Integrating offline 2D UHPLC-QTOF-MS/MS-based automated analytical platform and UHPLC-QTRAP®-MS: The chemical profiling of modified Gegen Qinlian Decoction

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-09-30 DOI:10.1016/j.jpba.2025.117175

Xinyi Shi , Zhitian Peng , Chongsheng Peng , Xudong Tang , Xiaobo Li

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引用次数: 0

Abstract

The modified Gegen Qinlian Decoction (MGQD) is an upgraded formula of Gegen Qinlian decoction (GQD) with two additional herbs, Euphorbiae Humifusae Herba and Zingiberis Rhizoma Preparatum based on traditional Chinese medicine (TCM) clinical practice. To elucidate the impact of these additions on the chemical composition of the decoction, this study conducted a comprehensive chemical profiling of MGQD. Firstly, the major components in MGQD were quantified, including total saponins, flavonoids, phenolic acids, inorganic elements, carbohydrates, proteins, and free amino acids. These components collectively accounted for 95.06 % of MGQD, with flavonoids being the most abundant category at 30.41 %. Subsequently, an integrated identification strategy was employed by establishing an in-house mass spectral library for MGQD chemical components and incorporating compound clustering analysis through molecular networking of MS² data obtained from offline two-dimensional ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (2D UHPLC-QTOF MS/MS). This approach enabled the identification of 477 structurally diverse compounds in MGQD. Finally, a quantitative analytical method was developed using ultra high performance liquid chromatography coupled with triple quadrupole linear ion trap tandem mass spectrometry (UHPLC-QTRAP®-MS/MS) for the simultaneous determination of 35 selected compounds, which collectively constituted 23.16 % of MGQD. Among these, baicalin was the most abundant (89.4 ± 1.7 μg/mg), followed by puerarin (32.9 ± 0.4 μg/mg) and wogonoside (20.6 ± 0.3 μg/mg). By comparing with the chemical components with that of the original GQD, the potential pharmacodynamic substances of MGQD was inferred, laying the foundation for subsequent research into the pharmacological mechanism. This study provides an effective analytical approach for studying the complex chemical profiles of TCM formulas.

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基于离线二维UHPLC-QTOF-MS/ ms自动分析平台与UHPLC-QTRAP®-MS的集成：改良葛根芩连汤的化学谱分析

改进型葛根芩连汤是在葛根芩连汤的基础上，根据中医临床实践，在原有基础上添加大戟和姜皮两种草药，改进而成。为了阐明这些添加物对汤剂化学成分的影响，本研究对MGQD进行了全面的化学分析。首先，定量分析了其主要成分，包括总皂苷、总黄酮、酚酸、无机元素、碳水化合物、蛋白质和游离氨基酸。这些成分共占MGQD的95.06 %，其中黄酮类化合物含量最高，为30.41 %。随后，采用综合鉴定策略，建立了MGQD化学成分的内部质谱库，并通过分子网络对离线二维超高高效液相色谱-四极杆飞行时间质谱（2D UHPLC-QTOF MS/MS）获得的MS2数据进行化合物聚类分析。该方法鉴定了MGQD中477种结构不同的化合物。最后，建立了超高效液相色谱-三重四极杆线性离子阱串联质谱（UHPLC-QTRAP®-MS/MS）同时测定35个化合物的定量分析方法，共占MGQD的23.16% %。在这其中,黄芩苷是最丰富(89.4 ±1.7  μg /毫克),其次是葛根素(32.9 ±0.4  μg /毫克)和汉黄芩甙(20.6 ±0.3  μg /毫克)。通过与原GQD化学成分的比较，推断出MGQD潜在的药效学物质，为后续药理机制的研究奠定基础。本研究为研究中药复方的复杂化学特征提供了一种有效的分析方法。

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来源期刊

Journal of pharmaceutical and biomedical analysis 医学-分析化学

CiteScore

6.70

自引率

5.90%

发文量

588

审稿时长

37 days

期刊介绍： This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.