*2.6. Data Analysis*

The targeted analysis data was analyzed using MultiQuant software (SCIEX, Framingham, MA, USA). The chromatogram peaks were manually curated. The untargeted analysis data was analyzed using MS-DIAL software [17,18] version 2.90 (http://prime.psc.riken.jp/) with the following parameters: retention time start: 0 min; retention time end: 100 min; mass range start: 0 Da; mass range end: 5000 Da; accurate mass tolerance (MS1) tolerance: 0.01 Da; MS2 tolerance: 0.025 Da; maximum charge number: two; smoothing method: linear weighted moving average; smoothing level: 3; minimum peak width: five scans; minimum peak height: 1000; mass slice width: 0.1 Da; sigma window value: 0.5; MS2Dec amplitude cut off: 0; exclude after precursor: true; keep isotope until: 0.5 Da; keep original precursor isotopes: false; exclude after precursor: true; retention time tolerance for identification: 10 min; MS1 for identification: 0.01 Da; accurate mass tolerance (MS2) for identification: 0.05 Da; identification score cut off: 80%; using retention time for scoring: true; relative abundance cut off: 0; top candidate report: true; retention time tolerance for alignment: 0.1 min; MS1 tolerance for alignment: 0.015 Da; peak count filter: 0; adduct ion setting: [M+H]<sup>+</sup>, [M+NH4]<sup>+</sup>, [M+Na]<sup>+</sup>, [M+C2H3N+H]<sup>+</sup>, [M+H-H2O]<sup>+</sup>, [M+C3H8O+H]<sup>+</sup>, [M+C2H3N <sup>+</sup>Na]<sup>+</sup>, [M-C6H10O5+H]<sup>+</sup>, [2M+H]<sup>+</sup>, [2M+NH4]<sup>+</sup>, [2M+Na]<sup>+</sup> in positive ion mode and [M-H]<sup>−</sup>, [M-H2O-H]<sup>−</sup>, [M+CH3COO]<sup>−</sup>, [M-C6H10O5-H]<sup>−</sup>, [2M-H]<sup>−</sup>, and [2M+CH3COO]− in negative ion mode. All lipid classes supported in the MS-DIAL version were used for the targeted lipids. The annotation was performed using the in silico spectral library of lipids, and the false positive annotations were manually curated by mass spectrometry experts. The principal component analysis and circus plots were executed in the R language with the packages of "prcomp" and "OmicCircos". Our untargeted lipidomic

data were discussed in terms of relative quantification, defined by lipidomic standard initiative (https://lipidomics-standards-initiative.org/), among dietary conditions in each tissue. All of the mass spectrometry data are available at RIKEN DROP Met (http://prime.psc.riken.jp/menta.cgi/prime/drop\_ index, ID= DM0030).
