HPLC-MS/MS法测定超声提取刺五加果实中刺五加苷B、E和异嗪皮啶的含量  

Determination of Eleutheroside B,E and Isofraxidin in Fruit of Radix et Caulis Acanthopanacis Senticosi by HPLC-MS/MS with Ultrasonic-Assisted Extraction

作　　者：张琳[1,2,3] 刘洋[1,2,3] 盖庆辉 祖元刚 

机构地区：[1]东北林业大学生物资源生态利用国家地方联合工程实验室 [2]东北林业大学森林植物生态学教育部重点实验室 [3]东北林业大学林业生物制剂教育部工程研究中心,黑龙江哈尔滨150040

出　　处：《上海中医药大学学报》2014年第4期90-95,共6页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家"十二五"科技支撑项目(2011BAD33B0203)

摘　　要：目的：建立一个高效液相-质谱联用（HPLC-MS/MS）测定刺五加果实中刺五加苷B、E和异嗪皮啶含量的方法。方法：建立HPLC-MS/MS方法,进行色谱和质谱方法的考察,得到最优的分析条件,即流动相为乙腈-0.05%的甲酸溶液（20∶80,V/V）,流速为0.5 ml/min。采用电喷雾离子源（ESI）进行电离,多离子反应监测（MRM）模式进行信号采集,采用外标法进行定量;采用HPLC-MS/MS方法同时测定刺五加果实中刺五加苷B、E和异嗪皮啶的含量。结果：刺五加苷B、刺五加苷E和异嗪皮啶分别在0.7~30 000 ng/ml、0.4~10 000 ng/ml和0.3~16 000 ng/ml浓度范围内呈良好的线性关系（r〉0.999 7）,检出限（LOD）分别为0.17 ng/ml、0.12 ng/ml和0.09 ng/ml,定量限（LOQ）分别为0.7 ng/ml、0.4 ng/ml和0.3 ng/ml,不同浓度下的平均加标回收率为98.10%、98.19%和98.21%;在最优提取条件下刺五加果实中刺五加苷B的含量为（1.935 0±0.001 5）mg/g,刺五加苷E的含量为（0.273 0±0.003 5）mg/g,异嗪皮啶的含量为（0.013 0±0.001 2）mg/g。结论：HPLC-MS/MS法灵敏度高、选择性好、分析时间短,可对刺五加果实中刺五加苷B、刺五加苷E和异嗪皮啶进行准确的定量分析。Objective： To establish a method for the determination of eleutheroside B,eleutheroside E and isofraxidin by HPLC-MS /MS method. Methods： A HPLC-MS /MS method was established for the simultaneous determination of eleutheroside B,eleutheroside E and isofraxidin in Radix et Caulis Acanthopanacis Senticosi. After established the HPLC-MS /MS method,we examined the chromatography and mass spectrometry methods. The optimum analytical conditions： mobile phase of acetonitrile-0. 05% formic acid（ 20： 80,V /V）; flow rate,0. 5ml /min,using electrospray ionization（ ESI） ionization,multiple reaction monitoring（ MRM） mode for signal acquisition,quantified using the external standard method. The contents of eleutheroside B,eleutheroside E and isofraxidin in the fruit of Radix et Caulis Acanthopanacis Senticosi was determined by HPLC-MS /MS. Results： The experimental results showed that： eleutheroside B,eleutheroside E and isofraxidin set at0. 7-30000 ng /ml,0. 4-10000 ng /ml and 0. 3-16000 ng /ml concentration range showed an excellent linear relationship（ r 0.9997）,respectively,the corresponding limits of detection（ LOD） were 0.17 ng/ml,0.12 ng/ml and 0.09 ng/ml.The limits of quantification（ LOQ） were 0. 7 ng /ml,0. 4 ng /ml and 0. 3 ng /ml. The average recoveries of different concentrations were 98. 10%,98. 19% and 98. 21%. Under optimal extraction conditions,the contents of eleutheroside B,eleutheroside E and isofraxidin were（ 1. 9350 ± 0. 0015）,（ 0. 273 ± 0. 0035） and（ 0. 0130 ± 0. 0012） mg /g as determined by the HPLC-MS /MS assay. Conclusion： This method has high sensitivity,better selectivity and shorter analysis time for accurate quantitative analysis of eleutheroside B,eleutheroside E and isofraxidin in Radix et Caulis Acanthopanacis Senticosi.

关 键 词：高效液相色谱-串联质谱法(HPLC-MS MS) 刺五加果实 刺五加苷B 刺五加苷E 异嗪皮啶 

分 类 号：O657.63[理学—分析化学] TS255.7[理学—化学]