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