摘要
目的优化小鼠脑组织中单胺类神经递质多巴胺（DA）和5-羟色胺（5-HT）检测的实验条件,简化实验方法,提高检测方法的灵敏度。方法采用反相高效液相色谱法测定DA和5-HT,等度洗脱,流动相为：0.1 mol/L NaAc-甲醇（95∶5,v/v）（pH=6.0）,荧光检测器检测,保留时间定性,外标法定量。结果在优化的实验条件下,DA和5-HT在5μg/L!1000μg/L浓度范围内线性关系良好,相关系数分别为0.9989和0.9996;加标回收率分别为95.2%-100.4%和82.4%-100.6%;DA的日内和日间精密度分别为2.6%-9.0%和1.8%-8.7%;5-HT的日内和日间精密度分别为2.1%-6.2%和2.2%-10.7%。结论该方法简便、准确、灵敏,适用于小鼠脑组织中DA和5-HT的测定。
Objective To optimize the experiment conditions for the determination of dopamine （DA） and 5 - hydroxytryptamin （ 5 - HT） in mouse brain, simplify the experiment method and improve the test sensitivity. Methods DA and 5 - HT were determined by high performance liquid chromatography （HPLC） equipped with fluorescence detector using isocratic elution, and 0.1 mol/L sodium acetate buffer solution containing 0.1 mmol/L Na2 EDTA ： methanol （95：5, v/v） （pH = 6. O） was used as mobile phase. Retention time was used for qualitative analysis and external standard method was used for quantitative analysis. Results Using the optimized experimental conditions, the linearity of DA and 5 - HT were both well in the range of 5 μg/L -1000 μg/L and the linear correlation coefficients were 0.9989 and 0. 9996, respectively. The recoveries of DA and 5 - HT were 95.2% - 100.4% and 82.4% - 100.6%, respectively. The intra - day and inter - day precision of DA were 2.6% - 9.0% and 1.8% - 8.7%, respectively, while those of 5 - HT were 2.1% -.6.2% and 2.2% - 10.7%, respectively. Conclusion The method was simple, accurate, sensitive, and suitable for the simultaneous determination of the contents of DA and 5 - HT in mouse brain.
出处
《中国卫生检验杂志》
北大核心
2014年第14期1996-1999,共4页
Chinese Journal of Health Laboratory Technology
关键词
多巴胺
5-羟色胺
高效液相色谱
Dopamine
5 -hydroxytryptamin
High performance liquid chromatography
