Abstract:

Traditional Chinese medicine （TCM）， a treasured cultural heritage of the Chinese nation， plays a significant role in treating various diseases. However， the extreme complexity of its chemical components makes the comprehensive elucidation of the material basis and pharmacological mechanisms a major challenge in modern TCM research. To deeply investigate the material basis of TCM and interpret its holistic regulation mechanism involving multiple components and targets， Liang Xinmiao and his colleagues proposed the concept of herbal materiomics， which focuses on two main research directions： the material basis （including qualitative/quantitative analysis and isolation of active compounds） and the functional mechanisms （in-depth study of biological effects and modes of action）. In herbalome research， chromatographic analysis and preparative purification are powerful tools for deciphering the composition of complex TCM systems， with the development of novel chromatographic separation materials being a key driver of progress. This review systematically summarizes recent advances in the development of novel high performance chromatographic separation materials since 2020 and their applications in herbalome research. It focuses on three chromatographic modes： reversed-phase liquid chromatography （RPLC）， hydrophilic interaction liquid chromatography （HILIC）， and supercritical fluid chromatography （SFC）. The structural and performance characteristics of these novel materials are highlighted， along with their applications in the analysis and preparative purification of key active components such as alkaloids， flavonoids， saponins and terpenoids. In RPLC， the separation selectivity and retention capacity of the stationary phases for compounds such as alkaloids and flavonoids have been effectively enhanced through two primary strategies： first， by incorporating polar groups such as fluorine atoms， amides， carboxyl groups into the traditional C18/C8/phenyl skeletons； second， by utilizing natural product derivatives like cardanol and rosin as functional ligands. Furthermore， this approach has led to the creation of novel materials possessing orthogonal separation characteristics. In HILIC， various functionalized stationary phases have been developed， including β-cyclodextrin， nitrogen-containing heterocycles， core-shell structured amides， and surface-polymerized phases. These phases significantly improve the retention and separation efficiency of strongly polar compounds （e.g.， saponins， oligosaccharides） by providing abundant hydrogen bonding， electrostatic， and π-π interactions. In SFC， the development of novel stationary phases extends from modified functional skeletons （e.g.， phenyl） to zwitterionic and nitrogen-containing heterocyclic hydrophilic stationary phases. Additionally， mixed-mode stationary phases have been developed to effectively ameliorate the peak tailing problems associated with basic compounds. Concurrently， the emergence of small-particle-size ordered mesoporous silica materials enables ultra-high column efficiency and rapid separation of components such as tanshinones. This review further illustrates the successful application of these novel materials in both high-performance analytical and preparative chromatography， using four important classes of TCM active ingredients—alkaloids， flavonoids， saponins， and terpenoids—as examples. These novel materials afford significant chromatographic improvements—notably in separation selectivity， peak symmetry， analysis speed， and the resolution of previously difficult-to-separate isomers. Critically， the integration of these advanced materials has facilitated the construction of highly orthogonal multidimensional chromatographic systems （e.g.， RPLC×RPLC， HILIC×RPLC）， thereby providing the capacity for in-depth profiling and identification of dozens to hundreds of components from complex TCM extracts. In the field of preparative chromatography， these materials enable efficient and large-scale purification of target monomers， including trace active ingredients， thereby significantly enriching compound libraries for herbalome research. This includes the implementation of sophisticated multi-dimensional and multi-mode preparative strategies， which have facilitated the discovery of novel compounds and unprecedented skeletons. Finally， the article concludes by outlining future development directions of chromatographic materials， emphasizing the need to enhance separation selectivity and orthogonality， improve separation efficiency and preparative loading capacity， and develop versatile stationary phases. These advancements are crucial to better support the in-depth development of herbalome research. This review aims to provide a material foundation and technical reference for resolving the material basis of complex TCM systems.

Key words: chromatographic separation materials, herbalome, separation and analysis, preparative chromatography