The anti-inflammatory active ingredients of Zhi-Shi-Zhi-Zi-Chi-Tang (ZSZZCT), a traditional
Chinese medicine formula, were predicted and identified using an approach based on
activity index, LC-MS, semi-preparative LC and NMR. Firstly, the whole extract of
ZSZZCT was analyzed using liquid chromatography-quadrupole time of flight-mass spectrometry
(LC-Q-TOF-MS) and liquid chromatography - ion trap mass spectrometry (LC-IT-MS), 79
constituents were detected and 39 constituents were identified unambiguously or tentatively.
Subsequently, the whole extract of the formula was separated into multiple components
and the activity index method was used to calculate index values of the 79 constituents
by integrating the chemical and pharmacological information of multiple components.
Four polymethoxyl flavones were predicted as the major active constituents according
to the activity index values. Furthermore, three polymethoxyl flavones were prepared
using the strategy with semi-preparative LC guided by LC-MS, and their anti-inflammatory
activities were validated. The results show that three polymethoxyl flavones with
higher positive index values, i.e., 3, 5, 6, 7, 8, 3', 4'-heptamethoxyflavone, 3-hydroxynobiletein
and tangeretin had significant anti-inflammatory effects. In conclusion, the predicted
results indicated that the activity index method is feasible for the accurate prediction
of active constituents in TCM formulae.