Wang Long, Wu Weigang, Li Guoxiang, Chen Haiyang, Fan Yinyin, Chen Wei, Zhou Guifang, Li Wenlong
College of Pharmaceutical Engineering of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
Tianjin Key Laboratory of Intelligent and Green Pharmaceuticals for Traditional Chinese Medicine, Tianjin, China.
Phytochem Anal. 2025 Apr;36(3):832-845. doi: 10.1002/pca.3471. Epub 2024 Nov 4.
The inherent complexity of traditional Chinese medicine (TCM) poses significant challenges in directly correlating quality evaluation with clinical efficacy. Banxia-Houpo Decoction (BHD), a classical TCM formula, has demonstrated efficacy in treating globus hystericus. However, the intricate composition of BHD, which contains both volatile and non-volatile active components, complicates efforts to ensure its consistent quality and clinical effectiveness.
The aim of this study was to introduce an integrated approach that combines non-targeted multicomponent analysis, network pharmacology, and multivariate chemometrics to identify quality markers for the effective quality control of BHD.
First, a nontargeted high-definition MSE method based on ultraperformance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UHPLC-QTOF-MS) was developed for the comprehensive multi-component characterization of BHD. Next, the quality markers of nonvolatile compounds in BHD were identified through network pharmacology analysis. Subsequently, volatile organic compounds (VOCs) in BHD samples were analyzed via headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS). Finally, the orthogonal partial least squares discriminant analysis (OPLS-DA) model was applied to screen for potential markers.
Based on in-house library-driven automated peak annotation and comparison with 25 reference compounds, 128 components were identified for the first time. Additionally, honokiol, magnolol, magnoflorine, 6-gingerol, rosmarinic acid, and adenosine were preliminarily identified as potential quality markers for BHD through network pharmacology analysis. By employing two complementary techniques, HS-SPME-GC-MS and HS-GC-IMS, a total of 145 volatile compounds was identified in the BHD samples. Four potential differential VOCs in the BHD samples were further identified based on the variable importance in projection (VIP ≥ 1.5) using HS-GC-IMS combined with chemometric analysis.
In conclusion, this study not only contributes to establishing quality standards for BHD but also offers new insights into quality assessment and identification in the development of classical formulations enriched with volatile components.
中药固有的复杂性给将质量评价与临床疗效直接关联带来了重大挑战。半夏厚朴汤（BHD）是一种经典的中药方剂，已证明在治疗癔球症方面有效。然而，BHD的成分复杂，既含有挥发性活性成分又含有非挥发性活性成分，这使得确保其质量和临床疗效的一致性变得复杂。
本研究的目的是引入一种综合方法，将非靶向多成分分析、网络药理学和多元化学计量学相结合，以确定用于半夏厚朴汤有效质量控制的质量标志物。
首先，开发了一种基于超高效液相色谱 - 四极杆飞行时间质谱（UHPLC-QTOF-MS）的非靶向高清MSE方法，用于半夏厚朴汤的综合多成分表征。接下来，通过网络药理学分析确定半夏厚朴汤中非挥发性化合物的质量标志物。随后，通过顶空固相微萃取 - 气相色谱 - 质谱（HS-SPME-GC-MS）和顶空气相色谱 - 离子迁移谱（HS-GC-IMS）分析半夏厚朴汤样品中的挥发性有机化合物（VOCs）。最后，应用正交偏最小二乘法判别分析（OPLS-DA）模型筛选潜在标志物。
基于内部库驱动的自动峰注释并与25种参考化合物进行比较，首次鉴定出128种成分。此外，通过网络药理学分析，厚朴酚、厚朴碱、木兰碱、6-姜酚、迷迭香酸和腺苷被初步确定为半夏厚朴汤的潜在质量标志物。通过采用HS-SPME-GC-MS和HS-GC-IMS这两种互补技术，在半夏厚朴汤样品中总共鉴定出145种挥发性化合物。基于HS-GC-IMS结合化学计量分析的投影变量重要性（VIP≥1.5），进一步鉴定出半夏厚朴汤样品中的4种潜在差异VOCs。
总之，本研究不仅有助于建立半夏厚朴汤的质量标准，还为富含挥发性成分的经典方剂开发中的质量评估和鉴定提供了新的见解。
