Profiling of Low-Abundance Branched-Chain Fatty Acids via Radical Directed Dissociation Tandem Mass Spectrometry
Year: 2026
Type: article
Abstract: Branched-chain fatty acids (BCFAs) are key components of the bacterial lipidome, playing a role in regulating membrane fluidity and permeability. In mammals, BCFAs occur at much lower concentrations, and their functions remain largely unexplored. Conventional lipid analysis methods, employing collision-induced dissociation (CID)-tandem mass spectrometry (MS/MS), often fail to locate methyl branching, as fragmentation rarely occurs around the branching site. Here, we introduce a bifunctional derivatization reagent, 1-(8-methoxy-5-quinolinyl) methanamine (MeO-QN), for pinpointing methyl branching in BCFAs with high sensitivity. MeO-QN enhances ionization efficiency of derivatized BCFAs in positive ion mode due to its quinoline moiety and serves as a precursor for radical-directed dissociation (RDD). Upon CID, the quinoline-O radical (QN-O•) is generated, which subsequently induces RDD along the fatty acyl chain and forms a characteristic 28 Da spacing indicative of the branching point. By integrating this MS/MS method with reversed-phase liquid chromatography, we have developed a sensitive analytical workflow, detecting BCFAs at sub-nM levels in mammalian samples. We detected the rarely reported n-5 methyl branched fatty acid (FA 16:0;12Me) in pooled human plasma. We also observed significantly reduced even-chain isobranched fatty acids in breast cancer cells (MDA-MB-468) versus normal breast cells (MCF-10A), suggesting its potential in cancer biomarker discovery.
Institutions: Tsinghua University, Inner Mongolia University
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: bronze
Awards: G2477958377, G6993877796, G8837512327
OpenAlex ID: W7130406145