藏药圆柏HPLC指纹图谱的建立及不同品种模式识别研究

目的 建立圆柏药材HPLC化学指纹图谱，并比较不同基原品种的差异。方法 采用HPLC法，色谱柱为WondaSil^?C₁₈-WR(250 mm×4.6 mm,5μm)，流动相为甲醇-0.2%磷酸水溶液，梯度洗脱，体积流量为1.0 mL/min，检测波长为270 nm。对49批圆柏药材进行指纹图谱构建，采用“中药色谱指纹图谱相似度评价系统”软件(2012年130723版)进行相似度评价，运用主成分分析(principalcomponentanalysis,PCA)、偏最小二乘法判别分析(partialleastsquares discriminant analysis,PLS-DA)和方差分析研究圆柏5个主流品种的化学成分差异。结果 建立了圆柏HPLC指纹图谱，标定15个共有色谱峰，相似度0.648～0.975，仅有26批相似度大于0.9。鉴定4个峰为槲皮苷、穗花衫双黄酮、罗汉松双黄酮A和扁柏双黄酮。圆柏5个品种之间的化学成分存在差异，并发现7个种间显著差异成分。结论 建立的化学指纹图谱稳定可靠，可为其质量控制及评价提供参考。

Establishment of HPLC fingerprint of Tibetan medicine Juniperri Caulis et Folium and pattern recognition of its different species

Objective To establish HPLC fingerprints of Yuanbai (Juniperri Caulis et Folium) and compare the differences between different original species. Methods HPLC was performed on WondaSil^® C₁₈-WR (250 mm×4.6 mm, 5 µm). The mobile phase was methanol-0.2% phosphoric acid aqueous solution with the detection wavelength of 270 nm and volume flow rate of 1.0 mL/min for gradient elution. Similarity of fingerprints of 49 batches samples were evaluated by chromatographic fingerprint similarity evaluation software. The chemical composition differences of five species were studied by principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and One-Way ANOVA. Results The HPLC fingerprint of Juniperri Caulis et Folium was established and 15 common peaks were calibrated. Among them, the similarity of fingerprints ranged from 0.648 to 0.975, only 26 batches were above 0.9. Four components were identified as quercetin, amentoflavone, podocarpusflavone A and hinokiflavone. There were differences in chemical components among five species, and seven inter-species differential components were found. Conclusion The established fingerprint is stable and reliable, which can provide the basis for its comprehensive evaluation and quality control.