目的 建立山慈菇Pleiones Pseudobulbus（冰球子）及其潜在代用品的HPLC指纹图谱，并结合化学模式识别寻找山慈菇（冰球子）及其潜在代用品的差异性成分，筛选山慈菇（冰球子）最佳代用品。方法 使用乙腈-0.1%磷酸水溶液作流动相进行梯度洗脱, Agilent ZORBAX SB-C₁₈色谱柱（250 mm×4.6 mm，5 μm），检测波长224 nm，进样量25 μL，柱温为30 ℃，体积流量为0.8 mL/min，建立40批样品HPLC指纹图谱，结合相似度评价、特征图谱、聚类热图分析、主成分分析（principalcomponent analysis，PCA）、正交偏最小二乘法判别分析（orthogonal partial least squares discriminant analysis，OPLS-DA）对特征成分进行数据分析。结果 40批样品共有5个共有峰，经对照品比对指认出了5个成分，分别为天麻素、对羟基苯甲醇、dactylorhin A、militarine、山药素III。聚类热图和PCA分析结果可将山慈菇（冰球子）及其潜在代用品进行区分，并结合相似度评价，初步判断带唇兰Tainia dunnii为山慈菇（冰球子）最佳代用品。不同基原样品的HPLC典型特征图谱结合OPLS-DA分析与PCA分析，筛选出山慈菇（冰球子）正品基原与其潜在代用品的主要差异性指标成分为loroglossin、dactylorhin A和militarine。结论 建立的指纹图谱结合化学模式识别方法可用于山慈菇（冰球子）及其潜在代用品的区分及山慈菇（冰球子）最佳代用品的筛选。

Objective To establish the HPLC fingerprint of Pleiones Pseudobulbus (Bing qiu zi) and potential substitutes, and combine with chemical pattern recognition to identify the differential components of Bing qiu zi and potential substitutes, so as to screen for the best substitutes of Bing qiu zi. Methods Gradient elution was performed using acetonitrile-0.1% phosphate acid water as the mobile phase, HPLC was used with Agilent ZORBAX SB-C₁₈column (250 mm×4.6 mm, 5 μm), the detection wavelength was 224 nm, the injection volume was 25 μL, the column temperature was 30 ℃, and the flow rate was 0.8 mL/min. HPLC fingerprint of 40 batches of samples was established, and the data of characteristic components was analyzed by combining similarity evaluation, HPLC specific chromatogram, cluster heatmap analysis, principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). Results There were a total of five common peaks in 40 batches of samples. After comparison with the reference substances, five components were identified, namely, gastrodin, p-hydroxybenzyl alcohol, dactylorhin A, militarine and batatasin III. The cluster heatmap analysis and PCA analysis results can distinguish between Bing qiu zi and potential substitutes, and combined with similarity evaluation, it was preliminarily determined that Tainia dunnii Rolfe was the best substitute for Bing qiu zi. OPLS-DA analysis combined with HPLC specific chromatogram and PCA analysis identified the differential components of Bing qiu zi and potential substitutes as loroglossin, dactylorhin A and militarine. Conclusion The fingerprint established in this study, combined with chemical pattern recognition methods, can be used to distinguish between Bing qiu zi and potential substitutes, as well as to screen for the best substitute of Bing qiu zi.

R286.12

国家自然科学基金资助项目（8227439）；国家中医药综合改革示范区科技共建项目（GZY-KJS-SD-2024-059）；山东省中药标准物质研究项目(2022-SDY0022); 郜文起全国老药工传承工作室建设项目(国中医药人教函 [2024] 255号)；全国名老中医药专家蒋立昶传承工作室建设项目（国中医药人教函 [2022] 75号）