Abstract
Methodologies for the analysis of intact lipids from fecal matter will facilitate the understanding of gut (patho)physiology, its role(s) in health and disease and help improve patient care. Because lipids are central to intestinal biology, more stable than many metabolites, and more conserved across microbiota than proteins, they will provide information relevant to the microbiome, host, and environment. We developed a fecal extraction method that utilizes two separate, complementary extraction chemistries, dichloromethane and a methyl tert‐butyl ether/hexafluoroisopropanol mixture, alone or with high pressure cycling (PCT), to provide the most robust and comprehensive lipidome coverage. Extracts were assessed by liquid chromatography‐high resolution mass spectrometry profiling with all ion high energy collisional dissociation fragmentation in both positive and negative ionization modes, which enhances unknown lipid characterization. The platform detected >;500 intact lipids, with >;300, spanning 6 LIPIDMAPS categories, identified. This study offers a non‐invasive method to evaluate the GI tract, complement microbiome studies and assess inflammation, nutritional health, and infectious disease in patient vulnerable populations. Research support: K23NR011320 (KEG), U01ES16048 (BSK) and P30DK040561 (WAW).
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