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


METHODS
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.


CONCLUSIONS
HILIC demonstrated the potential to reduce the occurrence of matrix effects when a more thorough sample pretreatment procedure such as SPE was applied.
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic.
Lucie
L
Havlíková
Hana
H
Vlčková
Petr
P
Solich
Lucie
L
Nováková
D001798:Blood Proteins / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D057927:Hydrophobic and Hydrophilic Interactions; D004364:Pharmaceutical Preparations / Q000097:blood*; D052616:Solid Phase Extraction; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D001798:Blood Proteins; D004364:Pharmaceutical Preparations; D012997:Solvents