HPLC法测定草乌中新乌头碱、乌头碱和次乌头碱的含量

Determination of Mesaconitine,Aconitine and Hypaconitine in Radix Aconitum Kusnezoffii by HPLC

[目的]建立HPLC法测定草乌中新乌头碱、乌头碱和次乌头碱含量的方法。[方法]采用Agilent C18色谱柱(250mm×4.6mm,5μm);以甲醇-0.1%氨水(磷酸调pH9.0)梯度洗脱,检测波长235nm,柱温30℃,流速1.0mL/min。[结果]三种生物碱得到有效分离,线性范围分别为新乌头碱0.1944～1.1664μg(r=0.9997)、乌头碱0.9920～5.9520μg(r=0.9998)、次乌头碱0.0792～0.4752μg(r=0.9994);平均加样回收率分别为98.3%(RSD=0.97%,n=6)、99.2%(RSD=0.84%,n=6)、98.1%(RSD=1.1%,n=6)。[结论]该方法重复性好,可操作性强,能够用于对草乌的质量控制。

[ Objective ] To establish a method to determine mesaconitine, aconitine and hypaconitine in Radix Aconitum kusnezoffii by HPLC. [ Methods ] An Agilent C18 column （4.6 x 250mm, 5 μm） was used by a gradient elution using methanol -0. 1% ammonia （adjusted to pH9.0 with phosphate acid） as the mobile phase, the detection wavelength was 235 nm, the column temperatuer was 30℃ and the flow rate was 1.0 mL · min - 1. [ Results ] The method had effective separation of the three kinds of alkaloids. Mesaconitine, aconi- tine and hypaconitine showed a good linear relationship at the range of was 0. 1944 - 1. 1664μg（ r = 0. 9997 ） , 0. 9920 - 5. 9520txg（ r = 0. 9998 ） and 0. 0792 - 0. 47521μg（ r = 0. 9994 ）, respectively ; The average recovery was 98.3%（RSD=0.97%,n=6）, 99.2% （RSD=0.84%,n=6） and 98.1%（RSD=l.1%,n=6）, re- spectively. [ Conclusion ] The method is reproducible, easy to operate. It can be used for quality control of Ra- dix Aconitum kusnezoffii.