高效液相色谱法同时测定分心木中芦丁、槲皮素、金丝桃苷含量

Content Determination of Rutin,Quercetin and Hypericin in Diaphragma Juglandis Fructus by HPLC

目的建立同时测定分心木中芦丁、槲皮素、金丝桃苷含量的高效液相色谱法。方法色谱柱为Kromasil 100-5 C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.4%磷酸水溶液(梯度洗脱),流速为1.0 m L/min,检测波长为360 nm,柱温为25℃,进样量为10μL。结果芦丁、槲皮素、金丝桃苷线性范围分别为36.60~366.00μg/m L、48.10~481.00μg/m L、4.80~48.00μg/mL(r=0.999,n=5);平均加样回收率分别为100.10%,99.72%,100.02%,RSD分别为0.71%,0.18%,0.26%(n=9);精密度、稳定性、重复性试验的RSD均小于2%。结论该方法简便快捷,结果准确可靠,重复性和稳定性均较好,可用于同时测定分心木中芦丁、槲皮素、金丝桃苷的含量。

Objective To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of rutin,quercetin and hyperoside in Diaphragma Juglandis Fructus.Methods The chromatographic column was Kromasil 100-5 C18 column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.4%phosphoric acid aqueous solution(gradient elution),the flow rate was 1.0 m L/min,the detection wavelength was 360 nm,the column temperature was 25℃,and the injection volume was 10μL.Results The linear ranges of rutin,quercetin and hyperoside were 36.60-366.00μg/m L,48.10-481.00μg/m L,4.80-48.00μg/m L(r=0.999,n=5),respectively.The average recoveries of rutin,quercetin and hyperoside were 100.10%,99.72%and 100.02%with RSDs of 0.71%,0.18%and 0.26%(n=9),respectively.The RSDs of precision,repeatability and stability tests were lower than2%.Conclusion The method is simple,rapid,accurate,reliable,reproducible and stable,which can be used for the simultaneous determination of rutin,quercetin and hyperoside in Diaphragma Juglandis Fructus.