目的 建立连翘Forsythiae Fructus的UPLC指纹图谱及多成分含量测定方法，并结合化学计量学方法寻找不同产地连翘药材质量差异成分。方法 采用菲罗门Titank C₁₈色谱柱（150 mm×2.1 mm，1.8 μm），以体积分数乙腈（A）-0.2%甲酸水溶液（B）为流动相，梯度洗脱（0～5 min，5%～12% A；5～8 min，12%～20% A；8～15 min，20% A；15～18 min，20%～29% A；18～20 min，29%～48% A；20～25 min，48%～70% A；25～28 min，70%～85% A；28～30 min，85%～5% A），体积流量为0.3 mL/min，柱温为30 ℃，进样量为0.4 μL，采用定时波长检测（0～13.2 min，265 nm；13.2～20 min，260 nm；20～21.5 min，270 nm；21.5～22 min，275 nm；22～30 min，280 nm）。采用中药指纹图谱相似度评价软件进行相似度评价，结合化学计量学分析，同时测定连翘酯苷A、芦丁、连翘苷、松脂醇、连翘脂素的含量。结果 建立了连翘药材的指纹图谱，以6号峰为参照峰，共标记了11个共有峰，用对照品比对法指认出5个色谱峰，分别为峰6（连翘酯苷A）、峰7（芦丁）、峰9（连翘苷）、峰10（松脂醇）、峰11（连翘脂素）。24批连翘药材样品指纹图谱相似度均在0.99以上。聚类分析将24批连翘样品分为3类，不同产地来源各自聚为一类。主成分分析表明，不同产地来源的连翘样品间存在差异。连翘酯苷A、芦丁、连翘苷、松脂醇、连翘脂素的质量分数分别为59.286 5～114.849 2、2.653 6～4.055 6、7.133 3～11.911 1、0.300 7～1.614 6、0.089 3～0.838 2 μg/g，经方法学考察，各成分呈现良好的线性关系，平均加样回收率在104.30%～111.48%，RSD在1.86%～2.96%。通过OPLS-DA项下的变量权重值（variable importance projection，VIP）分析筛选出连翘酯苷A等3个成分可作为不同产地连翘的差异标志物。结论 建立的UPLC指纹图谱和多成分含量测定方法稳定、可靠，可为连翘药材的质量评价与控制提供科学依据及参考。

Objective To establish an ultra-high performance liquid chromatography (UPLC) fingerprint and multi-component content determination method for Forsythiae Fructus from different sources, and to evaluate the quality of Forsythiae Fructus from different sources in combination with chemometrics methods, so as to provide technical method reference and basic data for its quality control research. Methods Phenomenex Titank C₁₈ chromatographic column (150 mm × 2.1 mm, 1.8 μm ) was used, with acetonitrile (A) - 0.2% formic acid water (B) solution as mobile phase, gradient elution (0—5 min, 5%—12% A; 5～8 min, 12%—20% A; 8—15 min, 20% A; 15—18 min, 20%—29% A; 18—20 min, 29%—48% A; 20—25 min, 48%—70% A; 25—28 min, 70%—85% A; 28—30 min, 85%—5% A). The flow rate was 0.3 mL/min, the column temperature was 30 °C, the injection volume was 0.4 μL, and time-programmed wavelength detection wavelength was set at a fixed wavelength (0—13.2 min, 265 nm; 13.2—20 min, 260 nm; 20—21.5 min, 270 nm; 21.5—22 min, 275 nm; 22—30 min, 280 nm). The fingerprints of Forsythiae Fructus from different sources were constructed, and the contents of forsythiaside A, rutin, forsythin, pinoresinol and forsythigenin were determined by similarity analysis and chemometrics analysis. Results The fingerprint of Forsythia suspensa was established, and taking peak 6 as the reference peak, 11 common peaks were marked. A total of five chromatographic peaks were identified by reference substance comparison method, which were peak 6 (forsythiaside A), peak 7 (rutin), peak 9 (forsythin), peak 10 (pinoresinol) and peak 11 (forsythigenin). The similarity of fingerprints of 24 batches of Forsythiae Fructus samples was above 0.99. Cluster analysis divided 24 batches of Forsythiae Fructus samples into three categories, and different origins were clustered into one category. Principal component analysis showed that there were differences among Forsythiae Fructus samples from different origins. The mass fraction ranges of forsythiaside A, rutin, forsythin, pinoresinol and forsythigenin were 59.286 5—114.849 2、2.653 6—4.055 6、7.133 3—11.911 1、0.300 7—1.614 6、0.089 3—0.838 2 μg/g, respectively. The methodological investigation showed that each component showed a good linear relationship. The average recovery was between 104.30 % and 111.48 %, and the RSD was between 1.86 % and 2.96 %. Three components such as forsythiaside A were screened by VIP analysis under OPLS-DA, which could be used as differential markers of Forsythiae Fructus in different producing areas. Conclusion The UPLC fingerprint and multi-component content determination method established in this study are stable and reliable, which can provide scientific basis and reference for the quality evaluation and control of Forsythiae Fructus.

R286.2

河北省省级科技计划项目资助（21372503D）