中药饮片制川乌、制草乌中乌头碱、次乌头碱的HPLC测定

HPLC Determination of Aconitine and Hypaconitine in Prepared Chuan Wu and Cao Wu

目的:建立一种测定中药饮片制川乌、制草乌中乌头碱、次乌头碱含量的高效液相色谱方法。方法:饮片粉碎成粗粉,用氨试液润湿,乙醚浸提14h,提取液挥弃乙醚,残渣以二氯甲烷溶解,定容至5mL,用0.01mol·L^(-1)H_2SO_4溶液1mL萃取等体积的二氯甲烷液,取水相进样20μL,以Kromasil C_(18)(250mm×4.6mm,5μm)为色谱柱,甲醇-水-氯仿-三乙胺(70:30:2:0.1)为流动相,检测波长230nm,柱温为室温。结果:乌头碱在9.63-96.26μg·mL^(-1),次乌头碱在24.2-242μg·mL^(-1)范围内,线性关系良好(进样量为20μL),方法加样回收率>96%,RSD<2%。结论:该方法准确,精密度高,分离效果好,操作简单。

Objective: To develop an HPLC method for assay of aconitine and hypaconitine in prepared pieces of Chuan Wu and Cao Wu. Methods: The prepared pieces were milled into ChP coarse powders and then moistened with ammonia test solution, extracted with ether by maceration for 14h. The extract was evaporated to dryness, then, the residue was transferred to a 5-mL volumetric flask and diluted to volume with dichloromethane. 1mL of the dichloromethane solution was extracted with equal volume of 0. 01mol·L^(-1) sulfuric acid solution,then,20μL of water phase was analyzed by HPLC using a C_(18) column, methanol-water-chloroform-triethylamine(70: 30:2:0. 1) as mobile phase and UV detection at 230nm. Results: The calibration curves of aconitine and hypaconitine were linear, respectively, from 9. 63 to 96. 26μg·mL^(-1) and from 24. 2 to 242μg·mL^(-1), with injection volume of 20μL. The recoveries of this method were over 96% with RSD less than 2%. Conclusion: The method is accurate, precise, simple and with good resolution.