南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (08): 1165-.

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大鼠血浆中辣椒素的HPLC-MS/MS测定

段炼,晏媛,刘媛,刘思佳,李国锋   

  • 出版日期:2013-08-20 发布日期:2013-08-20

Analysis of capsaicin in rat plasma using liquid chromatography-tandem mass spectrometry

  • Online:2013-08-20 Published:2013-08-20

摘要: 目的建立一种简单方便的血浆中辣椒素浓度的液质联用检测方法(HPLC-MS/MS)。方法血浆样本100 μl加入终浓度
为50 ng/ml的维拉帕米(内标),用混合溶剂乙酸乙酯∶丙酮(85∶15)萃取后用安捷伦6460三重四级杆质谱系统进行分析。采用
多反应监测(MRM)扫描方式,在ESI源正离子模式下,选择m/z 306→137作为辣椒素监测离子,m/z455→165为内标物监测离
子。结果样品分析时间仅需1.5 min,方法线性范围在1.85~370 ng/ml(r=0.9997),最低检测限为1.85 ng/ml。辣椒素在三个质
控浓度(3.7、37、370 ng/ml)检测下的提取回收率为77.34%,70.64%及78.02%。结论HPLC-MS/MS灵敏、快速、准确,可应用于
检测血浆中存在的微量辣椒素。

Abstract: Objective To establish a simple high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS)
system for determination of capsaicin in rat plasma. Methods Rat plasma (100 μl) was extracted with mixed solvent (ethyl
acetate∶acetone 85∶15) after addition of 50 ng/ml internal standard, and analyzed by Agilent 6460 Triple Quadrupole LC/MS
system. The detection was performed with multiple reactions monitoring using electrospray ionization. The precursor/product
ion transitions were monitored at m/z 306→137 (+Ion mode) for capsaicin. Verapamil was used as the internal standard at m/
z455→165. Results The analysis time was only 1.5 min in the positive mode; the calibration curve was linear in the
concentration range of 1.85-370 ng/ml. The lowest limit of quantification reached 1.85 ng/ml. The extraction recoveries were
77.34%, 70.64% and 78.02% for the three quality control concentration levels (3.7, 37, and 370 ng/ml), respectively. Conclusion
The LC-MS/MS system is simple, accurate, reliable and time-saving for determination of trace amount of capsaicin in plasma.