女儿茶HPLC指纹图谱及清除自由基活性谱效关系

Correlation Analysis Between HPLC Fingerprint and Free Radical-scavenging Activity of Rhamnus heterophylla Leaves

目的：研究女儿茶Rhamnus heterophylla HPLC指纹图谱与其清除自由基活性的关系。方法：采用高效液相色谱法建立8批不同产地、采收期女儿茶指纹图谱,在传统相似度评价基础上,采用Origin Pro 2017软件进行层次聚类分析（hierarchical cluster analysis,HCA）和主成分分析（principal component analysis,PCA）对其指纹图谱中的共有峰进行评价;1,1-二苯基-2-三硝基苯肼（DPPH）法研究其清除自由基活性;偏最小二乘回归分析（partial least squares regression,PLSR）和灰色关联度分析（grey relational analysis,GRA）研究谱效关系。结果：建立了8批女儿茶HPLC指纹图谱,确定了11个共有峰,相似度在0.948-0.976。采用对照品比对方法指认了其中6个峰：x1为原儿茶酸,x2为木犀草苷,x5为槲皮素,x8为山柰酚,x10为大黄素8-O-α-L-鼠李糖苷,x11为大黄素。样本可聚为4类。8批女儿茶均有不同程度清除自由基活性。灰色关联度分析结果显示共有峰与清除自由基活性的关联度大小为x11〉x10〉x8〉x1〉x9〉x4〉x3〉x2〉x6〉x5〉x7。偏最小二乘回归分析建立的回归方程为Y=99.769 17-0.357 49x1-0.001 36x2-0.002 65x3＋0.059 63x4＋0.011 81x6＋0.010 63x7-0.006 99x8-0.009 14x9＋0.054 27x10＋0.022 75x11。结论：女儿茶清除自由基活性是多组分联合效应的结果。

Objective： To explore the correlation between high performance liquid chromatography（ HPLC） fingerprint and free radical-scavenging activity of Rhamnus heterophylla leaves. Method： Fingerprint of eight batches of R. heterophylla leaves from different regions and harvest times was established by HPLC. Origin Pro 2017 software was used for hierarchical cluster analysis（ HCA） and principal component analysis（ PCA） to evaluate the common peaks in the fingerprint on the basis of traditional similarity evaluation method. The free radical-scavenging activity was determined by 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-（ 2,4,6-trinitrophenyl） hydrazyl（ DPPH） assay. Partial least squares regression（ PLSR） and grey relational analysis（ GRA） were used to study the relationship between HPLC fingerprint and free radical-scavenging activity. Result：The HPLC fingerprint of eight batches of R. heterophylla leaves was established, and 11 common peaks were determined,with the similarity of 0. 948-0. 976. Six peaks among common peaks were identified by comparing to reference substances： x1, x2, x5, x8, x（10）, x（11） referred to protocatechic acid, luteoloside, quercetin,kaempferol,emodin-8-O-α-L-rhamnoside, and emodin, respectively. The samples were clustered into four categories. All of the samples had different degrees of scavenging free radical activity. The result of GRA showed that the order of correlation was x11 x10 x8 x1 x9 x4 x3 x2 x6 x5 x7. The regression equation established by PLSR analysis was Y = 99. 769 17-0. 357 49 x1-0. 001 36 x2-0. 002 65 x3＋ 0. 059 63 x4＋0. 011 81 x6＋ 0. 010 63 x7-0. 006 99 x8-0. 009 14 x9＋ 0. 054 27 x10＋ 0. 022 75 x11. Conclusion： The free radicalscavenging activity of R. heterophylla leaves was the result of the combined effects of multiple components.