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建立了一种微透析活体取样与高效液相色谱-电化学检测法联用技术测定自由活动大鼠脑中7种单胺类神经递质的方法。高效液相色谱采用AglientXDB-C18柱,流动相为pH 3.0的0.1 mol/L H3PO4-NaH2PO4缓冲液与甲醇的混合液(90:10,V/V),流速为0.3 mL/m in,电化学检测的工作电极为玻碳电极,工作电位为0.55V。去甲肾上腺素、肾上腺素、3,4-二羟基苯乙酸、多巴胺、5-羟吲哚乙酸和5-羟色胺的线性范围为1.0～5.0×103ng/mL,高香草酸的线性范围为10.0～1.0×104ng/mL;7种物质相关系数均大于0.999,检出限在0.01 ng/mL水平。结合微透析活体取样,成功测定了自由活动大鼠脑纹状体中的神经递质,结果表明帕金森模型组大鼠脑中7种单胺类神经递质的含量较正常对照组有所降低。
A microdialysis in vivo sampling and high performance liquid chromatography - electrochemical detection method for the determination of free-moving rat brain monoamine neurotransmitters method. High performance liquid chromatography with AglientXDB-C18 column with a mobile phase of 0.1 mol / L H3PO4-NaH2PO4 buffer and methanol (90:10, V / V) at a flow rate of 0.3 mL / Detection of the working electrode for the glassy carbon electrode, the working potential of 0.55V. The linear range of norepinephrine, epinephrine, 3,4-dihydroxyphenylacetic acid, dopamine, 5-oxoindoleacetic acid and serotonin was 1.0-5.0 × 103ng / mL. The linear range of homovanillic acid was 10.0 ~ 1.0 × 104ng / mL. The correlation coefficients of seven substances were all above 0.999, with the detection limit of 0.01 ng / mL. Combined with microdialysis sampling in vivo, the neurotransmitter in the free striatum of the striatum of rats was successfully measured. The results showed that the content of 7 monoamine neurotransmitters in the brain of Parkinson’s model group was lower than that of the normal control group .