HPLC测定不同种蟾皮药材中不同部位蟾蜍噻咛的含量

Determination of Bufothionine in Different Parts from Two Species of Toad Skin Using HPLC

目的:对不同种蟾皮不同部位中蟾蜍噻咛的含量进行了比较研究。方法:采用高效液相色谱法测定了不同种蟾皮中不同部位蟾蜍噻咛的含量。样品中蟾蜍噻咛峰通过与对照品进行HPLC-DAD 3D及HPLC-MS-MS比对分析进行了确定。提取方法通过正交试验确定为50倍量50%甲醇超声60 min。样品分析选用了WondsilTM C18色谱柱(4.6 mm×250 mm,5μm),以乙腈-水(6∶94)为流动相,检测波长226 nm。结果:该方法具有良好的线性(r=0.999 7)、精密度(RSD,0.59%)、稳定性(RSD,0.72%)和重复性(RSD,1.56%);平均加样回收率为98.57%,RSD为2.03%,表明该方法准确可靠。经测定,中华大蟾蜍与黑框蟾蜍不同部位均含有蟾蜍噻咛成分,含量为0.07%～0.17%。结论:蟾蜍噻咛在不同种蟾皮药材中的不同部位含量差异不大。

Objective： To determine bufothionine in different parts of the dried toad skin from two species. Method： A quantification method was developed by HPLC with photodiode array （PDA） detection. Bufothionine was identified unambiguously by HPLC-DAD 3D and HPLC-MS-MS analysis in comparison with reference standard. The extraction method was optimized as 50 times 50% MeOH ultrasonic 60 min. The analysis was performed on WondsilTM C18 column （4.6 mm ~ 250 ram, 5 ixm） with ACN-HzO （6： 94） as mobile phase and detection wavelength was at 226 nm. Result： The samples were analyzed with good linear regression relationship （ r = 0. 999 7 ）, precisions （ RSD, 0.59% ）, repeatability （ RSD, 1.56% ）, stability （ RSD, 0.73% ） and recovery （98.57% ; RSD, 2.03% ）. Bufothionine was detected in different parts of toad skin from Bufobufo gargarizans cantor and Bufo melanostictus Schneider. The contents were from 0.07% to 0. 17%. Conclusion ： The content of bufothionine revealed no significant differences among the different parts in these two species of toad skin.