Peng Zhitian, Peng Chongsheng, Peng Ying, Li Xiaobo
School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
Phytochem Anal. 2025 Aug;36(6):1803-1819. doi: 10.1002/pca.3547. Epub 2025 Jun 2.
An integrated strategy was utilized in this study to comprehensively characterize the chemical composition of Gegen-Qinlian Decoction (GQD).
The major components of GQD (total flavonoids, total saponins, total alkaloids, carbohydrates, and proteins) were determined firstly using colorimetric methods, while free amino acids and trace elements were analyzed by automatic amino acid analyzer and inductively coupled plasma-mass spectrometry (ICP-MS), respectively. After that, the small-molecule components of GQD were characterized using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS). Furthermore, a quantitative method was established for 26 representative compounds determination simultaneously using ultra-high performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UHPLC-QTRAP-MS/MS) in multiple reaction monitoring (MRM) mode.
The accurate content and proportion of the major constituents in GQD were determined for the first time. A total of 230 compounds were characterized from GQD, of which four undescribed alkaloids were identified initially by their tandem mass data analyses based on mass spectral fragmentation pathways of isoquinoline alkaloids. A quantitative method was established to determine 26 representative compounds, and validated through linearity, precision, repeatability, stability, and recovery. The total content of the 26 compounds accounted for 27.81%, of which baicalin being the most abundant was up to 107.26 ± 1 mg/g.
The present study achieved a comprehensive characterization of constituents in GQD, laying the foundation for elucidating the active components, mechanism, and the quality control of GQD, offering a valuable analytical framework for research on other traditional Chinese medicine (TCM) formulas.
本研究采用综合策略全面表征葛根芩连汤（GQD）的化学成分。
首先采用比色法测定GQD的主要成分（总黄酮、总皂苷、总生物碱、碳水化合物和蛋白质），同时分别用自动氨基酸分析仪和电感耦合等离子体质谱仪（ICP-MS）分析游离氨基酸和微量元素。之后，采用超高效液相色谱-四极杆飞行时间串联质谱（UHPLC-QTOF-MS/MS）对GQD的小分子成分进行表征。此外，建立了一种定量方法，采用超高效液相色谱-三重四极杆线性离子阱串联质谱（UHPLC-QTRAP-MS/MS）在多反应监测（MRM）模式下同时测定26种代表性化合物。
首次确定了GQD中主要成分的准确含量和比例。从GQD中鉴定出230种化合物，其中根据异喹啉生物碱的质谱裂解途径，通过串联质谱数据分析初步鉴定出4种未描述的生物碱。建立了测定26种代表性化合物的定量方法，并通过线性、精密度、重复性、稳定性和回收率进行了验证。这26种化合物的总含量占27.81%，其中含量最高的黄芩苷达107.26±1mg/g。
本研究实现了对GQD成分的全面表征，为阐明GQD的活性成分、作用机制和质量控制奠定了基础，为其他中药方剂的研究提供了有价值的分析框架。
