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

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...

Fingerprint Establishment,Content Determination and Chemical Pattern Recognition Analysis of Tibetan Medicine Ershiwuwei Luxue Pills

OBJECTIVE:To establish fingerprint of Tibetan medicine Ershiwuwei 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 Ershiwuwei 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 Ershiwuwei 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 1.232-11.092μg/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),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 0.175-0.310,0.351-0.632,0.274-0.395,0.186-0.278,6.956-8.636 mg/g,respectively.Cluster analysis showed that 10 batches of Tibetan medicine Ershiwuwei 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.