Lipid A, the hydrophobic anchor of lipopolysaccharide (LPS) is present in high abundance in Gram-negative bacterial membranes. Individual bacterial species regulate their lipid A structure through a variety of covalent modifications. Analysis of individual structural modifications normally requires analysis by ESI, which poses difficulties due to the hydrophobicity of the lipid A molecule. Surface Acoustic Wave Nebulization (SAWN) produces ions from a flat surface, facilitating lipid A MS and MS~n analysis. Lipid structures are typically assigned manually from the MS~n spectra, which require a great deal of expertise and time. To overcome this low-throughput nature, we demonstrate the ability to assign higher order MS~n spectral results of lipid A using the hierarchical tandem mass spectrometry (HiTMS) algorithm.
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