Integrated prioritization of candidate quality markers for Smilacis Chinae Rhizoma under the five-principle framework through chemical profiling, effectiveness evaluation, and bionic sensing.

ETHNOPHARMACOLOGICAL RELEVANCE: Smilacis Chinae Rhizoma (SCR), the dried rhizome of Smilax china L., is a traditional Chinese medicinal herb used for disorders associated with inflammation, swelling, pain, and musculoskeletal dysfunction. However, the current pharmacopoeial standard lacks defined assay markers for SCR, limiting the establishment of a more informative quality evaluation system.

AIM OF THE STUDY: This study aimed to prioritize potential quality marker (Q-marker) candidates for SCR under the five-principle framework of measurability, specificity, traceability, effectiveness, and traditional Chinese medicine (TCM) relevance.

MATERIALS AND

METHODS

SCR was chemically profiled by ultrahigh-performance liquid chromatography-quadrupole-Orbitrap mass spectrometry. Specificity and traceability were evaluated through interspecies comparison, herb-distribution analysis, and fresh-to-processed matching. Effectiveness was assessed using a destabilization of the medial meniscus-induced osteoarthritis (OA) model, network pharmacology, molecular docking, interleukin-1β-induced chondrocytes, and molecular dynamics (MD) simulation. TCM relevance was evaluated by electronic tongue and electronic nose analyses.

RESULTS

A total of 226 chemical features/compounds were annotated from SCR, among which 117 were confirmed with authentic standards in the original liquid chromatography-mass spectrometry (LC-MS) workflow, two initially library-assisted candidates were further supported by additional authentic-standard comparison, and the remaining entries were tentatively annotated based on accurate mass, retention time (RT)-associated tandem mass spectrometry (MS/MS) evidence, and diagnostic fragments. Fifty candidates satisfied measurability, specificity, and traceability. SCR alleviated OA-related joint injury in vivo. Network pharmacology prioritized polydatin as a representative effectiveness-related compound, and experimental validation showed that it reduced inflammatory injury and matrix degradation in chondrocytes, partly through inhibition of the TNF/NF-κB axis. MD simulation provided structural support for possible interactions with representative inflammatory network targets. Electronic tongue and electronic nose analyses highlighted the tentatively annotated compounds 4-methoxycinnamic acid and calceolarioside B as taste- and odor-related candidates, respectively.

CONCLUSIONS

By integrating multidimensional evidence under this annotation-confidence framework, 4-methoxycinnamic acid, calceolarioside B, and polydatin were proposed as a potential Q-marker candidate combination for SCR. These findings provide a basis for improving SCR quality control and future assay-marker selection.