Betaine aldehyde reacts selecWvely with hydroxyl group via nucleophilic addiWon to form a hemiacetal salt with a stable posiWve charge on quaternary ammonium ion. That is why it could be used for derivaWsaWon of reWnol, vitamin D2 and D3, L-ascorbic acid, glucose and cholesterol. This feature was shown for the first Wme by Wu et al. with the DESI ion source . Our study showed that cholesterol analysis with this derivaWsaWon is possible also with MALDI ion source . In this study, fast and simple protocol for glucose derivaWzaWon by betaine aldehyde was proposed. Moreover, our study showed some discrepancies between ion images obtained by NEDC, comparing to betaine aldehyde derivaWves, which requires consideraWon and the utmost a]enWon.