天仙子中消旋山莨菪碱、东莨菪碱和阿托品的含量测定及HPLC特征图谱研究

Contents of raceanisodamine,Scopolamine and hyoscyamine in henbane seed and its specific chromatogram by HPLC

目的:建立天仙子中消旋山莨菪碱、东莨菪碱和阿托品的含量测定方法及天仙子药材的HPLC特征图谱。方法:采用HPLC法对不同产地天仙子进行特征图谱研究,建立HPLC特征图谱共有模式,并进行相似度比较。色谱柱为资生堂CAPCELL-PAK C18(4.6 mm×250 mm,5μm),流动相为甲醇-乙腈-醋酸钠缓冲盐水溶液(5∶10∶85);流速1.0 ml/min;检测波长为210 nm;柱温:30℃。结果:消旋山莨菪碱、东莨菪碱及阿托品的线性范围分别为0.009 35～0.4676μg、0.054 63～2.7314μg和0.067 25～3.3623μg(r=0.9999);平均回收率分别为103.72%(RSD=1.72%)、98.57%(RSD=1.64%)和102.96%(RSD=1.26%);得到分离度较好的HPLC特征图谱,识别出5个特征共有峰。结论:本方法分离度较好,准确、可靠,为其质量控制提供了依据。

Objective： To develop an HPLC method for simultaneous determination of raceanisodamine, scopolamine and hyoscyamine in Henbane seeds and for study of specific chromatogram of Henbane seeds. Methods ： The analysis was carried out on an Shiseido Capcell -Pak C18 （250 mm x 4.6 mm,5 μm） column;The mobile phase consisted of 0.1% phosphoric acid aqueous （A） and 70% acetonitrile （B） with gradient elution （0 - 30 min ：60% A→35 % A ; 30 - 40 min ： 35 % A→30% A ;40 - 44 min ： 30% A→20% A;44-45 min：20% A→60% A;45 -60 min： 60% A）and the flow rate was 1.0 ml/min; the detection wavelength was 210 nm;the column temperature was 30 ℃. Result： The linear ranges of raceanisodamine, scopolamine and hyoscyamine were 0.009 35- 0.4676 μg ,0. 05463 - 2.7314 μg, and 0. 067 25 - 3. 3623 μg, respectively （ r 〉 0.9999 ） ; the average recoveries were 103.72% ,98.57% and 102.96%, respectively （RSD 〈 2% ,n = 6）. Conclusion： This method is simple and can be used to determine the above three compents with satisfactory accuracy and repeatability.