目的 建立指纹图谱和多指标定量与化学计量学相结合的湿热痹片质量评价方法。方法 采用Waters Symmetry C₁₈色谱柱（250 mm×4.6 mm，5 μm），柱温30℃；检测波长分别为303 nm（检测桑皮苷A、桑皮苷F、桑辛素M）和270 nm（检测连翘酯苷B、连翘酯苷A、连翘苷、苍术素醇、白术内酯II、苍术素）；流动相为乙腈-0.2%磷酸水溶液，梯度洗脱，体积流量1.0 mL/min。利用中药色谱指纹图谱相似度评价系统（2012.130723版）建立湿热痹片的HPLC指纹图谱，确定共有峰并进行相似度评价；并对桑皮苷A、桑皮苷F、桑辛素M、连翘酯苷B、连翘酯苷A、连翘苷、苍术素醇、白术内酯II、苍术素的含量测定方法进行方法学验证；基于指纹图谱共有峰峰面积测定结果，采用聚类分析和主成分分析等化学计量学方法对不同批次的湿热痹片进行质量评价。结果 湿热痹片HPLC指纹图谱确认了16个共有峰，指认9个共有峰，10批湿热痹片样品相似度均大于0.95，相似度良好；9种成分在各自的质量浓度范围内线性关系良好（r² ≥ 0.999 1），平均加样回收率分别为98.87%、97.44%、97.94%、98.39%、100.13%、99.06%、96.80%、98.44%、99.15%，RSD分别为1.42%、1.17%、1.30%、0.91%、0.86%、1.23%、1.08%、1.37%、0.79%；10批次样品中桑皮苷A、桑皮苷F、桑辛素M、连翘酯苷B、连翘酯苷A、连翘苷、苍术素醇、白术内酯II、苍术素的质量浓度分别在0.192~0.289、0.057~0.095、0.113~0.158、0.309~0.375、1.537~1.916、0.478~0.596、0.049~0.072、0.279~0.354、0.629~0.759 mg/g。10批湿热痹片聚为2类；主成分1~6是影响湿热痹片质量评价的主要因子。结论 所建立的方法操作便捷、结果准确、重复性好，可用于湿热痹片的质量控制和评价。

Objective To establish a quality evaluation method of Shirebi Tablet based on HPLC fingerprints, quantitative analysis of multi-components and chemometrics analysis. Methods The chromatographic column was Waters Symmetry C₁₈ column (250 mm×4.6 mm, 5 μm), and the column temperature was set at 30℃. The detection wavelength was set at 303 nm (for mulberroside A, mulberroside F and moracin M) and 270 nm (for forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide II, and atractylodin). The mobile phase was composed of acetonitrile-0.2% phosphate acid solution in gradient elution manner at a flow rate of 1.0 mL/min. The HPLC fingerprint of Shirebi Tablet was established, the common peaks were determined by similarity evaluation system for chromatographic fingerprint of TCM (Version 2012.130723), and the similarity was calculated. The content determination methods for mulberroside A, mulberroside F, moracin M, forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide II, and atractylodin were validated. The chemometrics methods such as cluster analysis and principal component analysis were used to evaluate the quality of Shirebi Tablet from different batches based on the results of fingerprint common peak area. Results The fingerprint of Shirebi Tablet was established. Sixteen common peaks were identified. The similarity of fingerprints of 10 batches of Shirebi Tablet was more than 0.95. Nine components had good linear relationship in the range of mass concentration (r² ≥ 0.999 1), and the average recoveries were 98.87%, 97.44%, 97.94%, 98.39%, 100.13%, 99.06%, 96.80%, 98.44% and 99.15% with the RSDs of 1.42%, 1.17%, 1.30%, 0.91%, 0.86%, 1.23%, 1.08%, 1.37% and 0.79%, respectively. The concentrations of mulberroside A, mulberroside F, moracin M, forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide II, and atractylodin in 10 batches were 0.192-0.289, 0.057-0.095, 0.113-0.158, 0.309-0.375, 1.537-1.916, 0.478-0.596, 0.049-0.072, 0.279-0.354, and 0.629-0.759 mg/g, respectively. The results of cluster analysis showed that 10 batches of Shirebi Tablet were clustered into two groups, and the results of principal component analysis showed that the principal components 1-6 were the main factor affecting the quality evaluation of Shirebi Tablets. Conclusion The method is simple, accurate and reproducible, which can be used for the quality control and evaluation of Shirebi Tablet.

R286.02

国家自然科学基金面上项目（81872236）；天津市自然科学基金项目（18JCYBJC95100）