高效液相色谱法测定不同产地湘莲中荷叶碱、莲心碱、芦丁的含量

Determination of nuciferine, liensinine, rutin in Hunan lotus seeds from different origins by high performance liquid chromatography

目的建立高效液相色谱法同时测定湘莲中荷叶碱、莲心碱、芦丁的方法。方法在酸性条件下超声提取3种待测组分。色谱条件如下:C_(18)(250 mm×4.6 mm,5μm)色谱柱;流动相:甲醇-0.1 mol/L乙酸铵(用乙二胺调节pH值至7.5~8.0)(55:45,V:V);流速:1.0 m L/min;检测波长:275 nm;柱温30℃;进样量10μL。结果荷叶碱、莲心碱、芦丁分别在7.636~152.7μg/mL(r^2=0.9999,n=5)、7.543~150.9μg/mL(r^2=0.9995,n=5)、7.543~150.9μg/mL(r^2=0.9998,n=5)范围内线性关系良好,加标回收率为96.9%~100.5%之间,相对标准偏差(relative standard deviation,RSD)为1.1%~1.4%。结论该法同时测定湘莲中荷叶碱、莲心碱、芦丁,简单快速,定量准确,可以为评价不同产地湘莲功效成分提供参考。

Objective To establish a method for simultaneous determination of nuciferine, liensinine and rutin in Hunan lotus seeds by high performance liquid chromatography. Methods Totally 3 kinds of components were extracted by ultrasonic under acid condition. The chromatographic conditions were as follows： column was C18 （250 mm×4.6 mm, 5 μm）, methanol-0.1 mol/L ammonium acetate （to adjust the pH value to 7.5~8.0 with ethylenediamine） （55：45, V：V） was as mobile phase, flow rate was 1 mL/min, detection wavelength was 275 nm, column temperature was 30 ℃ and sample size was 10 μL. Results The nuciferine, liensinine and rutin showed a good linear range at 7.636~152.7 μg/mL （r^2=0.9999, n=5）, 7.543~150.9 μg/mL （r^2=0.9995, n=5） and 7.543~150.9 μg/mL （r^2=0.9998, n=5）, respectively. The recoveries were 96.9%~100.5% at the spiked levels with relative standard deviation of 1.1%~1.4%. Conclusion This method is accurate, simple and reliable, and it provides a reference for the evaluation of different origin of lotus ingredients, which can be applied to the simultaneous determination of nuciferine, liensinine and rutin in lotus seeds.