大鼠血浆中辣椒素的HPLC-MS/MS测定

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

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100μl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。

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.