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目的:建立小鼠脑内去甲肾上腺素含量的高效液相色谱-电化学(HPLC-ECD)测定方法。方法:小鼠脑组织采用70%高氯酸沉淀蛋白后,以HPLC-ECD法进行分析。使用汉邦lichrospher C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为醋酸钾缓冲液-甲醇(95:5,pH 4.5),流速为1.0 ml·min~(-1),温度为25℃;电化学检测器电压为0.6 V,电流为100 nA,频率为0.05 Hz。结果:去甲肾上腺素在0.1～25μmol·L~(-1)范围内线性关系良好,日内、日间RSD分别小于7.81%和3.61%,回收率为75.05%,测得小鼠脑内去甲肾上腺素含量为(8.94±0.71)μg·g~(-1)。结论:该方法快速、准确,适用于小鼠脑内去甲肾上腺素的测定
Objective: To establish a HPLC-ECD method for the determination of norepinephrine in mouse brain. Methods: The brain tissues of mice were analyzed by HPLC-ECD after 70% perchloric acid precipitated protein. The mobile phase consisted of Lichrospher C_ (18) column (250 mm × 4.6 mm, 5 μm) with potassium acetate acetate buffer (95: 5, pH 4.5) at a flow rate of 1.0 ml · min -1 The temperature was 25 ° C; the electrochemical detector voltage was 0.6 V, the current was 100 nA and the frequency was 0.05 Hz. Results: The linearity of norepinephrine in the range of 0.1 ~ 25μmol·L ~ (-1) was good. The intra-day and inter-day RSD were less than 7.81% and 3.61%, respectively. The recovery rate was 75.05% Epinephrine content was (8.94 ± 0.71) μg · g ~ (-1). Conclusion: The method is rapid and accurate and is suitable for the determination of norepinephrine in the brain of mice