LC-MS/MS法测定针刺穴位给药大鼠心肌组织中多巴酚丁胺的含量

目的: 建立液相色谱-串联质谱(LC-MS/MS)方法,对多巴酚丁胺在大鼠心肌组织分布的经时变化进行研究。 方法: 优化多巴酚丁胺及内标盐酸苯妥拉明离子化条件,开展方法学考察,以验证新建分析方法的准确性、精密度等,在此基础上将该方法用于比较分析肌肉注射和穴位注射给药大鼠心肌组织中多巴酚丁胺的含量。 结果: ESI(+)为多巴酚丁胺提供最佳的离子化条件,采用MRM工作模式, 用m/z 302.1→107.1来检测多巴酚丁胺,同时,以盐酸苯妥拉明m/z 256.3→167.1为内标化合物。多巴酚丁胺在3.75~240 ng·g^-1范围内线性良好,回归方程为Y=0.0053X+0.1956,r=0.9988,检测限为0.375 ng·g^-1,高、中、低浓度的准确度为95.2%~104.0%。日内精密度(n=6)为3.0%~7.1%,日间精密度(n=3)为4.0%~14.6%。 结论: 该方法对多巴酚丁胺测定具有良好的灵敏度、准确度、精密度以及专属性,完全适应于大鼠心肌组织中的多巴酚丁胺含量的测定,同时为针药联合干预后靶器官药物含量的比较以及针药联合干预后心肌组织中药物含量的检测等研究工作打下了重要的分析方法学基础。

Objective: To develop a sensitive and reliable method for determination of dobutamine in heart of rats. Method: The optimal ionization and fragmentation conditions,as well as liquid chromatographic ones,to detect dobutamine were developed and validated.The new developed analytical method was further applied in determination of dobutamine from rats heart after intramuscular injection and point injection of dobutamine. Result: The detection was performed by MRM mode via electrospray ionization(ESI)source operating in the positive ionization mode. The precursor-to-product ion transitions of the analyte dobutamine was m/z 302.1→107.1 with internal standard(IS,diphenhydramine hydrochloride)at m/z 256.3→167.1 The method was linear over the concentration range of 3.75-240 ng·g^-1(r=0.9988).The limit of detection was 0.375 ng·g^-1.The accuracy was 95.2%-104.0%.The intra-and inter-day precision values were 3.0%-7.1%(n=6)and 4.0%~14.6%(n=3),respectively.[HTH][STHZ][WTHZ]Conclusion:[HTSS][STBZ][WTBZ]The fully validated LC-MS/MS method has been successfully applied to determination of dobutamine in heart of rats.