The comprehensive 2D NQO1 biochromatography system was aboard on an Agilent 1200 series high-performance liquid chromatography system combined with time-of-flight mass spectrometer (TOFMS), controlled by Agilent MassHunter Workstation (Agilent Technologies, Palo Alto, CA, USA). With ammonia acetate as the mobile phase, the NQO1 column was used as the first-dimensional column. Besides, an Agilent Poroshell 120 EC-C18 column was used as the second dimension, with acetonitrile and 0.1% formic acid as the mobile phase. The relevant operating details of the comprehensive 2D NQO1 column/C18 column/TOFMS system could be referred to in the literature [22 (link), 23 (link)]. The system was equilibrated for 30 min, and 3 µL of the Scutellaria baicalensis extract was injected into the system. In order to obtain the comprehensive 2D NQO1 biochromatographic analysis results, system collects at least two fractions across a retention peak. Therefore, the collection period of each round was set to 2.5 min. Subsequently, the machine collected the information on the stable retention behavior and molecular structure of components in extract of Scutellaria baicalensis. Relevant data were analyzed for baseline correction and 2D mapping of retention behavior.