Feasibility of ultra-performance liquid chromatography-ion mobility-time-of-flight mass spectrometry in analyzing oxysterols

- We have developed a new method for the analysis of oxysterols by UPLC-IM-TOFMS.
- p-toluenesulfonyl isocyanate derivatization improved specificity and sensitivity.
- The method showed good quantitative performance.

Oxysterols are oxygenated cholesterols that are important in many cell functions and they may also be indicative of certain diseases. The purpose of this work was to study the feasibility of ultra-performance liquid chromatography-ion mobility-time-of-flight mass spectrometry (UPLC-IM-TOFMS) using traveling wave cell in analyzing oxysterols and especially their isomers in biological samples. Oxysterols were analyzed as their p-toluenesulfonyl isocyanate derivatives, which improved the separation of isomeric oxysterols by ion mobility and ionization efficiency in the electrospray ionization step. The UPLC-IM-TOFMS method was shown to be fast and to provide good quantitative performance. The feasibility of the method was demonstrated in the analyses of oxysterols in fibroblast cell samples.