目的 建立经典名方槐花散的指纹图谱，并通过化学成分辨识和多组分定量整合分析方法，对其进行“质”与“量”的综合评控。方法 采用超高效液相-三重四级杆液质（UPLC-triple quadrupole liquid chromatography-mass spectrometry，UPLC-QQQ-MS/MS）联用技术，首次以一级全扫（Full-MS）和多反应监测（multiple reaction monitoring，MRM）2种采集模式相结合的方式建立槐花散UPLC-QQQ-MS/MS指纹图谱，夹角余弦法进行相似度计算。以方中芦丁、柚皮苷、槲皮苷、新橙皮苷、木犀草素、槲皮素、山柰酚和胡薄荷酮8种质量标志物作为评价指标，进行含量测定，筛选出最优产地药材组合处方；结合超高效液相-高分辨液质联用技术（UPLC-Q-Orbitrap HRMS）对其化学成分进行系统辨识。采用Thermo Fisher Accucore^TM C₁₈色谱柱（100 mm×3.0 mm，2.6 μm）；乙腈-0.1%甲酸水溶液为流动相，进行梯度洗脱。三重四级杆质谱扫描方式为Full-MS与MRM相结合；高分辨质谱扫描方式为全扫描/数据依赖二级扫描（Full MS/dd-MS²），2种质谱均为电喷雾离子源（ESI）。结果 建立了不同产地4味药材40种随机组合的经典名方槐花散的UPLC-QQQ-MS/MS指纹图谱，在Full-MS扫描模式下标定了21个共有峰，相似度在0.745～0.977，通过对照品结合MRM扫描模式对其中芦丁等8种化学成分进行了特征峰指认。定量测定结果显示槐花散S1～S40供试品中7种黄酮类成分质量分数总和在13.238～43.461 mg/g，相差较为明显；其中S14中总黄酮含量高于S33逾3倍；胡薄荷酮质量分数在S15中最高，达2.528 mg/g。最终确定S20为最优组合处方，其7种黄酮类成分质量分数总和达43.018 mg/g，胡薄荷酮质量分数为2.498 mg/g。采用UPLC-Q-Orbitrap HRMS技术对最优处方组合的供试品S20进行化学成分辨识，共鉴定出90个化合物，包括黄酮类35个、氨基酸类5个、生物碱类3个、糖类2个、脂肪酸8个、萜类7个、有机酸类3个、香豆素类2个、酚酸类4个、其他类21个。结论 UPLC-QQQ-MS/MS与UPLC-Q-Orbitrap HRMS 2种质谱技术相结合建立了不同产地4味药材40种随机组合的槐花散指纹图谱，并对其化学成分进行了系统定性定量分析，具有客观、高效、准确等优势，为挖掘槐花散药效物质基础、开发中药复方制剂的质量控制手段提供有力参考和科学依据。

Objective To establish the fingerprint of classical famous formula Huaihua Powders (槐花散), and evaluate its quality and quantity by chemical composition identification and multi-component quantitative integration analysis. Methods Using UPLC-triple quadrupole liquid chromatography-mass spectrometry (QQQ-MS/MS) coupling technology, the UPLC-QQQ-MS/MS fingerprint of Huaihua Powders was established by the combination of Full-MS and multiple reaction monitoring (MRM) for the first time, and the similarity was evaluated by the included Angle and cosine method. Eight quality markers of rutin, naringin, quercitrin, neohesperidin, luteolin, quercetin, kaempferol and humenthone were used as evaluation indexes to determine the content and select the optimal combination prescription. The chemical composition was identified by UPLC-Q-Orbitrap HRMS. The chromatography was performed on Thermo Fisher Accucore^TM C₁₈ column (100 mm×3.0 mm, 2.6 μm). Acetonitrile-0.1% formic acid aqueous solution was used as mobile phase for gradient elution. The scanning mode of triple four-bar mass spectrometry was the combination of Full-MS and MRM. The high resolution mass spectrometry scanning method was Full scan/data-dependent secondary scan (Full MS /dd-MS2), and both germplasm spectra were electrospray ion sources (ESI). Results The UPLC-QQQ-MS/MS fingerprint chromatograms of the classic prescription Huaihua Powders with 40 random combinations of four medicinal materials from different origins were established. A total of 21 common peaks were identified in Full-MS scanning mode, with the similarity ranging from 0.745 to 0.977. The characteristic peaks of eight chemical components, including rutin, were identified by reference and MRM scanning mode. The content determination results showed that the total content of seven flavonoids in Huaihua Powders S1—S40 was 13.238—43.461 mg/g, and the difference was obvious. The content of total flavonoids in S14 was three times higher than that in S33. The content of humenthone was the highest in S15, reaching 2.528 mg/g. S20 was identified as the best combination prescription, the total content of seven flavonoids was 43.018 mg/g, and the content of humenthone was 2.498 mg/g. UPLC-Q-Orbitrap HRMS technique was used to identify the chemical components of S20 with optimal prescription combination. A total of 90 compounds were identified, including 35 flavonoids, five amino acids, three alkaloids, two sugars, eight fatty acids, seven terpenoids, three organic acids, two coumarins, four phenolic acids and 21 other compounds. Conclusion The fingerprint chromatograms of 40 random combinations of four medicinal materials from different origins in Huaihua Powders were established by combining two mass spectrometry techniques, UPLC-QQQ-MS/MS and UPLC-Q-Orbitrap HRMS, and a systematic qualitative and quantitative analysis of their chemical components was conducted, which was objective, efficient and accurate. It provides a strong reference and scientific basis for exploring the pharmacodynamic material basis of Huaihua Powders and developing the quality control means of TCM compound preparations.

R283.6

国家社会科学基金资助项目（ CGA220304）；四川省自然科学基金青年基金（2025ZNSFSC1823）；成都中医药大学“杏林学者”学科人才科研提升计划苗圃人才专项（MPRC2022017）