Lian Zhu,Shengjun Ma,Kailin Li,Pei Xiong,Shihan Qin,Wei Cai
Arnebiae Radix(dried root ofArnebia euchroma(Royle) Johnst.) has been used in traditional Chinese medicine (TCM) to treat macular eruptions, measles, sore throat, carbuncles, burns, skin ulcers, and inflammation. Previous studies have shown that shikonins and shikonofurans are two of their main bioactive ingredients. However, systematic investigations of their constituents have rarely been conducted. It is necessary to establish a rapid and effective method to identify the chemical constituents ofArnebiae Radix. This will help to further improve the effective resource utilization rate of this plant. In this study, a rapid and effective UHPLC-Q-Exactive Orbitrap mass spectrometry method was established to simultaneously analyze chemical ingredients inArnebiae Radixwithin a short period of time. Based on the results of a full scan MS, the MS2database (mzVault and mzCloud), the diagnostic fragment ions, the retention time, and the bibliography, a total of 188 compounds were identified, with 114 of those being reported fromArnebiae Radixfor the first time. The results of this study lay the foundation for obtaining a thorough understanding of the active ingredients inArnebiae Radixand its quality control. This method may be widely used for the chemical characterization of different samples.Keywords:Arnebiae Radix;identification;chemical constituents;UHPLC-Q-Exactive Orbitrap MS
Arnebiae Radix (dried root of Arnebia euchroma (Royle) Johnst.) is a traditional Chinese medicine (TCM) used to treat macular eruptions, measles, sore throat, carbuncles, burns, skin ulcers, and inflammations. The Arnebiae Radix extract can exert anti-breast cancer effects through various mechanisms of action. This study aimed to rapidly screen potential estrogen receptor (estrogen receptor α and estrogen receptor β) ligands from the Arnebiae Radix extract. In this study, an analytical method based on affinity ultrafiltration coupled with UHPLC-Q-Exactive Orbitrap mass spectrometry was established for rapidly screening and identifying estrogen receptor ligands. Then, bindings of the components to the active site of estrogen receptor (estrogen receptor α and estrogen receptor β) were investigated via molecular docking. Moreover, surface plasmon resonance (SPR) experiments with six compounds were performed to verify the affinity. As a result, a total of 21 ligands were screened from Arnebiae Radix using affinity ultrafiltration. Among them, 14 and 10 compounds from Arnebiae Radix showed affinity with estrogen receptor α and estrogen receptor β, respectively. All of those ligands could have a good affinity for the multiple amino acid residues of the estrogen receptor based on molecular docking. In addition, six compounds display the great affinity by SPR. The method established in the study could be used to rapidly screen estrogen receptor ligands in Traditional Chinese medicine. The results demonstrated that the affinity ultrafiltration–UHPLC-Q-Exactive Orbitrap mass spectrometry method not only aids in the interpretation of the potential bioactive components and possible mechanisms of action of Arnebiae Radix but also provides a further effective basis for the quality control of this valuable herb medicine.
Background The “one-to-multiple” phenomenon and counterfeiting are prevalent in herbal medicines. Accurately identifying these issues in both crude herbs and commercial formulated products is crucial for ensuring drug safety and efficacy. However, this task remains inherently challenging due to the complex nature of the matrices involved. Purpose This study aims to develop methodologies for the precise authentication of multi-sourced herbal medicines and their counterfeits, and to trace constitute species in commercial products. Methods This paper proposes an integrated strategy termed “Core-Multiplex-Micro (CMM) component profiling for precision discrimination amid multi-sources and adulterants”, using Arnebiae Radix as a case study. The strategy begins with the comparative analysis of primary component, shikonin derivatives, across two genuine species and three adulterants, emplying single standard to determine multi-components (SSDMC) approach. It then expands to include a broader range of mid- to low-abundance metabolites through fingerprinting-based classification. Subsequently, trace-level components are explored using LTQ-Orbitrap MS combined with metabolomics to authenticate the five species. Robust chemical markers are identified, validated, and translated to a cost-effective QDa MS platform for targeted detection. This tiered methodology was applied to authenticate 15 batches of commercially available raw materials and 10 batches of formulations; Results The study successfully distinguished between two authentic Arnebiae Radix species and three adulterants simultaneously for the first time. Through cross-validation, nine diagnostic chemical markers were identified and effectively transferred from expensive LTQ-Orbitrap MS platform to more cost-effective QDa MS system for practical use in commercial product traceability. The research revealed that 60% raw Arnebiae Radix materials were authentic, while 30% formulations matched their labeled species composition. Conclusion This study sets a precedent for precise traceability for “one-to-multiple” and counterfeit products: tracing them from raw herbs to final formulations.
危险品化学品经营许可证（带存储）