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[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] The research aimed to establish a HPLC method for the determination of mesaconitine, aconitine and hypaconitine in Radix Aconitum. [Method] With Agilent C18 column (250mm × 4.6mm, 5μm) as eluent, the mobile phase was eluted with methanol - 0.1% aqueous ammonia (adjusted to pH 9.0 with phosphoric acid). The detection wavelength was set at 235nm and the column temperature was set at 30 ℃ with a flow rate of 1.0mL / min. [Result] The three alkaloids were effectively separated. The linear range was from 0.1944 to 1.1664μg of mesaconitine (r = 0.9997), from 0.9920 to 5.9520μg of aconitine (r = 0.9998), from 0.0792 to 0.4752μg of hypaconitine (r = 0.9994). The average recoveries were 98.3% (RSD = 0.97%, n = 6), 99.2% (RSD = 0.84%, n = 6) and 98.1% ). [Conclusion] This method has good repeatability and operability and can be used for the quality control of Radix.