HPLC波长切换法同时测定强肝颗粒中8种活性成分的含量

目的:建立HPLC波长切换法同时测定强肝颗粒中(R,S)-告依春、绿原酸、芍药苷、阿魏酸、洋川芎内酯Ⅰ、藁本内酯、丹酚酸B、丹参酮ⅡA8个成分的含量。方法:采用Phenomenex Kinetex C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)-0.05%磷酸溶液(B),梯度洗脱,检测波长为245 nm[0~18 min检测(R,S)-告伊春]、320 nm(18~24 min检测绿原酸)、230 nm(24~28 min检测芍药苷)、286 nm(28~70 min检测阿魏酸、洋川芎内酯Ⅰ、藁本内酯、丹酚酸B和丹参酮ⅡA),流速1.0 ml·min^-1,柱温35℃,进样量10μl。结果:(R,S)-告依春、绿原酸、芍药苷、阿魏酸、洋川芎内酯Ⅰ、藁本内酯、丹酚酸B、丹参酮ⅡA质量浓度分别在9.04~180.80μg·ml^-1(r=0.9983)、20.40~408.00μg·ml^-1(r=0.9998)、13.06~261.20μg·ml^-1(r=0.9997)、10.90~218.00μg·ml^-1(r=0.9999)、7.68~153.60μg·ml^-1(r=0.9992)、11.24~224.80μg·ml^-1(r=0.9986)、4.02~80.40μg·ml^-1(r=0.9988)、7.60~152.00μg·ml^-1(r=0.9990)范围内与峰面积呈良好线性关系;平均加样回收率分别为97.6%,98.2%,99.1%,98.7%,96.4%,96.2%,97.3%,97.0%,其RSD分别为1.4%,1.0%,0.7%,1.0%,1.0%,1.5%,1.1%,1.9%。结论:该方法准确、简单、有效,可用于同时测定强肝颗粒中上述8种活性成分的含量。

Simultaneous Determination of Eight Constituents in Qianggan Granules by HPLC Wavelength Switching Method

Objective:To establish an HPLC wavelength switching method for the determination of(R,S)-goitrin,chlorogenic acid,paeoniflorin,ferulic acid,senkyunolideⅠ,ligustilide,salvianolic acid B and tanshinoneⅡA in Qianggan granules.Methods:The separation was performed on a Phenomenex Kinetex C18column(250 mm×4.6 mm,5μm)with gradient elution of acetonitrile(A)-0.05%phosphoric acid(B).The flow rate was 1.0 ml·min^-1 and the column temperature was 35℃.The injection volume was 10μl.The detection wavelength was set at 245 nm(0-18 min)for(R,S)-goitrin,320 nm(18-24 min)for chlorogenic acid,230 nm(24-28 min)for paeoniflorin and 286 nm(28-70 min)for ferulic acid,senkyunolideⅠ,ligustilide,salvianolic acid B and tanshinoneⅡA.Results:(R,S)-Goitrin,chlorogenic acid,paeoniflorin,ferulic acid,senkyunolideⅠ,ligustilide,salvianolic acid B and tanshinoneⅡA showed good linearity within the range of 9.04-180.80μg·ml^-1(r=0.9983),20.40-408.00μg·ml^-1(r=0.9998),13.06-261.20μg·ml^-1(r=0.9997),10.90-218.00μg·ml^-1(r=0.9999),7.68-153.60μg·ml^-1(r=0.9992),11.24-224.80μg·ml^-1(r=0.9986),4.02-80.40μg·ml^-1(r=0.9988),7.60-152.00μg·ml^-1(r=0.9990),respectively.The recovery of the 8 constituents was 97.6%,98.2%,99.1%,98.7%,96.4%,96.2%,97.3%and 97.0%with the RSD of 1.4%,1.0%,0.7%,1.0%,1.0%,1.5%,1.1%and 1.9%,respectively.Conclusion:The method is accurate,simple and effective,which can be used for the simultaneous determination of 8 active constituents in Qianggan granules.