A mathematical multiple-dimensional strategy for Q-markers identification based on "five principles": Tianshu Capsule for migraine treatment as an example.

Ethnopharmacological Relevance: Quality control is a critical element for Traditional Chinese medicine (TCM). Due to the varied chemical components, mechanisms of action, and pharmacological functions in TCM, ensuring quality is more challenging compared to chemical drugs. Then, the concept of quality markers (Q-markers) was proposed and ideal Q-markers for TCM prescriptions need to compliant with "five principles", including pharmacological effectiveness, specificity, transfer and traceability, measurability, and prescription compatibility.

Aim Of The Study: To establish a mathematical multiple-dimensional "spider-web" strategy and identify the Q-markers of Tianshu capsule (TSC), a Chinese patent medicine for the treatment of migraine, following the "five principles" rules.

Materials And Methods: Q-marker candidates of TSC were firstly screened according to the HPLC fingerprints. Their contents in 10 batches of TSC and stabilities under high temperature, high humidity and in work solutions were determined quantitatively by HPLC-UV (measurability). Their existences in Gastrodiae Rhizoma, Chuanxiong Rhizoma, TSC, rat plasma and brain samples were investigated using HPLC-Q-TOF/MS (transfer and traceability). Their anti-migraine efficacies were evaluated by network pharmacology and mice hot-plate analgesia test; and their relationships with the property (flavor) of Gastrodiae Rhizoma or Chuanxiong Rhizoma were studied by molecular docking (effectiveness). Their contributions were defined based on their herb source according to the compatibility theories of Da Chuan Xiong Fang (compatibility). Their biosynthetic pathways were studied, and their frequencies detected in different plant families were calculated (specificity). Finally, an eight dimensional "spider-web" mode was developed for 10 components, and the regression area (RA) and the coefficient variation (CV) of each candidate were calculated after data normalization.

Results: Ten components including gastrodin, parishin E, chlorogenic acid, ferulic acid, isochlorogenic acid A, senkyunolide I, H, A, Z-ligustilide and levistilide A were selected and evaluated as the Q-marker candidates. The results showed that gastrodin, senkyunolide I, and senkyunolide A had the higher RA and lower CV than other compounds with the established "spider-web" mode, indicating that they could be used as the Q-markers of TSC.

Conclusion: The multi-dimensional "spider-web" mode based on "five principles" was firstly applied to identify the Q-markers of TSC, and it can be used as a practical strategy to discover Q-markers of other compounded prescriptions.