目的 筛选柴胡-黄芩（柴黄）药对解热作用质量标志物（quality marker，Q-Marker），并建立Q-Marker的UPLC含量测定方法。方法 采用均匀设计法建立柴黄药对配伍差异样品，利用超高效液相色谱-四级杆-静电场轨道阱质谱（UPLC- Q-Exactive Orbitrap-MS）技术建立柴黄药对煎液指纹图谱，采用Thermo Scientific Hypersil Gold C₁₈色谱柱（150 mm×2.1 mm，3 µm），以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱，体积流量0.3 mL/min，并采用电喷雾电离，正离子扫描模式下进行质谱数据采集。采用新西兰兔耳缘静脉脂多糖（lipopolysaccharides，LPS）注射的方法建立少阳证高热模型，以体温变化值作为解热药效指标，采用灰色关联度分析法建立指纹图谱中共有峰与解热作用的相关关系，筛选出关联性较大的成分作为柴黄药对解热作用的主要Q-Marker，采用UPLC建立Q-Marker含量测定方法。结果 筛选出黄芩苷、汉黄芩苷、千层纸素A-7-O-葡萄糖醛酸苷、去甲汉黄芩素-7-O-葡萄糖醛酸苷、黄芩素、汉黄芩素、柴胡皂苷A、柴胡皂苷C作为柴黄药对解热Q-Marker，并建立了含量测定方法。结论 基于UPLC-Q-Exactive Orbitrap-MS技术的谱-效相关研究策略可以快速分析及初步确定柴黄药对解热Q-Marker，在此基础上建立其UPLC含量测定方法，对优化柴黄类方制剂的质量控制方法具有重要的参考价值。

Objective To screen the quality markers of Chaihu (Bupleuri Radix) and Huangqin (Scutellariae Radix) drug pair (BSDP) decoction, and establish the UPLC content determination method of quality markers (Q-Marker). Methods UPLC-Q-Exactive Orbitrap-MS was used to establish the fingerprint of BSDP decoction. The analysis was performed on a Thermo Scientific Hypersil Gold C₁₈ column (150 mm×2.1 mm, 3 μm) by gradient elution, and the mobile phase consisted of acetonitrile and 0.1% formic acid solution. Electron spray ionization was used, via positive ions scanning. The New Zealand rabbit model with Shaoyang fever syndrome was established by LPS injection via ear vein. Taking temperature change value as antipyretic effect index, gray correlation analysis was used to study the correlation between the common peaks in the fingerprints and the antipyretic effect to screen out the components with greater correlation as quality markers for the antipyretic effect of BSDP decoction, and the contents determination method for quality markers was established using UPLC. Results Eight quality markers related to the antipyretic effect of BSDP decoction were screened out: baicalin, wogonoside, oroxyloside, glychionide A, baicalein, wogonin, saikosaponin A and saikosaponin C, and the content determination method of them was established. Conclusion The “spectrum-effect” correlation research strategy based on UPLC-Q-Exactive Orbitrap-MS technology is an effective method for rapid analysis and preliminary determination of antipyretic quality markers of BSDP. On the basis, the UPLC method established for content determination of the eight quality markers is reliable and accurate. This study has important reference value for the in-depth research on quality control of the compound preparation containing BSDP.

R283.6

山东省自然科学基金项目(ZR2017PH068);山东省重点研发计划项目(2018GSF119019);山东省中医药科技项目(2020Z08)