目的 建立薤白特征图谱分析方法，并结合化学计量学和一测多评法（quantitative analysis of multi-components with a single-marker，QAMS）开展薤白质量控制研究。方法 采用HPLC方法，Agilent Eclipse Plus C₁₈色谱柱（250 mm×4.6 mm，5μm），以甲醇-超纯水溶液为流动相梯度洗脱，体积流量1.0 mL/min，柱温30℃，检测波长261 nm，进样量20 μL，建立薤白特征图谱，接着采用聚类分析（clustering analysis，CA）、主成分分析（principal component analysis，PCA）、正交偏最小二乘法-判别分析（orthogonal partial least square method-discriminant analysis，OPLS-DA）对不同批次薤白饮片进行化学模式识别分析，实现数据集的可视化。同时以腺苷为内参物建立QAMS法，建立内参物与胞苷、尿苷、鸟苷的相对校正因子，采用外标法（external standard method，ESM）与QAMS法分别测定12批次薤白样品中胞苷、尿苷、鸟苷、腺苷4种核苷类成分的含量，以验证QAMS法结果的可靠性。结果 建立的HPLC特征图谱方法较全面地反映了薤白药材的化学成分，分析方法专属性强，准确可靠；化学计量学分析结果基本一致；并通过OPLS-DA找出了3种可能的标志性成分；QAMS法与外标法测定结果之间无显著差异。结论 建立的薤白特征图谱分析方法、化学计量学方法、多指标成分QAMS法可为薤白和其他植物药的质量控制与评价提供参考。

Objective To establish an analytical method used for quality control of Xiebai (Allii Macrostemonis Bulbus, AMB) employing HPLC characteristics spectrum combined with stoichiometry and quantitative analysis of multi-components with a single-marker method. Methods HPLC was performed on Agilent 1200 HPLC system equipped with DAD detector and Agilent Eclipse Plus C₁₈ column (250 mm×4.6mm, 5µm), using methanol-ultrapure water solution as mobile phase employing gradient elution procedure with the flow rate of 1.0 mL/min, column temperature of 30℃, detection wavelength of 261 nm, and injection volume of 20 μL. Cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to identify chemical patterns of different batches of AMB at a visualization analysis level. The QAMS method was also adopted to evaluate the main components contents including adenosine, cytidine, uridine, and guanosine in AMB samples with adenosine as the internal reference, and the relative correction factors of the internal reference and cytidine, uridine and guanosine were established subsequently. Finally, to confirm validity of QAMS method, the comparison of the contents data analyzed by QAMS method and external standard method (ESM) was also carried out. Results The established HPLC characteristics spectrum method can fully reflect the chemical composition of AMB. The analysis method showed specific, accurate and reliable characteristics and the relative correction factors have good repeatability. Moreover, the results of chemometric analysis were in accord with data calculated from methods mentioned above. In addition, three possible characteristic components were identified by OPLS-DA and there was no significant difference found between QAMS and ESM, which can provide a reference for quality evaluation of AMB. Conclusion The established HPLC fingerprint spectrum method, stoichiometry method and multi-index components analytical method with QAMS can provide a reference for quality control and evaluation of AMB and other botanic drugs.

R286.2

国家自然科学基金资助项目（81973211/C0033375）；湖南省中医药科研计划项目［D2022139（A2022005-17）］；湖南省大学生创新创业训练计划项目（湘教通［2022］174号-2918）；湖南中医药大学研究生创新课题（校行研字［2022］20号-2022CX87）；湖南中医药大学“十四五”重点学科-生物工程学科（校行发规字[2023]2号）