Simultaneous determination of biogenic monoamines in rat brain dialysates using capillary high-performance liquid chromatography with photoluminescence following electron transfer
Simultaneous determination of biogenic monoamines in rat brain dialysates using capillary high-performance liquid chromatography with photoluminescence following electron transfer
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DOI：
10.1021/ac051183g
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发表时间：
2006-03-15
影响因子：
7.4
通讯作者：
Weber, SG
中科院分区：
文献类型：
--
作者：
Jung, MC;Shi, GY;Weber, SG
同时测定脑内生物源性单胺，如多巴胺、血清素和3-甲氧基酪胺，对了解神经递质活性具有重要意义。本研究采用毛细管高效液相色谱-电子转移后光致发光检测技术对大鼠脑纹状体微透析液中生物源性单胺进行了灵敏测定。通过改变离子作用剂的浓度和有机改性剂的掺量来优化分离条件。高浓度离子相互作用剂使胺类化合物能够与干扰酸分离，但也使分离时间很长。为缩短分离时间，采用10% （v/v）乙腈作为有机改性剂。在3小时内分析了8个色谱运行的保留时间、峰高和峰宽。色谱图的重现性非常好，在3小时内峰高的变化小于1%。在最佳分离条件下，代谢酸的典型浓度检测限小于100 pM，单胺的浓度检测限与200 pM相似。样品进样量为500nl。因此，代谢酸的质量检测限小于50 amol，单胺的质量检测限接近100 amol。典型分离时间小于10 min。为了验证该技术，采用分离方法观察大鼠脑微透析样品中单胺浓度的药物诱导变化。麻醉大鼠纹状体局部灌注河豚毒素（一种钠通道阻滞剂）可降低透析液中多巴胺、3-甲氧基酪胺和血清素浓度。在7.7 min的时间分辨率下对纹状体透析液的连续监测显示，注射诺非芬可短暂增加大鼠脑透析液中多巴胺和3-甲氧基乳清的浓度。
Simultaneous determination of biogenic monoamines such as dopamine, serotonin, and 3-methoxytyramine in brain is important in understanding neurotransmitter activity. This study presents a sensitive determination of biogenic monoamines in rat brain striatum microdialysates using capillary high-performance liquid chromatography with the photoluminescence following electrontransfer detection technique. Separation conditions were optimized by changing the concentration of an ion-interaction agent and the percentage of an organic modifier. The high concentration of ion-interaction agent enabled the amines as a class to be separated from interfering acids, but also made the separation very long. To shorten the separation time, 10% (v/v) acetonitrile was used as the organic modifier. Eight chromatographic runs during a 3-h period were analyzed in terms of retention times, peak heights, and peak widths. Chromatograms are very reproducible, with less than 1% changes in peak height over 3 h. Typical concentration detection limits at the optimum separation conditions were less than 100 pM for metabolic acids and similar to 200 pM for monoamines. The injection volume of the sample was 500 nL. Thus, the mass detection limits were less than 50 amol for metabolic acids and similar to 100 amol for monoamines. Typical separation time was less than 10 min. To validate the technique, the separation method was applied to the observation of drug-induced changes of monoamine concentrations in rat brain microdialysis samples. Local perfusion of tetrodotoxin, a sodium channel blocker, into the striatum of an anesthetized rat decreased dopamine, 3-methoxytyramine, and serotonin concentrations in dialysates. Successive monitoring of striatal dialysates at a temporal resolution of 7.7 min showed that the injection of nomifensine transiently increased dopamine and 3-methoxytyramme concentrations in rat brain dialysate.