In a benchmark test of LDA versus LipidBlast7 (link), we used data from both the first control experiment and the biological experiment, both acquired on Orbitrap Velos Pro in CID +50% and -50%, and on 4000 QTRAP +45 eV and -45 eV, respectively. For LipidBlast evaluation, we used the recommended MSPepSearchGUI (http://peptide.nist.gov/software/ms_pep_search_gui/MSPepSearch.html). The same m/z tolerances were applied in both LDA and LipidBlast. The specificity and sensitivity of LipidBlast depend on a so called matching factor22 (link), a value ranging from 0-999. Using the default setting of 450 for the matching factor, many lipid standards in control experiment 1 were not detected. Consequently, the matching factor was lowered to 10, in which case LipidBlast detected almost all of the lipid standards in negative ion mode. Further reduction did not improve the sensitivity of LipidBlast. In positive ion mode, irrespective of the matching factor setting, LipidBlast was not able to identify as many lipid molecular species as was LDA. Details about the LipidBlast parameters are given in Supplementary Note 2. In this benchmark, we used only lipid subclasses/adducts that both LDA and LipidBlast are able to detect. Correct assignment of lipid species and lipid molecular species identified in liver lipidomes was verified by manual inspection of the spectra, and by aligning them with the respective retention time data23 (link).