
Matrix effects are considered to be a main obstacle of quantitative bioanalytical LC-MS/MS methods. Therefore it is often required to minimize them in order to increase method reliability. HILIC has been referenced as one of possible approaches. However, there is a lack of experimental evidence in scientific literature so far.
Methodology:
Matrix effects were evaluated using spiked serum samples after SPE and protein precipitation prior to UHPLC-ESI-MS/MS. Chromatography was performed in both HILIC and reversed-phase mode. The influence of the matrix effects on the signal response was assessed using a set of 34 compounds of pharmaceutical interest and post-extraction addition approach.
Results:
The advantages and drawbacks of the HILIC and reversed-phase chromatographic modes were compared and discussed in detail.
Conclusion:
HILIC demonstrated the potential to reduce the occurrence of matrix effects when a more thorough sample pretreatment procedure such as SPE was applied.
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