Quan Jian-Ye, Fan Bin, Liu An, Sun Jian, Chen Peng, Wang Chun-Guo, Zhao Yong-Lie, Zhang Cun, Deng Xin-Qi, Jing Zhi-Wei
Experimental Research Center, China Academy of Chinese Medical Sciences Beijing 100700, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2023 Jan;48(1):71-81. doi: 10.19540/j.cnki.cjcmm.20220705.305.
Wuzhuyu Decoction, the classical formula recorded in the Treatise on Febrile Diseases(Shang Han Lun), has been included in the Catalogue of Ancient Classic Prescriptions(the First Batch). Consisting of Euodiae Fructus, Ginseng Radix et Rhizoma, Zingiberis Rhizoma Recens, and Jujubae Fructus, it is effective in warming the middle, tonifying deficiency, dispelling cold, and descending adverse Qi, and is widely applied clinically with remarkable efficacies. For a classical formula, the chemical composition is the material basis and an important premise for quantity value transfer. This study aimed to establish a rapid identification method of chemical components in Wuzhuyu Decoction by high-resolution mass spectrometry(HR-MS) and molecular network. AQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was used for sample separation, and acetonitrile-0.1% formic acid in water was used as mobile phases for gradient elution. Q-Exactive Orbitrap MS data were collected in positive and negative ion modes, and GNPS molecular network was plotted according to the similarity of MS/MS fragmentation modes. Cytoscape 3.6.1 was used to screen molecular clusters with similar structures. Finally, the chemical components of Wuzhuyu Decoction were rapidly identified according to the controls, as well as the information of retention time, accurate relative molecular weight of HR-MS, and MS/MS multistage fragments. A total of 105 chemical components were identified in Wuzhuyu Decoction. This study can provide data for the follow-up quality control, standard substance research, and pharmacodynamic material research on Wuzhuyu Decoction, as well as references for the rapid qualitative analysis of the chemical components of Chinese medicine.
吴茱萸汤是《伤寒论》中记载的经典方剂，已被列入《古代经典名方目录（第一批）》。该方由吴茱萸、人参、生姜、大枣组成，具有温中补虚、散寒降逆之功效，临床应用广泛，疗效显著。对于经典方剂而言，化学成分是其物质基础，也是量值传递的重要前提。本研究旨在通过高分辨质谱（HR-MS）和分子网络建立吴茱萸汤化学成分的快速鉴定方法。采用ACQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm，1.7 μm）进行样品分离，以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱。在正、负离子模式下采集Q-Exactive Orbitrap MS数据，并根据MS/MS碎片模式的相似性绘制GNPS分子网络。利用Cytoscape 3.6.1筛选结构相似的分子簇。最后，根据对照品以及保留时间、HR-MS精确相对分子量和MS/MS多级碎片信息，快速鉴定吴茱萸汤的化学成分。共鉴定出吴茱萸汤中的105种化学成分。本研究可为吴茱萸汤后续的质量控制、标准物质研究及药效物质研究提供数据，也可为中药化学成分的快速定性分析提供参考。
