高效液相色谱-荧光检测流速梯度法测定去卵巢小鼠脑内单胺类神经递质

Measurement of Monoamine Transmitters in the Brains ofOvariectomized Mice Using High Performance LiquidChromatography with Fluorescence Detectionand Gradient Flow Rate Elution

建立了同时检测去卵巢(OVX)小鼠脑内的去甲肾上腺素(NE)、多巴胺(DA)、5 羟色胺(5 HT)3种神经递质的高效液相色谱方法。采用的色谱条件为:C18柱分离,荧光检测器检测,流动相为0 1mol/L的KH2PO4缓冲液 甲醇(体积比为9∶1),设置单泵流速梯度洗脱。方法的检测限(S/N=3)均在10nmol/L以下,绝对回收率均在80%以上,样品日内测定峰面积的相对标准偏差在3 03%以下。该方法具有检测灵敏度高、样品制备简便等优点,对小鼠脑内的3种单胺类递质的检测具有实用价值。

A method for the determination of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) in brains of ovariectomized mice was established by using high performance liquid chromatograph with a fluorescence detector, a C_(18) column and a gradient flow rate delivered by a single pump. The mobile phase was a mixture of 0.1 (mol/L) KH_2PO_4 and CH_3OH (9/1,v/v). Detection limits of the method were determined to be lower than 10 (nmol/L), absolute recoveries were above 80% and the relative standard deviations (RSDs) of intra-day were all within 3.03%.The method has the advantages of high sensitivity and of simple sample preparation. Short analysis time and better separation of samples were also obtained.