Ion Mobility Shift Reagents for Lipid Double Bonds Based on Paterno-Buchi Photoderivatization with Halogenated Acetophenones

The Paterno-Buchi (PB) reaction is a cycloaddition reaction between a carbon-carbon double bond (C=C) and a photochemically excited carbonyl-containing compound. The constrained ring formed between the C=C bond and the PB reagent is more susceptible to fragmentation by collision-induced dissociation, which facilitates identification of the C=C position within the fatty acyl tails of lipids. Although the original PB reaction using acetone had a low yield of derivatized lipids and therefore a low yield of diagnostic ions, a new generation of PB reagents based on halogenated acetophenones has improved the reaction yield substantially. In this study, we investigated the use of halogenated PB reagents and ion mobility to improve the identification of PB-derivatized lipids by shifting them out of the densely populated lipid region of ion mobility-mass spectrometry (IM-MS) space. Several halogenated PB reagents containing fluorine, chlorine and bromine were investigated for their ability to decrease the collision cross-section (CCS) values of derivatized lipids and yield sufficient intensity for both the derivatized lipid and its diagnostic ions. We found that 4'-chloro-2',6'-difluoroacetophenone (CDFAP) displayed the best performance, with an average decrease in CCS of 4.4% and yield of derivatized lipids and diagnostic ions comparable to the trifluorinated acetophenone reagent proposed by the Xia group. The unique isotope pattern resulting from the chlorine substituent aided in identification of the derivatized lipids and their diagnostic ions, as well. We further demonstrate that derivatization with CDFAP preserves the separation of lipids classes in IM-MS space.

Automated summary

The use of halogenated PB reagents and ion mobility technique improves the identification of PB-derivatized lipids. 4'-chloro-2', 6'-difluoroacetophenone (CDFAP) shows the best performance, reducing the collision cross-section (CCS) values of derivatized lipids and yielding sufficient intensity for both the derivatized lipid and its diagnostic ions.