Mass spectrometric identification of the N-monosubstituted N-hydroxylamino functionality in protonated analytes via ion/molecule reactions in tandem mass spectrometry.

RATIONALE

N-Monosubstituted hydroxylamines correspond to an important class of metabolites for many bioactive molecules. In this study, a tandem mass spectrometric method based on ion/molecule reactions was developed for the identification of compounds with the N-monosubstituted hydroxylamino functionality.

METHODS

The diagnostic ion/molecule reaction occurs between protonated analytes with 2-methoxypropene (MOP) inside a linear quadrupole ion trap mass spectrometer.

RESULTS

Most protonated compounds with N-monosubstituted and disubstituted hydroxylamino and oxime functional groups react with MOP via proton transfer and formation of a stable adduct in a linear quadrupole ion trap mass spectrometer. However, only protonated compounds with N-monosubstituted hydroxylamino groups form the characteristic MOP adduct-MeOH product. Possible mechanisms of this reaction are discussed.

CONCLUSIONS

A method based on functional group-selective ion/molecule reactions in a linear quadrupole ion trap mass spectrometer has been demonstrated to allow the identification of protonated compounds with the N-monosubstituted hydroxylamino functionality. Only N-monosubstituted hydroxylamines react with MOP via formation of an adduct that has eliminated methanol.