目的 对不同产地初加工方式处理杜仲叶进行HPLC指纹图谱的比较研究，阐明不同初加工方式对杜仲叶化学成分的影响，以期筛选出适合杜仲叶的产地初加工方式。方法 对所采集的杜仲鲜叶以不同的产地初加工方式进行加工处理，并采用HPLC法，建立用以评价初加工方式优劣的指纹图谱体系。色谱条件：以Promosil C¹⁸（250 mm×4.6 mm，5 μm）为色谱柱，乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱，扫描波长200～400 nm，检测波长208 nm，柱温25 ℃，体积流量0.8 mL/min。结果 系统研究了杜仲叶的不同产地初加工方式，并建立了评价其优劣的杜仲叶HPLC指纹图谱，确定了22个主要共有峰，但是各共有峰峰面积相差较大，绿原酸、京尼平苷酸等主要有效成分的量有明显差异。结论 建立的HPLC指纹图谱分析法可同时用于有效成分的定量测定，该方法重复性好、可操作性强，可较全面地反映不同产地初加工方式处理的杜仲叶间的差异。

Objective To study the optimal extraction for Hirudo by wet ultrafine grinding technology. Methods On the basis of single-factor experiments, the response surface analysis was made for exploring the optimal extraction conditions by wet ultrafine grinding technology for Hirudo, with solid-liquid ratio, soaking time, extraction temperature, and grinding time as independent variable and the anticoagulant activity as the response value. According to Box-Behnken principles, the response surface analysis method with four factors and three levels was used. Results The optimum extraction condition for Hirudo by wet ultrafine grinding technology was as follows: solid-liquid ratio 1∶12, soaking time 4.9 h, extraction temperature 6.5 ℃, grinding time 12 min; The anticoagulant activity could reach 20.13 U/g. Conclusion The extraction by wet ultrafine grinding technology for Chinese materia medica Hirudo is stable and feasible, and it could be further promoted.

国家“重大新药创制”科技重大专项（子课题2011ZX09401-308-008）；国家科技重大专项子课题：中药大品种脑心通胶囊技术改造研究（2011ZX09201-201-26）；陕西省科技统筹创新工程计划课题（2011KTZB03-01-04）