液相色谱-质谱联用法测定小鼠脑组织中枸橼酸芬太尼浓度

Study on determination of fentanyl citrate in mice brain tissue by liquid chromatography-mass spectrometry

目的:建立测定小鼠脑组织中枸橼酸芬太尼的液相色谱-质谱联用法。方法:色谱柱为Agilent Zorbax SB-C18(2.1mm×150mm,3μm);流动相为乙腈-0.05%甲酸(38∶62),流速0.3mL·min-1;柱温35℃;质谱条件为电喷雾电离源(ESI),以选择性正离子方式检测,枸橼酸芬太尼和内标盐酸美金刚的选择检测离子分别为m/z337.30([M+H]+)、m/z180.20([M+H]+)。小鼠尾静脉注射给予枸橼酸芬太尼(0.1mg·kg-1)30min后,测定12只小鼠脑组织内芬太尼的浓度。结果:小鼠脑组织内枸橼酸芬太尼的质量浓度为(18.4±6.4)μg·L-1,线性范围为1～50μg·L-1,检测限为0.25μg·L-1,方法回收率均大于95%。结论:本方法专属性强,灵敏度高,线性关系良好,操作简便,适用于小鼠脑组织内枸酸橼芬太尼浓度的测定。同时也可以为其他生物组织样本中芬太尼浓度测定提供参考。

OBJECTIVE To develop a sensitive and specific LC/MS method for determination of fentanyl citrate in mice brain tissue. METHODS The LC/MS method was performed with Agilent Zorbax Eclipse SB-C18 （2. 1 mm × 150 mm, 3 μm）. The mobile phase consisted of CH3CN and 0. 05% HCOOH （38：62）, Isocrtic elution was used. Analyses were run at flow rate of 0. 3 mL.min^-1. The temperature of column was 35℃. AP-ESI was applied and operated in positive mode. The ion of monitor：m/z 337. 30（fentanyl citrate） ,m/z 180. 20（memantadine hydrochoride, internal standard）. The mice brain tissue samples were obtained after injecting of 0. 1 mg.kg^-1 from the tail vein. RESULTS The linear calibration curve was observed in the concentration range of 1 50 μg. L^-1. The LOD and LOQ were 0. 25 μg L ^-1 and 1μg L^-1 , respectively. The relative recoveries were more than 95% ,and the inter-and intra-day precision （RSD） was below 5.0%. CONCLUSION The method was proved to be convenient,sensitive and rapid,and suitable for concentration study of fentanyl citrate in mice＇s brain.