马甲子总三萜的HPLC指纹图谱建立、聚类分析和主成分分析及含量测定  

Establishment of HPLC Fingerprint,Cluster Analysis,Principal Component Analysis and Content Determination of Paliurus ramosissimus Total Triterpenes

作　　者：陈旺 詹雁[2] 王美慧 谭镭[2] 徐超群[2] CHEN Wang;ZHAN Yan;WANG Meihui;TAN Lei;XU Chaoqun(College of Pharmacy,Chengdu University of TCM,Chengdu 611137,China;Sichuan Provincial Academy of Chinese Medicine Science,Chengdu 610041,China)

机构地区：[1]成都中医药大学药学院,成都611137 [2]四川省中医药科学院,成都610041

出　　处：《中国药房》2021年第2期201-206,共6页China Pharmacy

基　　金：国家科技重大专项(民口)课题(No.2018ZX09731-013);四川省重大科技专项课题(No.2017SZDZX0005);四川省省级科研院所基本科研业务费项目。

摘　　要：目的:建立马甲子总三萜的指纹图谱,进行聚类分析和主成分分析,并测定其中主要成分马甲子素的含量。方法:采用高效液相色谱法(HPLC)。色谱柱为Agilent PheHex,流动相为甲醇-0.05%磷酸水溶液(梯度洗脱),检测波长为320 nm,流速为1mL/min,柱温为30℃,进样量为5μL。以马甲子素为参照,绘制10批马甲子总三萜的HPLC指纹图谱,采用《中药色谱指纹图谱相似度评价系统(2012版)》进行相似度评价,确定共有峰;采用SPSS 26.0软件进行聚类分析和主成分分析。采用同一HPLC法测定马甲子素的含量。结果:10批马甲子总三萜共有6个共有峰,相似度均大于0.990;指认马甲子素1个特征峰。聚类分析结果显示,10批马甲子总三萜样品可聚为4类,其中S1为一类、S2为一类、S3~S6为一类、S7~S10为一类。主成分分析结果显示,前2个主成分的累积方差贡献率为99.430%。综合评分由高到低依次为S1>S9>S8>S7>S10>S2>S3>S5>S6>S4。马甲子素检测质量浓度的线性范围为33.7~844.0μg/mL(r=0.9999);精密度、重复性、稳定性(24 h)试验的RSD均小于2%,平均加样回收率为99.75%(RSD=1.13%,n=6);10批马甲子总三萜样品中马甲子素的平均含量为0.576%~0.712%。结论:所建HPLC指纹图谱和含量测定方法稳定、可靠,可用于马甲子的质量控制。OBJECTIVE:To establish fingerprint of Paliurus ramosissimus total triterpenes,and to conduct cluster analysis and principal component analysis,and to determine the content of the main component paliurusene.METHODS:HPLC method was adopted.The determination was performed on Agilent PheHex column with mobile phase consisted of methanol-0.05%phosphoric acid solution(gradient eluetion)at the flow rate of 1 mL/min.The detection wavelength was set at 320 nm,and column temperature was 30℃.The sample size was 5μL.Using paliurusene as reference,HPLC fingerprints of 10 batches of P.ramosissimus total triterpenes were drawn.Similarity evaluation was performed by using TCM Chromatographic Fingerprint Similarity Evaluation System(2012 edition),and the common peaks were confirmed.Cluster analysis and principle component analysis were performed by using SPSS 26.0 software.The content of paliurusene was determined by same HPLC method.RESULTS:There were totally 6 common peaks in HPLC fingerprint of 10 batches of P.ramosissimus total triterpenes.The similarity was more than 0.990;one of six common peaks was identified as paliurusene.The results of cluster analysis showed that10 batches of samples could be clustered into 4 categories,including S1,S2,S3-S6 and S7-S10.The results of principal component analysis showed that the accumulative variance contribution rate of primary 2 principal components was 99.430%.Comprehensive score ranking was S1>S9>S8>S7>S10>S2>S3>S5>S6>S4.The linear range of paliurusene concentration was 33.7-844.0μg/mL(r=0.9999).RSDs of precision,reproducibility and stability(24 h)tests were all lower than 2%.Average recovery was 99.75%(RSD=1.13%,n=6).The average contents of paliurusene in 10 batches of P.ramosissimus total triterpenes was 0.576%-0.712%.CONCLUSIONS:Established HPLC fingerprint and content determination method are reliable and stable,and can be used for the quality control of P.ramosissimus.

关 键 词：马甲子总三萜 马甲子素 高效液相色谱法 相似度评价 聚类分析 主成分分析 指纹图谱 含量测定 

分 类 号：R284[医药卫生—中药学]