A Five-Dimensional "Spider-Web" Strategy for Quality Markers Identification of Traditional Chinese Medicine Compound...

Given the complex composition of traditional Chinese medicine (TCM) formulas, quality evaluation cannot rely on a single component or a limited few. Therefore, a five-dimensional quality marker (Q-marker) identification system encompassing "effectiveness, specificity, transferability, measurability, and formula compatibility" was developed and subsequently applied to Fufang Huangqi Jianpi oral liquid (FFHQJPOL) as an example. Using UHPLC-Q-TOF/MS, 62 compounds were identified in the preparation. The measurability dimension was constructed based on 17 major constituents derived from 23 common peaks in the chromatographic fingerprints of 10 batches. Serum pharmacochemistry via UHPLC-Q-TOF/MS revealed 32 prototype components absorbed into the bloodstream, thereby establishing the transferability dimension. Network pharmacology approaches were applied to construct a "component-target-disease-pathway" network, defining the effectiveness dimension. A topological network of compounds and herbs was established to depict the distribution pattern of the candidate compounds across various herbs, thereby revealing specificity. The compound compatibility dimension was established based on the function and proportional weight of its active ingredients in the formula. Ultimately, through regression area (RA) and coefficient of variation (CV) calculation, 3-hydroxy-9,10-dimethoxypterocarpan, ononin, zizyphusine, calycosin-7-O-glucoside, and sinapine were initially identified as the key Q-markers. It is worth mentioning that astragaloside IV and astragaloside II were additionally incorporated into the Q-markers system based on UHPLC-Q-TOF/MS analysis and their known pharmacological activities, ultimately establishing seven key Q-markers for FFHQJPOL. This strategy provided a multi‑dimensional integrated paradigm for Q‑markers discovery in TCM compound preparations, facilitating the transition from single‑index to system‑integrated quality control.