用反相离子对色谱-电化学检测法测定大鼠脑内单胺类物质

Assay Procedures for the Determination of Monoamines in Rat’s Brain by Means of Reversed-Phase Ion-Pair Chromatography with Electrochemical Detection

应用反相离子对色谱-电化学检测器测定大鼠脑NA、DOPAC、DA、HIAA、5-HT,35min内完成分析。本室对色谱条件进行了实验选择,发现4％乙腈能有效缩短样品保留时间和改善峰形,可取代流动相中甲醇;提高柱温至30℃也有助于分析时间的缩短;当pH=4.4,离子对D-樟脑-B-磺酸的浓度为11.5mmol/L时,分离效果最好。在9.766pg-500ng范围,线性关系的相关系数平均为0.9987±0.0012;平均回收率119％;前三个化合物峰高变异系数0.67±0.19％,后三个峰为2.76±1.02％,所有保留时间的平均变异系数0.33±0.10％。

The determination of noradrenaline, dopamine, 5-hydroxytryptamine and their acidic metabolites was carried out by reversed-phase ion-pair chromatography with electrochemical detection. This method was able to analyze all monoamines and their metabolites during 35 min, It was found that 4% of acetonitrile was better than methanol for shortening the retention time of monoamines in the samples and improve their peak-figure. The chromatographic conditions chosen were:H.5mmol/L of ion-pair, 30℃ of column temperature, pH=4.4,4% acetonitrile in the mobile phase. In the range of 9.766 Pg-500 ng, the linear ralation between the amounts and the peak height was good, r=0.9987±0.0012; the mean recovery could reach 119%;the mean coefficient of variation of peak height were measured:0.67±0.19%and 2.76±1.02%, and the C.V. of retention time of all compounds was 0.33±0.10%.