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Simultaneous determination of the content of danshensu,puerarin,daidzin and salvianolic acid B in Bushen Huoxue prescription by HPLC wavelength switching technology

WANG Zi-yan;LI Chun-qin;MU Min-jie;DONG Jia-yue;LIU Mei-lin;ZHANG Mei;XIE Xue-jun;School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Key Laboratory of Standardization of Chinese Herbal Medicines of Ministry of Education,State Key Laboratory Breeding Base of Systematic research,Development and Utilization of Chinese Medicine Resources;School of Clinical Medicine,Chengdu University of Traditional Chinese Medicine;  
Objective:To develop an HPLC method for simultaneously determination of 4 components including danshensu,puerarin,daidzin and salvianolic acid B in Bushen Huoxue prescription.Methods:The Inertsil ODS-SP C_(18)(4.6 mm×250 mm,5 μm)was adopted.The mobile phase consisted of acetonitrile(A)-0.2% aqueous phosphoric acid(B)in a linear gradient elution mode.The column temperature was 30 ℃.The detection wavelength was set at 282 nm for danshensu in the first 12 min,then changed to 250 nm for puerarin and daidzin between 13 and 22 min,and changed to 286 nm for salvianolic acid B in the final 14 min.Results:The linear ranges of danshensu,puerarin,daidzin and salvianolic acid B fell within the ranges of 0.205 6-2.056 μg(r=0.999 7),0.1-1.0 μg(r=0.999 6),0.050 1-0.501 μg(r=0.999 4),1.0-10.0 μg(r=0.999 5),respectively.The recoveries(n=6)of danshensu,puerarin,daidzin and salvianolic acid B were 99.3%,100.7%,99.3%,98.8%,and the relative standard deviations were 1.1%,1.6%,1.6%,1.6%,respectively.The results of content assay(n=3)were 3.328 4-4.707 0,0.919 6-1.631 4,0.455 1-0.610 8,29.478 4-31.116 9 mg·g~(-1),separately,and the relative standard deviations were 1.6%-2.2%,0.4%-1.1%,1.2%-2.0%,0.8%-1.3%,respectively.Conclusion:This established method was proved by methodology validation that it can provide a reference for quality standard of Bushen Huoxue prescription.
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