HPLC法测定紫花妇炎合剂中阿魏酸的含量

HPLC Method for Determining Content of Ferulic Acid in Zihua Fuyan Mixture

目的研究分析HPLC法测定紫花妇炎合剂中阿魏酸含量的方法。方法采用HPLC法,C18柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.085%磷酸溶液(17∶83);流速1.00 mL/min,柱温35℃,检测波长为316nm。系统适用性试验,分别精密吸取对照品溶液、供试品溶液和阴性对照溶液,利用上述2.1色谱条件,取10μL注入液相色谱仪,记录色谱图;线性关系的考察,精密量取2.2项下的对照品储备液1.0,2.0,5.0,10.0,20.0 mL,分置50 mL,量瓶中,用流动相准确稀释至刻度,摇匀。分别准确进样10μL,按2.1色谱条件,测定其峰面积,以对照品进样量(μg)为横坐标,以峰面积为纵坐标,进行线性回归,得阿魏酸的回归方程;精密度试验取对照品溶液,按照2.1的色谱条件,重复进样6次,记录峰面积;稳定性试验,取批号为20180411的供试品,按2.3项下方法制备供试品溶液,在室温下放置后,按照2.1的色谱条件,分别在0、2、4、8、12 h,取该溶液10μL注入液相色谱仪,测定峰面积;重复性试验,取批号20180411的供试品,按照2.3项下的操作,平行制备6份供试品溶液,照2.1的色谱条件,每份样品平行进样2次,10mL/次,测定峰面积。结果阿魏酸的测定不受其他杂质峰的干扰;阿魏酸在2.56~51.2μg/mL浓度范围内线性关系良好(r=0.9997),平均回收率97.3%;阿魏酸峰面积的RSD=0.76%。结论本实验建立了紫花妇炎合剂中阿魏酸的含量测定的分析方法,该方法的供试品溶液制备简单快捷,且所建立的测定方法经方法学考察,符合定量分析要求,操作简单,结果准确可靠,重现性好,可为紫花妇炎合剂控制指标的选择提供参考,为紫花妇炎合剂质量标准的建立提供一定的科学依据,有利于紫花妇炎合剂整体质量的控制和评价。

Objective To study the HPLC method for the determination of ferulic acid in Zihua Fuyan Mixture.Methods HPLC method was used.C18 column(250 mm×4.6 mm,5μm)was selected.The mobile phase was acetonitrile-0.085%phosphoric acid solution(17∶83).The flow rate was 1.00 mL/min and the column temperature was 35℃.The detection wavelength was 316 nm.For system suitability test,we accurately sucked the control solution,tested the solution and negative control solution respectively.Taking 10μL of the above chromatographic conditions,we injected it into the liquid chromatograph,and recorded the chromatogram.For the investigation of linear relationship,we accurately measured 1.0,2.0,5.0,10.0,20.0 mL of the control solution under 2.2,and placed it in a measuring bottle of 50 mL,accurately diluted it to the scale with mobile phase,and shook it up.We accurately injected 10μL respectively,and determined the peak area according to the 2.1 chromatographic conditions.We took the control sample injection amount(μg)as the abscissa and the peak area as the ordinate,and carried out linear regression to obtain the regression equation of ferulic acid.We took the control solution in the precision test,repeated the injection 6 times according to the 2.1 chromatographic conditions,and recorded the peak area.We took the test sample with the batch number of 20180411 in the stability test and prepared the test solution according to the method in 2.3,placed it at room temperature,and injected 10μL of the solution into the liquid chromatograph at 0,2,4,8 and 12 h respectively according to the chromatographic conditions in 2.1,and measured the peak area.For the repeatability test,we took the test solution in batch No.20180411,prepared 6 test solution in parallel according to the operation in 2.3,and injected each sample twice in parallel according to the chromatographic conditions in 2.1.The peak area was measured at10μL.Results The determination of ferulic acid was not interfered by other impurity peaks.The linear relationship of ferulic acid was good in the concentration range of 2.56-51.2μg/mL(r=0.9997).The average recovery was 97.3%,and the RSD of ferulic acid peak area was 0.76%.Conclusion The assay method of ferulic acid in Zihua Fuyan Mixture was established in this experiment.The preparation of the test solution was simple and fast.The assay method met the requirements of quantitative analysis and was simple,accurate and reliable with had good reproducibility.It can provide reference for the selection of control indexes of Zihua Fuyan Mixture and the quality of Zihua Fuyan Mixture.The establishment of the quantitative standard provides a scientific basis for the control and evaluation of the quality of Zihua Fuyan Mixture.