Localization of double-bond positions in lipids by tandem mass spectrometry succeeding high-performance liquid chromatography with post-column derivatization

摘要: Rationale Lipids are a group of biomolecules known for their importance in broad variety biological tasks, which is attributed to the diversity molecular structures. The exact position double bond lipid affects its properties. Therefore, determination highly important. Nevertheless, unambiguous assignment still challenging task and requires development new approaches. Methods A novel method pinpointing double-bond positions lipids presented based on photochemical Paternὸ-Buchi reaction. reaction was performed using post-column derivatization by ultraviolet (UV) light irradiation (254 nm) an additional constant flow acetone. acetone adduct formed due with resulting oxetane. Subsequent fragmentation experiments generated diagnostic fragments allowing localization positions. Results Intending online hyphenation high-performance liquid chromatography (HPLC)/tandem mass spectrometry (MS/MS), optimized regarding parameters such as time influence trace oxygen well solvents. An separation products means HPLC enabled further characterization chemical structures UV irradiation. To demonstrate applicability developed method, mixture constitutional phospholipid isomers bis(monooleoylglycero) phosphate dioleoyl-phosphatidylglycerol analyzed positions, from extract green alga Chlamydomonas reinhardtii were investigated. Conclusions successful HPLC/MS/MS demonstrated bonds single lipids, mixtures, complex samples. Matrix effects can be reduced structural information, example case isomers, obtained. This opens up avenues even