高效液相色谱同时测定荷叶药材及其生物碱部位中4种生物碱的含量

HPLC determination of 2-hydroxy-1-methoxyaporphine,pronuciferine,nuciferine and roemerine in Nelumbo nucifera and its alkaloid fraction

目的:建立测定荷叶药材及其生物碱部位中4种生物碱类成分2-羟基-1-甲氧基阿朴啡、原荷叶碱、荷叶碱和莲碱含量的高效液相色谱方法。方法:采用Hypersil C_(18)柱(4.6 mm×250 mm,5μm),以乙腈-0.1%三乙胺水溶液为流动相,梯度洗脱,流速1.0 mL·min^(-1),柱温35℃,检测波长270 nm。结果:2-羟基-1-甲氧基阿朴啡、原荷叶碱、荷叶碱和莲碱的线性范围分别为0.110～0.658μg(r=0.999 5),0.021～0.126μg(r=0.999 5),0.103～0.618μg(r=0.999 8),0.086～0.514μg(r=0.999 5),荷叶生物碱部位平均加样回收率(n=6)分别为101.5%,99.14%,99.21%,98.41%;荷叶药材平均加样回收率(n=6)分别为99.53%,100.5%,97.51%,100.1%。结论:3批样品测定结果表明,该方法简便准确,可用于荷叶药材及其生物碱部位中4种生物碱类成分的含量测定。

Objective： To establish an HPLC method for the determination of four alkaloids, i. e. 2-hydroxy-l-methoxyapor-phine, pronuciferine, nuciferine and roemerine, in Nelumbo nucifera and its alkaloid fraction. Method： The determination was carried out at 35℃ on a Hypersil C18 column （4.6 mm× 250 mm, 5 μm）, eluting with acetonitrile-water containing 0. 1% triethylamine as mobile phases in gradient mode. The flow rate was 1.0 mL·min^-1 and detection at the wavelength was set at 270 nm. Result： The linear ranges of 2-hydroxy-1 -methoxyaporphine, pronuciferine, nuciferine and roemerine were 0. 110-0. 658 μg （r=0. 999 5 ）, 0. 021 0-0. 126 μg（r=0. 999 5 ）, 0. 103-0. 618 μg （ r = 0. 999 8 ）, 0. 085 6-0. 514 μg （ r = 0. 999 5 ）, with the average recoveries （ n = 6 ） were 101.5% , 99. 14% , 99. 21% and 98. 41% for the alkaloid fraction of N. nucifera and 99. 53%, 100. 5%, 97.51% and 100. 1% for N. nucifera respectively. Conclusion： The determination results of the three batches of samples showed that the method was easy and accurate which could be used to determine the contents of four components in N. nucifera and its alkaloid fraction.