HPLC法测定芍归活血益肝丸中芍药苷含量方法的研究

Determination of Paeoniflorin in Shaogui Huoxue Yigan Pills by HPLC

目的:用HPLC法测定芍归活血益肝丸中芍药苷的含量。方法:采用高效液相色谱法(HPLC),流动相是乙腈-0.05%磷酸(V乙腈∶V_(0.05%磷酸)=17∶83),Agilent TC-C_(18)(4.6 nm×250 nm,5.0μm)为色谱柱,柱温为40℃,流速为1.0 mL/min,检测波长为230 nm,进样量为10μL。结果:芍药苷的进样量与峰面积在0.6265~3.1324μg/mL范围内有良好的线性关系,线性回归方程y=437393x-26315,相关系数r=1(n=6),该法专属性良好,精密度、稳定性、重复性较高,加样回收率为96.23%,精密度为0.97%。结论:通过对三批样品的含量测定,暂定芍归活血益肝丸含白芍以芍药苷(C_(23)H_(28)O_(11))计剂量应不少于30.0 mg。验证表明本方法操作简单、快速、灵敏度高,具有良好的准确性和重现性,能够准确分析芍归活血益肝丸中芍药苷的含量,可以作芍归活血益肝丸中芍药苷的定性定量方法。

Objective:Determination of paeoniflorin content in Shao Gui activating blood and benefitting liver pills by HPLC method.Methods:Utilizing High-Performance Liquid Chromatography(HPLC),the mobile is composed of acetonitrile and 0.05%phosphoric acid(17∶83,volume ratio).An Agilent TC-C_(18)column(4.6 nm×250 nm,5.0μm)is used as the chromatography column.The column temperature is maintained at 40℃.The flow rate is set to 1.0 mL/min,and the detection wavelength is 230 nm.The injection volume for the samples is 10μL.Resμlts:A good linear relationship is observed between the sample volume of paeoniflorin and the peak area within the range of 0.6265 to 3.1324μg/mL.Linear regression equation y=437393x-26315,correlation coefficient r=1(n=6).This method has good specificity,precision,stability,and high repeatability.The recovery rate of sample addition is 96.23%,and the precision is 0.97%.Conclμsion:By measuring the content of three batches of samples,the tentative dosage of Shaogui Huoxue Yigan Wan containing white peony and paeoniflorin(C_(23)H_(28)O_(11))should not be less than 30.0 mg.Verification shows that this method is simple,fast,highly sensitive,and has good accuracy and reproducibility,Being able to accurately analyze the content of paeoniflorin in Shaogui Huoxue Yigan Wan,and can be used as a qualitative and quantitative method for paeoniflorin in Shaogui Huoxue Yigan Wan.