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Ultra-performance liquid chromatographytandem mass spectrometry (UPLC-MS/MS) enables precise quantification of structurally diverse BA species (175, 176), facilitating detailed characterization of the bile acid-ome. The emerging field of reverse metabolomics further accelerates the discovery of novel bioactive BA derivatives, including previously unannotated microbial metabolites, directly detected in human-derived biospecimens ( Beyond bulk analyses, single-cell technologies such as single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics/metabolomics are now providing unprecedented resolution of cellular heterogeneity and spatio-temporal BA signaling within the TIME and associated mucosal tissues (177)