Diethylation Labeling Combined with UPLC/MS/MS for Simultaneous Determination of a Panel of Monoamine Neurotransmitters in Rat Prefrontal Cortex Microdialysates
Diethylation Labeling Combined with UPLC/MS/MS for Simultaneous Determination of a Panel of Monoamine Neurotransmitters in Rat Prefrontal Cortex Microdialysates
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DOI：
10.1021/ac801339z
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发表时间：
2008-12-01
影响因子：
7.4
通讯作者：
Lepsy, Christopher
中科院分区：
文献类型：
--
作者：
Ji, Chengjie;Li, Wenlin;Lepsy, Christopher
与开发用于同时测定大鼠脑微透析液中的生物源性单胺神经递质（如去甲肾上腺素（NE）、多巴胺（DA）、5-羟色胺（5-HT）和去甲甲肾上腺素（NM））的LC/MS/MS分析相关的主要挑战是提高检测灵敏度。在这项工作中，UPLC/MS/MS为基础的方法结合二乙基标记技术，同时测定大鼠前额叶皮层微透析液中的一组单胺。色谱运行时间为3.5 min/样品。NE、DA、5-HT/和NM的基于UPLC/MS/MS的方法（含/不含单胺二乙基标记）的检测限分别为0.005/0.4（30/2367 pM）、0.005/0.1（33/653 pM）、0.005/0.2（28/1136 pM）和0.002/0.2 ng/mL（11/1092 pM）。在UPLC/MS/MS分析期间，单胺的氨基的二乙基标记提供了相应天然单胺的20-100倍的检测灵敏度增加。这可能是由于以下原因：（1）改进的裂解模式;（2）增加的疏水性和伴随的ESI MS和MS/MS分析中的电离效率增加;（3）减少的基质干扰。该标记反应采用市售试剂，乙酰丙酮-d（4），通过还原胺化标记单胺上的胺基。在温和的反应条件下，它还具有简单、快速（类似于25分钟的反应时间）、特异性和定量性。数据还显示，从应用该测定监测药物诱导的单胺浓度的变化，在大鼠前额叶皮层微透析液样品，然后由SKF 81297，一种选择性的D，多巴胺受体激动剂，已知提高神经递质DA和NE在中枢神经系统的细胞外水平的管理。
The primary challenge associated with the development of an LC/MS/MS-based assay for simultaneous determination of biogenic monoamine neurotransmitters such as norepinephrine (NE), dopamine (DA), serotonin (5-HT), and normetanephrine (NM) in rat brain microdialysates is to improve detection sensitivity. In this work, a UPLC/MS/MS-based method combined with a diethyl labeling technique was developed for simultaneous determination of a panel of monoamines in rat prefrontal cortex microdialysates. The chromatographic run time is 3.5 min/sample. The limits of detection of the UPLC/MS/MS-based method for NE, DA, 5-HT/ and NM, with/without diethyl labeling of monoamines, are 0.005/0.4 (30/2367 pM), 0.005/0.1 (33/653 pM), 0.005/0.2 (28/1136 pM), and 0.002/0.2 ng/mL (11/1092 pM), respectively. Diethyl labeling of amino groups of monoamines affords 20-100 times increased detection sensitivity of corresponding native monoamines during the UPLC/MS/MS analysis. This could result from the following: (1) improved fragmentation patterns; (2) increased hydrophobicity and concomitantly increased ionization efficiency in ESI MS and MS/MS analysis; (3) reduced matrix interference. This labeling reaction employs a commercially available reagent, acetaldehyde-d(4), to label the amine groups on the monoamines via reductive amination. It is also simple, fast (similar to 25-min reaction time), specific, and quantitative under mild reaction conditions. Data are also presented from the application of this assay to monitor the drug-induced changes of monoamine concentrations in rat prefrontal cortex microdialysate samples followed by administration of SKF 81297, a selective D, dopamine receptor agonist known to elevate the extracellular level of the neurotransmitters DA and NE in the central nervous system.