Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC-MS/MS to simultaneously quantify dopamine, serotonin and -aminobutyric acid in four rat brain regions
Development, validation and comparison of surrogate matrix and surrogate analyte approaches with UHPLC-MS/MS to simultaneously quantify dopamine, serotonin and -aminobutyric acid in four rat brain regions
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使用 UHPLC-MS/MS 开发、验证和比较替代基质和替代分析物方法，以同时定量四个大鼠大脑区域中的多巴胺、血清素和 β-氨基丁酸
DOI：
10.1002/bmc.4276
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发表时间：
2018-09-01
影响因子：
1.8
通讯作者：
Liu, Wan-hui
中科院分区：
文献类型：
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作者：
Liu, Rong-xia;Xian, You-yan;Liu, Wan-hui
内源性神经递质作为生物标志物，在神经精神疾病的发生发展过程中起着重要作用，不同脑区神经递质水平的检测有助于神经系统疾病的诊断和治疗。由于缺乏内源性神经递质的空白基质，替代基质和替代分析物方法已被用于测定神经递质来解决这个问题。在这项研究中，我们利用了UHPLC-MS/MS的高准确度，精密度和通量，并开发了基于这两种方法的新方法。这两种方法满足FDA和EMA的验证标准后，适当的平行评估，他们被用来进一步定量的三种内源性神经递质，包括多巴胺（DA），5-羟色胺（5-HT）和-氨基丁酸（GABA）在大鼠大脑的四个区域（皮层，纹状体，下丘脑和海马），分别代表的儿茶酚胺，吲哚胺和氨基酸，。对10只相同的大鼠进行比较，两种方法对DA、5-HT、GABA的影响无显著性差异（P > 0.05）。纹状体和下丘脑的DA和GABA含量最高，纹状体和海马的5-HT含量最高，约为其他区域的2倍。这是第一个研究，比较这两种方法在测定大鼠脑内源性神经递质含量，和替代矩阵的方法被证明是简单，快速，可靠，考虑成本，矩阵的相似性，和实用性。
As biomarkers, endogenous neurotransmitters play critical roles in the process of neuropsychiatric diseases, and neurotransmitter levels in different brain regions can contribute to neurological disease diagnosis and treatment. Due to the lack of a blank matrix for endogenous neurotransmitters, surrogate-matrix and surrogate-analyte approaches have been used for the determination of neurotransmitters to solve this problem. In this study, we capitalised on the high accuracy, precision, and throughput of UHPLC-MS/MS and developed new methods based on the two approaches. Both approaches satisfied FDA and EMA validation criterias after an appropriate parallelism assessment, and they were used to further quantify the three endogenous neurotransmitters, including dopamine (DA), serotonin (5-HT) and -aminobutyric acid (GABA) in rat brain four regions (cortex, striatum, hypothalamus and hippocampus) which represent the catecholamines, indolamines, and amino acids, respectively. Comparison of the results in the same rats (n=10) showed there was no significant difference in DA, 5-HT, or GABA levels between the two approaches (P > 0.05). The concentrations of DA and GABA were highest in striatum and hypothalamus, respectively, and the levels of 5-HT were paralleled in striatum and hippocampus almost 2-fold higher than other regions. This is the first study to compare these two approaches in the determination of endogenous neurotransmitter content in the rat brain, and the surrogate-matrix approach proved to be simple, rapid, and reliable, considering cost, matrix similarity, and practicality.