藏药二十五味驴血丸的指纹图谱建立、含量测定及化学模式识别分析
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篇名: 藏药二十五味驴血丸的指纹图谱建立、含量测定及化学模式识别分析
TITLE: Fingerprint Establishment ,Content Determination and Chemical Pattern Recognition Analysis of Tibetan Medicine Ershiwuwei Luxue Pills
摘要: 目的:建立藏药二十五味驴血丸的指纹图谱,测定其中5种成分的含量,并进行化学模式识别分析。方法:采用高效液相色谱法(HPLC)。以秦皮乙素为参照,绘制10批藏药二十五味驴血丸的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;同时采用相同的HPLC法测定藏药二十五味驴血丸中5种成分的含量;采用SPSS19.0软件进行聚类分析和主成分分析。结果:10批藏药二十五味驴血丸共有11个共有峰,相似度均大于0.98;共指认了秦皮乙素、荭草苷、异牡荆素、异金雀花素和鞣花酸等5个共有峰。秦皮乙素、荭草苷、异牡荆素、异金雀花素、鞣花酸检测质量浓度的线性范围分别为1.232~11.092μg/mL(r=0.9996)、2.766~24.893μg/mL(r=0.9995)、1.400~12.600μg/mL(r=0.9998)、0.600~5.400μg/mL(r=0.9995)、49.447~445.025μg/mL(r=0.9994);精密度、稳定性(24h)、重复性试验的RSD均小于2%;平均加样回收率分别为101.29%(RSD=2.33%,n=3)、91.39%(RSD=1.22%,n=3)、90.28%(RSD=1.88%,n=3),98.76%(RSD=2.53%,n=3)、101.45%(RSD=2.84%,n=3)、100.44%(RSD=1.38%,n=3),100.91%(RSD=1.73%,n=3)、97.78%(RSD=2.07%,n=3)、99.15%(RSD=1.28%,n=3),100.27%(RSD=1.81%,n=3)、98.38%(RSD=1.89%,n=3)、101.92%(RSD=1.17%,n=3),95.50%(RSD=0.67%,n=3)、99.89%(RSD=0.38%,n=3)、100.10%(RSD=0.65%,n=3)。含量分别为0.175~0.310、0.351~0.632、0.274~0.395、0.186~0.278、6.956~8.636mg/g。聚类分析结果显示,10批藏药二十五味驴血丸样品可聚为两类,其中S1~S4为一类、S5~S10为一类。主成分分析结果显示,两个主成分的累积方差贡献率为89.178%。结论:所建指纹图谱稳定、可行,含量测定方法简便、准确、重复性好,结合化学模式识别分析可用于藏药二十五味驴血丸的质量控制。
ABSTRACT: OBJECTIVE:To establish fingerprint of Tibetan medicine Ershi wuwei luxue pills ,and determine the contents of 5 components,and to conduct chemical pattern recognition. METHODS :HPLC method was adopted. Using aesculetin as reference , HPLC fingerprint of 10 batches of Tibetan medicine Ershi wuwei luxue pills were drawn. The similarity evaluation was conducted by using Evaluation System of TCM Chromatogram Fingerprint Similarity (2012 edition),and the common peaks were determined. Same HPLC method was adopted to determine the contents of 5 components in Tibetan medicine Ershi wuwei luxue pills. The cluster analysis and principle component analysis were performed by using SPSS 19.0 software. RESULTS :Totally 11 common peaks were calibrated ,and the similarity was higher than 0.98. Five common peaks were identified ,as aesculetin ,orientin, isovitexin,isoscoparin and ellagic acid. The linear range of aesculetin ,orientin,isovitexin,isoscoparin and ellagic acid were 1.232-11.092 μg/mL(r=0.999 6),2.766-24.893 μg/mL(r=0.999 5),1.400-12.600 μg/mL(r=0.999 8),0.600-5.400 μg/mL(r= 0.999 5),49.447-445.025 μg/mL(r=0.999 4),respectively. RSDs of precision ,stability(24 h)and reproducibility tests were all lower than 2%. The average recoveries were 101.29%(RSD=2.33%,n=3),91.39%(RSD=1.22%,n=3),90.28%(RSD= 1.88%,n=3),98.76%(RSD=2.53%,n=3),101.45%(RSD=2.84%,n=3),100.44%(RSD=1.38%,n=3),100.91% (RSD=1.73%,n=3),97.78%(RSD=2.07%,n=3),99.15%(RSD=1.28%,n=3),100.27%(RSD=1.81%,n=3),98.38% (RSD=1.89% ,n=3),101.92%(RSD=1.17% ,n=3),95.50%(RSD=0.67% ,n=3),99.89%(RSD=0.38% ,n=3), 100.10%(RSD=0.65%,n=3),respectively. Their contents were 0.175-0.310, 0.351-0.632, 0.274-0.395, 0.186-0.278, 61932600 6.956-8.636 mg/g,respectively. Cluster analysis showed that 10 batches of Tibetan medicine Ershi wuwei luxue pills were clustered into two category ,with S1-S4 as one category and S5-S10 as one category. Principal component analysis showed that accumulative contribution rate of two principle components was 89.178%. CONCLUSIONS :Established fingerprint is stable and feasible ,and the method of content determination is simple , accurate and reproducible. They combined with chemical pattern recognition can be used for the quality control of the Tibetan medicine Ershiwuwei luxue pills.
期刊: 2021年第32卷第04期
作者: 钟镥,刘川,何青秀,张静,张艺
AUTHORS: ZHONG Lu,LIU Chuan ,HE Qingxiu ,ZHANG Jing,ZHANG Yi
关键字: 藏药二十五味驴血丸;高效液相色谱法;指纹图谱;主成分分析;聚类分析;含量测定
KEYWORDS: Tibetan medicine Ershiwuwei luxue pills ;HPLC;Fingerprint;Principle component analysis ;Cluster analysis ;
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