高效液相色谱法同时测定摩罗丹中5种成分含量

Simultaneous Determination of Five Components in Moluo Dan by HPLC

目的 建立同时测定摩罗丹中5种成分含量的高效液相色谱法。方法 色谱柱为Agilent Eclipse XDB-C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.05%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,检测波长分别为210 nm(哈巴苷)、230 nm(芍药苷)、280 nm(延胡索乙素、哈巴俄苷)、203 nm(人参皂苷Rb1),柱温为20℃,进样量为10μL。结果 哈巴苷、芍药苷、延胡索乙素、哈巴俄苷、人参皂苷Rb1质量浓度分别在0.34~3.77μg/mL(r=0.999 4)、41.32~454.54μg/mL(r=1.000 0)、1.41~15.52μg/mL(r=0.999 8)、2.02~22.24μg/mL(r=0.999 7)、72.19~794.13μg/mL(r=0.999 9)范围内与峰面积线性关系良好(n=6);检测限分别为0.06,0.20,0.30,0.20,0.40μg/mL,定量限分别为0.17,0.62,0.70,0.50,1.25μg/mL;精密度、稳定性、重复性试验结果的RSD均小于2.0%;平均加样回收率分别为99.43%,99.85%,97.93%,98.61%,99.77%,RSD分别为1.38%,0.30%,2.04%,1.52%,0.53%(n=9)。结论 该方法操作简便、结果准确、重复性好,可用于同时测定摩罗丹中5种成分的含量。

Objective To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of five components in Moluo Dan.Methods The chromatographic column was Agilent Eclipse XDB-C_(18) column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.05%phosphoric acid aqueous solution(gradient elution),the flow rate was 1.0 mL/min,the detection wavelengths were 210 nm(harpagide),230 nm(paeoniflorin),280 nm(tetrahydropalmatine,harpagoside)and 203 nm(ginsenoside Rb1)respectively,the column temperature was 20℃,and the injection volume was 10μL.Results The linear ranges of harpagide,paeoniflorin,tetrahydropalmatine,harpagoside and ginsenoside Rb1 were 0.34-3.77μg/mL(r=0.9994,n=6),41.32-454.54μg/mL(r=1.0000,n=6),1.41-15.52μg/mL(r=0.9998,n=6),2.02-22.24μg/mL(r=0.9997,n=6),72.19-794.13μg/mL(r=0.9999,n=6)respectively.The limits of detection(LOD)of the above five components were 0.06,0.20,0.30,0.20,0.40μg/mL respectively,the limits of quantification(LOQ)of the above five components were 0.17,0.62,0.70,0.50,1.25μg/mL respectively.The RSDs of precision,stability and repeatability tests were all lower than 2.0%.The average recovery rates of the above five components were 99.43%,99.85%,97.93%,98.61%,99.77%,with RSDs of 1.38%,0.30%,2.04%,1.52%,0.53%(n=9)respectively.Conclusion This method is simple,accurate and repeatable,which can be used for the simultaneous determination of the contents of five components in Moluo Dan.