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

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.