Fig. 1.

Flowchart of the strategy used to conduct a widely targeted metabolomics analysis. Automated FIA-TQMS was applied to a total of 967 authentic compounds to obtain an ESI-MS/MS data set consisting of 61,920 spectra for 860 authentic compounds. The remaining 107 compounds did not show reproducible spectra with FIA-TQMS. The MRM data set consists of optimized analytical conditions for 497 compounds, determined automatically, based on the ESI-MS/MS data set. UPLC-TQMS analysis was applied to these 497 compounds using the optimized UPLC and MRM conditions. Approximately 80% (378) of the compounds were successfully detected, to give a retention time index data set. The conditions that were established, as described above, were used in combination with an ALHS to conduct a widely targeted metabolomics study of 14 accessions representing nine plant species from three families, and the metabolite accumulation patterns were determined. All large-scale data sets mentioned here are freely available at the PRIMe website.