高效液相色谱法测定阿尔茨海默病患者血浆中4种单胺类神经递质

目的基于高效液相色谱建立一种同时检测阿尔茨海默病患者血浆中去甲肾上腺素(NE)、肾上腺素(E)、多巴胺(DA)、5-羟色胺(5-HT)含量的方法。方法采用C18反相色谱柱,流动相:甲醇∶水相(0.1mol/L磷酸二氢钾)=3∶97,pH=3.7。流速:0.8ml/min,荧光检测器检测,激发波长:290nm,发射波长:330nm,柱温:室温。结果检测限:NE为0.02nmol/ml、E为0.01nmol/ml、DA为0.04nmol/ml、5-HT为0.02nmol/ml,4种神经递质RSD在2.2%～4.4%;回收率在82.9%～93.6%。线性范围NE为0.02～2.0nmol/ml、E为0.01～3.3nmol/ml、DA为0.04～3.6nmol/ml、5-HT为0.02～1.6nmol/ml,相关系数均在0.9977～0.9985之间。结论该方法操作简单、灵敏、重现性好,适用于阿尔茨海默病患者血液中单胺类神经递质水平研究。

Determination of kinds of monoamine neurotransmitters in Alzheimer' s blood plasma by high performance liquid chromatography

Objective A method detecting norepinephrine,epinephrine,dopamineand serotonin in Alzheimer's blood plasma by high performance liquid chromatography was developed.Methods C18 column was used,the mobile phase was composed of methanol-water(0.1mol/ml KH2PO4)(3∶97),pH 3.7.The flow rate was 0.8ml/min,fluorescence detection wave length λex was 290nm and λem was 330nm,column temperature was room temperature.Results The detection limits were 0.02nmol/ml for NE,0.01nmol/ml for E,0.04nmol/ml for DA,and 0.02nmol/ml for 5-HT.RSD was in the range of 2.2%-4.4%.The recoveries of all analytes were in the range of 82.9%-93.6%.The linear range was 0.02-2.0nmol/ml for NE,0.01-3.3nmol/ml for E,0.04-3.6nmol/ml for DA,and 0.02-1.6nmol/ml for 5-HT.The related coefficient was in the range of 0.9977-0.9985.Conclusion The assay had been applied successfully to measure the contents of these four monoamines in blood plasma of the Alzheimer's patients.