A strategy based on chemical markers’ fishing and knockout has been proposed for holistic activity and
interaction evaluation of the bioactive components in herbal medicines (HMs). It was devised to screen
bioactive-compound group that represents the efficacy of HM, estimate the bioactivity contribution of
each component and elucidate the interactions of multi-components. This strategy was accomplished
through the following steps: (1) screen out the chemical markers (target peaks) in a HM fingerprint
using online two-dimensional turbulent flow chromatography/liquid chromatography–mass spectrometry
technique, (2) fish target peaks and knockout any interested peak, and (3) evaluate the bioactivities
of fishing and knockout portions. After comparison of the bioactivities of samples containing different
target peaks, the efficacy of target-peak group, bioactivity contribution of each compound, and the
interactions of multi-components are elucidated. Using Acetylcholinesterase (AChE) and Bulbs of Lycoris
radiata (L. Herit.) Herb