LC-MS/MS法测定毛曼陀罗中莨菪碱、东莨菪碱和山莨菪碱的含量

DETERMINATION OF HYOSCYAMINE,SCOPOLAMINE AND ANISODAMINE IN DATURA INNOXIA BY LC-MS/MS

目的建立LC-MS/MS测定毛曼陀罗中莨菪碱、东莨菪碱和山莨菪碱含量的方法。方法利用盐酸与浓氨水和三氯甲烷提取毛曼陀罗根部中的生物碱,以乙腈-(0.05%甲酸)水溶液=68:32为流动相;Agilent ZORBAX SB-C18(2.1 mm×100 mm,3.5μm)的色谱柱;柱温25℃;流速0.3 mL/min;进样量:2μL。结果莨菪碱、东莨菪碱和山莨菪碱在分别为1~200 ng/mL;1~1000 ng/mL;1~1000 ng/mL的范围内有良好的线性关系。线性方程依次为:Y=813.8904X+2165.3000(r^2=0.9922);Y=714.1221X+517.4311(r^2=0.9892);Y=872.9971X+1024.4247(r2=0.9952);平均加样回收率分别为97.17%、97.2%、92.68%,RSD分别为1.02%、1.31%、1.41%。通过对12批曼陀罗根中莨菪碱、东莨菪碱和山莨菪碱的测定,其浓度范围分别为1~20 ng/mL;1~18 ng/mL;1~19 ng/mL。结论该方法简单、灵敏度高、稳定性好可用于毛曼陀罗中莨菪碱、东莨菪碱和山莨菪碱含量的测定。

Objective:To establish a LC-MS/MS method for the determination of hyoscyamine,scopolamine and anisodamine in Datura innoxia.Methods:The alkaloids were extracted with hydrochloric acid,ammonium hydroxide and chloroform.The chromatographic column was Agilent ZORBAX SB-C18(2.1×100 mm,3.5μm),the moving phase was composed of acetonitrile and 0.05%formic acid solution with the ratio of 68:32,the column temperature was 25℃,the flow rate was 0.3 mL/min and the injection amount was 2μL.Results:The linear ranges of hyoscyamine,scopolamine and anisodamine were 1 ng/mL^200 ng/mL,1 ng/mL^1000 ng/mL and 1 ng/m^1000 ng/mL respectively.The linear equations were Y=813.8904 X+2165.3000(R^2=0.9922),Y=714.1221 X+517.4311(R^2=0.9892),Y=872.9971 X+1024.4247(R2=0.9952)respectively.The average recoveries were 97.17%,97.2%,92.68%,and RSD were 1.02%,1.31%and 1.41%respectively.Conclusion:The method is simple,sensitive and stable for the determination of hyoscyamine,scopolamine and anisodamine.