SIMULTANEOUS MEASUREMENT OF MONOAMINES, METABOLITES AND AMINO-ACIDS IN BRAIN-TISSUE AND MICRODIALYSIS PERFUSATES
SIMULTANEOUS MEASUREMENT OF MONOAMINES, METABOLITES AND AMINO-ACIDS IN BRAIN-TISSUE AND MICRODIALYSIS PERFUSATES
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DOI：
10.1016/0378-4347(93)80311-q
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发表时间：
1993-05-05
期刊：
影响因子：
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通讯作者：
ACWORTH, IN
中科院分区：
文献类型：
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作者：
GAMACHE, P;RYAN, E;ACWORTH, IN
建立了一种同时测定单胺类化合物及其代谢产物和邻苯二甲醛衍生氨基酸的高效液相色谱-库仑阵列电化学检测方法。该方法已被用于检查纹状体组织匀浆和纹状体微透析灌注液中的代谢物水平。通过等度洗脱和电化学检测，在8个库仑流通石墨电极（0至490 mV; 70 mV增量）的串联电极阵列上，对等份样品进行单胺和代谢物的初步分析。剩余样品用OPA-β-巯基乙醇进行柱前衍生化，柱切换后，在第二个等度系统上分析氨基酸，在四个电极上进行电化学检测。然后基于它们的保留时间以及跨阵列的电化学行为来鉴定金属离子。分析、衍生化程序、色谱柱切换、数据简化和峰鉴别完全自动化。纹状体组织匀浆的检测限约为1.38纳克/克湿重的单胺和8.25纳克/克湿重的氨基酸。纹状体灌流液的检测限对于单胺为约2.5 pg/20-穆尔样品，对于氨基酸为15 pg/20-穆尔样品，分析在25分钟内完成，使得其对于微透析样品是理想的。
A high-performance liquid chromatographic method with coulometric array electrochemical detection is described for the simultaneous analysis of monoamines, their metabolites and o-phthalaldehyde (OPA)-derivatized amino acids. This method has been used to examine metabolite levels in both striatal tissue homogenates and striatal microdialysis perfusates. An aliquot of sample was initially analyzed for monoamines and metabolites by isocratic elution and electrochemical detection on a serial electrode array of eight coulometric flow-through graphite electrodes (0 to 490 mV; 70-mV increment). The remaining sample was derivatized pre-column with OPA-beta-mercaptoethanol and after column switching was analyzed for amino acids on a second isocratic system with electrochemical detection on four electrodes. Metabolites were then identified based on their retention time as well as electrochemical behavior across the arrays. The analysis, derivatization procedure, column switching, data reduction and peak identification were fully automated. The limit of detection for striatal tissue homogenates was approximately 1.38 ng/g wet weight for the monoamines and 8.25 ng/g wet weight for amino acids. The limit of detection for striatal perfusates was approximately 2.5 pg per 20-mul sample for the monoamines and 15 pg per 20-mul sample for the amino acids with analysis completed within 25 min making it ideal for microdialysis samples.