UPLC法同时测定草乌中6种单酯和双酯型生物碱

Determination of Six Kinds of Monoester and Diester- alkaloids in Radix Aconiti Kusnezoffii by UPLC

目的：建立草乌中单酯及双酯型生物碱成分的超高效液相色谱定量方法。方法：草乌粉末用氨试液浸润2 h后,以异丙醇-醋酸乙酯（1∶1）混合液浸泡12 h,超声处理30 min,提取液蒸干后溶于乙腈中,经微孔滤膜滤过后进行UPLC分析。UPLC分析采用BEH shield RP18（2.1 mm×50 mm,1.7μm）色谱柱,以乙腈（0.2%三乙胺）-水为流动相进行梯度洗脱,流速0.4 m L·min-1,检测波长为235 nm,柱温35℃。结果：草乌中6种单酯型和双酯型生物碱分离良好,各成分含量与峰面积在测定范围内均呈良好的线性关系（r〉0.999）,精密度高,重现性好,平均加样回收率为98.64%~100.32%,6 min完成1次进样测定,工作效率高。结论：建立UPLC同时测定草乌中6种单酯型和双酯型生物碱含量的方法,该方法稳定准确,简便快速,为草乌药材质量控制及进一步开发利用奠定基础。

Objective： To establish a UPLC method for simultaneous determination of monoester and diester aconitium alkaloids. Method： The powder of Radix aconiti kusnezoffii was infiltrated by adding ammonia solution for two hours,and then was soaked with a mixture solvent of isopropanol- ethylacetate（ 1 ∶ 1） for twelve hours and extracted by ultrasonic way for 30 min. The extract solution was evaporated to dryness and dissolve in acetonitrile. The sample solution was filtered with microfiltration membrane and analyzed by UPLC. The assay was performed on BEH shield RP18（ 2. 1 mm × 50 mm,1. 7 μm） and eluted with a mobile phase consisted of acetonitrile（ 0. 2% triethylamine）- ultrapure water. The flow rate was 0. 4 m L·min- 1and monitored by a UV detector at 235 nm. The temperature of chromatographic column was 35℃. Result： The six kinds of monoester and diesteralkaloids in radix aconiti kusnezoffii were separated very well. The linear relation of aconitum alkaloids content and the peak area were excellent over the studied concentration ranges（ r 〉0.999）. The accuracy of the method was high and the reproducibility was fine. The extraction recovery was ranged from 98.64% to 100. 32% for the six alkaloids. The one sample was only analyzed for six minutes and the work was efficient.Conclution： The UPLC method for simultaneous determination of monoester and diester- aconitum alkaloids was stable and accurate,simple and quick. It can be used for quality control of radix aconiti kusnezoffii and can lay a foundation of its further development and utilization.