HPLC测定不同种蟾皮药材中不同部位蟾蜍噻咛的含量

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WondsiITM 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.

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

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 mm,5 μm) with ACN-H2O(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.