目的 建立窝儿七HPLC指纹图谱，并同时测定19个批次不同产地窝儿七中苦鬼臼毒素葡萄糖二苷、苦鬼臼毒素葡萄糖苷、4'-去甲基鬼臼毒素、鬼臼毒素葡萄糖苷、山柰酚葡萄糖苷、鬼臼毒素、arabeline、鬼臼毒酮、山荷叶素葡萄糖苷、槲皮素、山柰酚和山荷叶素12种化学成分含量，为窝儿七质量控制提供科学依据。方法 采用COSMOSIL-C₁₈色谱柱（250 mm×4.6 mm，5 μm），以甲醇（A）-0.4%磷酸水溶液（B）为流动相；梯度洗脱，体积流量1.0 mL/min，柱温30℃；检测波长300 nm，进样量10 μL；建立窝儿七药材指纹图谱，测定12个成分含量，对结果进行聚类分析。结果 19批窝儿七HPLC指纹图谱有16个共有峰，各相似度均在0.9以上，12个化学成分在各自范围内线性关系良好（r² ≥ 0.999 0），精密度、重复性RSD<2%，在24 h内稳定性较好（RSD<2%），加样回收率均在99.27%~100.30%，RSD均在1.03%~1.98%。19批样品中上述12个成分的含量测定结果及聚类分析结果可知不同产地窝儿七质量有所差异。结论 该方法简单、灵敏、准确，为窝儿七的鉴别及质量评价提供了全面的参考依据。

Objective To establish the HPLC fingerprints of Diphylleia sinensis from different habitats and determine the content of 12 chemical ingredients of picropodophillotoxin-4-O-β-D-glucopyranosy-(1→6)-β-D-glucopyranoside, picropod ophyllotoxin 4-O-glucoside, 4'-demethylpodophyllotoxin, podophyllotoxin-4-O-β-D-glucoside, kaempferol-3-O-β-D-glucoside, podophyllotoxin, arabeline, podophyllotoxone, diphyllin-4-O-β-D-glucoside, quercetin, kaempferol, and diphyllin for providing a scientific basis for the quality control of D. sinensis. Methods COSMOSIL-C₁₈ column (250 mm×4.6 mm, 5 μm) was used for gradients elution with MeOH (A) −0.4% phosphoric acid solution (B) as mobile phase. Working conditions were as follows:the column temperature was 30℃, the flow rate was 1 mL/min, the detection wavelength was 300 nm, and the injection volume was 10 μL. HPLC fingerprint of D. sinensis was established and 12 components were determined. The results were analyzed by cluster analysis. Results The HPLC fingerprint with 16 common peaks of D. sinensis was established, and the similarities of samples were over 0.9. The linear relationship of 12 components was good (r² ≥ 0.999 0), RSD of precision and repeatability was less than 2%, and the stability was also good with in 24 h (RSD<2%). The average recoveries (n=6) of 12 components were between 99.27% and 100.3%, and the RSD were in the range of 1.03%-1.98%. The results of the content determination and cluster analysis of twelve components showed that D. sinensis in different habitats were different from each other. Conclusion This method was simple, sensitive and accurate, which provided a comprehensive reference for the identification and quality evaluation of D. sinensis.

R286.12

陕西省重点研发计划项目（2019ZDLSF04-03-02）；陕西中医药大学学科创新团队项目（2019-YL12）