[关键词]
[摘要]
目的 建立生菟丝子Cuscutae Semen与酒菟丝子UPLC指纹图谱,并结合多元统计分析和定量测定研究菟丝子酒炙前后化学成分的变化,为菟丝子质量评价提供参考。方法 采用UPLC法进行测定,色谱柱为Thermo AccucoreTM C18柱(100 mm×4.6 mm,2.6 μm),流动相为乙腈-0.1%磷酸水溶液,柱温30℃,梯度洗脱,进样体积为2 μL,体积流量为0.4 mL/min,采用分段波长,建立15批酒菟丝子及15批生菟丝子指纹图谱。通过相似度评价、层次聚类分析(hierarchical clustering analysis,HCA)、主成分分析(principal component analysis,PCA)来对指纹图谱进行分析评价,同时对9个主要成分进行定量测定,寻找炮制前后差异成分。结果 建立的UPLC指纹图谱中,共匹配了19个共有峰,宁夏产区与内蒙产区的生菟丝子和酒菟丝子相似度均较高,HCA中可将酒菟丝子与生菟丝子明显区分,PCA提取出4个主成分,能够明显区分菟丝子生品及炮制品。正交偏最小二乘-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)亦可将二者明显区分,并筛选出菟丝子炮制前后的8个差异性成分,根据VIP值排序为峰1、10、2、5、8(隐绿原酸)、4(新绿原酸)、18(槲皮素)、17(异绿原酸C)。含量测定结果表明,酒炙后菟丝子中金丝桃苷、异槲皮苷、槲皮素、新绿原酸、隐绿原酸含量增加,紫云英苷、山柰酚、绿原酸、异绿原酸C含量降低。结论 建立的UPLC指纹图谱和含量测定结果对菟丝子及其炮制品的质量控制及整体性评价具有重要意义。
[Key word]
[Abstract]
Objective To establish UPLC fingerprints of Cuscutae Semen and Cuscutae Semen stir-frying with wine, and to study the changes of chemical constituents of Cuscutae Semen before and after processing by multivariate statistical analysis and quantitative determination, so as to provide reference for quality evaluation of Cuscutae Semen. Methods The fingerprints of 15 batches of Cuscutae Semen stir-frying with wine and 15 batches of raw Cuscutae Semen were established and determined by UPLC. The chromatographic column was Thermo AccucoreTM C18 (100 mm × 4.6 mm, 2.6 μm), the mobile phase was acetonitrile-0.1% phosphoric acid water, column temperature was 30℃, gradient elution, the injection volume was 2 μL, and volume flow rate was 0.4 mL/min. Segmented wavelengths were used. The fingerprint was analyzed and evaluated by similarity evaluation, hierarchical cluster analysis (HCA), principal component analysis (PCA), and quantitative determination of nine main components to find the difference components before and after processing. Results In the established fingerprint of Cuscutae Semen, a total of 19 common peaks were matched. The products from Ningxia and Inner Mongolia showed a high similarity. In the cluster analysis, Cuscutae Semen could be clearly distinguished from Cuscutae Semen. The four principal components extracted by PCA can be used to distinguish raw and processed products. Orthogonal partial least squares-discriminant analysis (OPLS-DA) can also be used to distinguish the eight different components of Cuscutae Semen before and after processing, according to VIP values, they were ranked as peaks 1, 10, 2, 5, 8 (cryptochlorogenic acid), 4 (neochlorogenic acid), 18 (quercetin), and 17 (isochlorogenic acid C). The results showed that the contents of hyperin, isoquercitrin, quercetin, neochlorogenic acid and cryptochlorogenic acid increased, while the contents of shikonin, kaempferol, chlorogenic acid and isochlorogenic acid C decreased. Conclusion The established UPLC fingerprint and content determination results are of great significance for the quality control and overall evaluation of Cuscutae Semen and its processed products.
[中图分类号]
R283.1
[基金项目]
国家重点研发计划(2017YFC1701500);国家重点研发计划(2017YFC1701502);国家重点研发计划(2017YFC1701504);山东省中药材及中药饮片标准研究;中央转移支付项目“道地药材生态种植及质量保障项目”;济南市农业应用技术创新计划(CX202112);山东省科技型中小企业创新能力提升工程“道地黄芩生态种植技术及产业化研究”(2022TSGC1059);中央本级重大增减支项目(2060303)