[关键词]
[摘要]
目的 建立曲花紫堇Corydalis curviflora的HPLC指纹图谱,并通过所建立的指纹图谱方法对紫堇属其他14种药材进行对比研究。方法 HPLC-DAD指纹图谱条件:Agilent ZORBAX Extend C18(250 mm×4.6 mm,5 μm);流动相为甲醇-0.2%磷酸水溶液,梯度洗脱;体积流量1.0 mL/min;检测波长289 nm;柱温30 ℃。结合聚类分析(combine cluster analysis,HCA)、主成分分析(principal component analysis,PCA)及正交偏最小二乘法-判别分析(orthogonal partial least square discriminant analysis,OPLS-DA)区分与其极易混淆的暗绿紫堇。结果 首次建立了曲花紫堇HPLC指纹图谱,相似度为0.853~0.996,标定18个共有峰,通过对照品比对指认出新绿原酸、绿原酸、原阿片碱、紫堇灵、乙酰紫堇灵、芦丁、紫云英苷、烟花苷、槲皮素、山柰酚10个共有峰;14种同属药材与曲花紫堇对照指纹图谱相似度在0.078~0.686。通过化学模式识别可区分曲花紫堇和暗绿紫堇,二者有6个差异性标志物。结论 建立的指纹图谱方法合理、准确,结合化学计量学方法,能有效地鉴别曲花紫堇及同属药材,寻找不同紫堇质量差异成分,为紫堇属不同药材的质量控制和品质评价提供科学依据和参考。
[Key word]
[Abstract]
Objective To establish the HPLC fingerprint of Corydalis curviflora and analyze the other 14 medicinal materials of Corydalis by the established fingerprint method. Methods The HPLC analysis was performed on a Agilent ZORBAX ExtendC18 (250 mm × 4.6 mm, 5 μm) , the mobile phase was methanol-0.2% phosphoric acid aqueous solution for gradient elution, the volume flow rate was 1.0 mL/min, the detection wavelength was 289 nm, and the column temperature was 30 ℃. The combine cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA) were used to distinguish C. melanochlora which is easy to be confused with C. curviflora. Results The HPLC fingerprint of C. curviflora was established for the first time, the similarity was 0.853—0.996, and 18 common peaks were calibrated. Through comparison with the reference substance, 10 components were identified: neochlorogenic acid, chlorogenic acid, protopine, corynoline, acetyl corynoline, rutin, astragalin, nicotiflorin, quercetin, kaempferol; The similarity of the fingerprints of 14 kinds of medicinal materials and C. curviflora was in the range of 0.078—0.686. The chemical pattern recognition can distinguish C. curviflora and C. melanochlora, with six differential markers between them. Conclusion The established fingerprint method is reasonable and accurate, and can effectively identify C. curviflora and the same genus of medicinal materials combined with chemometric methods to find the differential components of different Corydalis medicinal materials, which provide scientific basis and reference for the quality control and quality evaluation of different Corydalis medicinal materials.
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[基金项目]
国家自然科学基金项目(82173928);国家重点研发计划课题项目(2019YFC1712302);国家重点研发计划课题项目(2019YFC1712305);四川省科技厅应用基础项目(2020YJ0369)