[关键词]
[摘要]
目的 建立金骨莲胶囊HPLC指纹图谱,并结合化学模式识别方法对其进行质量一致性评价,为该制剂的质量控制提供参考。方法 采用ACE Excel 5 C18-PFP(250 mm×4.6 mm,5 μm)色谱柱进行检测;流动相为乙腈-0.1%磷酸水溶液,进行梯度洗脱;体积流量为0.8 mL/min;柱温为40 ℃;检测波长为210 nm;进样量为10 μL;建立12批金骨莲胶囊指纹图谱。对12批金骨莲胶囊指纹图谱进行相似度评价、层次聚类分析(hierarchical clustering analysis,HCA)和主成分分析(PCA),并结合正交偏最小二乘-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)寻找不同批次金骨莲胶囊的差异成分。结果 建立的金骨莲胶囊HPLC指纹图谱共标定了19个共有峰,其中2、8、11号峰归属于汉桃叶,5、7、10、12、15号峰归属于大血藤,16、17、19号峰归属于透骨香,1号峰为5味药材共有,3号峰归属于八角枫和大血藤,13号峰归属于大血藤和透骨香,4、9、14、18号峰归属于大血藤和汉桃叶,6号峰归属于大血藤、汉桃叶和透骨香。通过与对照品对比指认出2、3、6、7、9、12、13、15、16号峰分别为富马酸、没食子酸、原儿茶酸、红景天苷、绿原酸、香草酸、表儿茶素、鹅掌楸苷、滇白珠苷A。12批金骨莲胶囊指纹图谱相似度均在0.910以上,HCA、PCA 2种分析方法均把样品分为了3类,结合OPLS-DA筛选出了导致各批次之间产生差异的7个差异性标志物,分别为15(鹅掌楸苷)、14、6(原儿茶酸)18、16(滇白珠苷A)、19、9(绿原酸)号色谱峰。结论 该分析方法简单可行、具有良好精密度、重复性和稳定性,建立的指纹图谱可为金骨莲胶囊的质量评价提供参考。
[Key word]
[Abstract]
Objective To establish the HPLC fingerprint of Jingulian Capsules (JGLC), and evaluate its quality consistency by combining with the chemical pattern recognition, in order to provide reference for its quality control. Methods The detection was performed on ACE Excel 5 C18-PFP (250 mm×4.6 mm, 5 μm) column with the mobile phase consisting of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution at a flow rate of 0.8 mL/min, the column temperature was 40 ℃, the detection wavelength was 210 nm, and the injection volume was 10 μL to establish the HPLC fingerprints of 12 batches of JGLC. Then the further assessment of 12 batches of samples were carried out by similarity evaluation, hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to find the different components of different batches of JGLC. Results The HPLC fingerprints of JGLC was set up with 19 common peaks, peaks 2, 8 and 11 belonged to Hantaoye [Schefflerae Leucanthae Ramulus et Folium (SLRF)], peaks 5, 7, 10, 12, and 15 belonged to Daxueteng [Sargentodoxae Caulis (SC)], peaks 16, 17 and 19 belonged to Touguxiang [Gaultheriae Yunnanensis Herba seu Radix (GYHR)], peak 1 belonged to all of the five herbs, peak 3 belonged to Bajiaofeng (Alangii Radix) and SC, and peak 13 belonged to SC and GYHR, peaks 4, 9, 14 and 18 belonged to SC and GYHR, peak 6 belonged to SC, SLRF and GYHR, and nine common peaks of them were identified, namely, fumaric acid, gallic acid, protocatechuic acid, salidroside, chlorogenic acid, vanillic acid, epicatechin, liriodendrin and gaultheroside A (corresponding to peaks 2, 3, 6, 7, 9, 12, 13, 15 and 16) by comparing with the reference substances. The similarities of 12 batches of JGLC were all above 0.910. The samples were classified into three clusters by HCA and PCA, and seven differential markers that caused differences in the 12 batches samples were found by OPLS-DA. Conclusion The proposed fingerprint method is simple, accurate, reproducible and stable, which could provide scientific reference for the quality evaluation of JGLC.
[中图分类号]
R283.6
[基金项目]
国家重点研发计划项目(2018YFC1708100);国家自然科学基金项目(81960763);贵州省科技计划项目(黔科合平台人才[2019]5407\5660);2020年国家级大学生创新创业训练计划项目(10660-462);贵阳市科研创新团队项目(筑科合同[2017]30-29号)