2种HPLC检测器测定莲子心中3种酚性生物碱含量的比较

Comparison of Three Phenolic Alkaloids' Content inPlumula Nelumbini by Using Two HPLC Detectors

目的建立反相HPLC-荧光检测(fluorescence detector,FD)法,测定中药莲子心中莲心碱(liensinine,Lien)、异莲心碱(isoliensinine,Iso)和甲基莲心碱(neferine,Nef)等3种酚性生物碱的含量,并与HPLC-紫外检测器(ultraviolet-vis-ible detector,UVD)的测定结果进行比较。方法采用Phecda C18色谱柱(5μm,4.6 mm×250 mm),乙腈-0.015 mol/L十二烷基磺酸钠-冰醋酸(55∶45∶1)为流动相;流速:1.0 mL/min;柱温:30℃。FD的激发波长为280 nm,发射波长为319 nm;UVD的检测波长为282 nm。结果用FD检测莲子心中3种酚性生物碱的回归方程和相关系数(r)分别为:ALien=134 994CLien+970 790,rLien=0.999 0;AIso=330 030CIso+647 307,rIso=0.999 3;ANef=165 207CNef+807 442,rNef=0.999 5。三者均具有良好的加样回收率(>95%),且所得莲子心中Lien、Iso和Nef的含量与UVD测得值基本一致。结论 HPLC-FD法测定莲子心中莲心碱、异莲心碱和甲基莲心碱含量方法可靠、结果准确,与HPLC-UVD法一样适用于莲子心药材中酚性生物碱的定量控制。

OBJECTIVE To establish a RP-HPLC method to determine the contents of three phenolic alkaloids,liensinine（Lien）,isoliensinine（Iso） and neferine（Nef） in Plumula Nelumbini（the green seed embryo of Nelumbo nucifera Gaertn） with the usage of fluorescence detector（FD）,and compare it with the same method by using ultraviolet-visible detector（UVD）.METHODS The analysis was performed on a column（5 μm,4.6 mm×250 mm） applying Phecda C18 as loading materials.The mobile phase was a mixture of acetonitrile-0.015 mol·L-1 aqueous solution of alkyl sodium sulfonate-acetic acid（55∶45∶1）.The volumetric flow rate was at 1.0 mL·min-1,and the column temperature was at 30 ℃.When using FD,the excitation wavelength was 280 nm and emission wavelength 319 nm;while the detective wavelength was 282 nm by using UVD.RESULTS There were good linearity relationships for Lien,Iso and Nef by using HPLC-FD,and their regression equations and correlation coefficients（r） were as follows：ALien =134994CLien ＋970790,rLien = 0.9990;AIso =330030CIso ＋647307,rIso = 0.9993;ANef =165207CNef ＋807442,rNef =0.9995.The average recoveries of the three alkaloids were all more than 95%.FD measuring data of Lien,Iso and Nef were almost kept the same as the ones by using UVD.CONCLUSION Just like HPLC-UVD,FD is of good accuracy,specificity and reproducibility in application of quantitative analysis of liensinine,isoliensinine and neferine in Plumula Nelumbini.