目的 建立真武汤HPLC-ELSD指纹图谱，采用化学模式识别法对其进行分析和筛选出真武汤的标志性成分，并以其为指标建立真武汤含量测定方法。方法 采用HPLC-ELSD法建立16批真武汤指纹图谱，运用中药色谱指纹图谱相似度评价系统（2012版）进行相似度评价，确定共有峰及其归属，运用聚类分析（CA）、主成分分析（PCA）、正交偏最小二乘法判别分析（OPLS-DA），筛选出真武汤的指标性成分。结果 建立了真武汤指纹图谱，确认了38个共有峰，相似度均>0.95；CA、PCA、OPLS-DA结果一致，将样品分为3类；不同批次真武汤样品差异贡献较大的指标成分指认了11个，即苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、猪苓酸C、茯苓酸、白术内酯Ⅱ、白术内酯Ⅲ、羟基芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷；6-姜酚、6-姜烯酚为生姜主要活性成分，故将以上13个成分作为真武汤指标性成分，各色谱峰分离度较好，线性关系良好，平均加样回收率为96.46%～99.80%，RSD≤3.15%；16批次样品中苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、猪苓酸C、茯苓酸、白术内酯Ⅱ、白术内酯Ⅲ、羟基芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷、6-姜酚、6-姜烯酚的质量分数分别为283.93～576.86、25.05～147.39、62.96～303.37、31.24～131.27、9.76～44.04、32.15～83.55、76.55～333.13、17.48～146.61、456.58～1554.14、3 322.48～5 590.01、158.21～556.50、525.85～582.92、68.52～74.73 mg/g。结论 指纹图谱与PCA、CA、OPLS-DA相结合可全面地评价真武汤质量，该方法稳定、可靠，为真武汤质量评价提供参考。

Objective To establish HPLC-ELSD fingerprint of Zhenwu Decoction(ZWD), screen out the signature components of ZWD through chemical pattern recognition, so as to establish the content determination method of ZWD based on this index. Methods The fingerprint of 16 batches of ZWD was established by HPLC-ELSD method. The similarity evaluation system of traditional Chinese medicine chromatographic fingerprint (2012 Version) was used for similarity evaluation to determine the common peaks and its attribution. Cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to select the index components of ZWD. Results The fingerprint of ZWD was established, 38 common peaks were confirmed, and the similarity was > 0.95. The results of CA, PCA and OPLS-DA were consistent and the samples were divided into three categories. Benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, polyporenic acid C, pachymic acid, atractylenolide Ⅱ, atractylenolide Ⅲ, oxypaeoniflorin, albiflorin, paeoniflorin and benzoylpaeoniflorin were identified as the 11 index components with significant difference contribution in different batches of ZWD samples. 6-Gingerol and 6-shogaol were the main active components of ginger, so the above 13 components were taken as the index components of ZWD. The chromatographic peak separation degree and linear relationship were good. The average recovery rate was 96.46%-99.80%, RSD ≤ 3.15%. The mass fraction range of benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, polyporenic acid C, pachymic acid, atractylenolide Ⅱ, atractylenolide Ⅲ, oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin, 6-gingerol, 6-shogaol in 16 batches were 283.93-576.86, 25.05-147.39, 62.96-303.37, 31.24-131.27, 9.76-44.04, 32.15-83.55, 76.55-333.13, 17.48-146.61, 456.58-1554.14, 3 322.48-5 590.01, 158.21-556.50, 525.85-582.92 and 68.52-74.73 mg/g, respectively. Conclusion The fingerprint combined with PCA, CA and OPLS-DA can comprehensively evaluate the quality of ZWD. This method is stable and reliable, providing reference for the quality evaluation.

R286.02

国家自然科学基金资助项目（81473368）；河南省中医药科学研究专项课题（2016ZY2020）；河南省中医药科学研究专项课题（2017 ZY2036）