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抑郁大鼠脑内七种氨基神经递质水平异常及高效液相色谱-荧光检测法测定

Abnormal levels of seven amino neurotransmitters in depressed rat brain and determination by HPLC-FLD.

作者信息

Cui Ting, Qiu Hong-Mei, Huang Dan, Zhou Qi-Xin, Fu Xiao-Yan, Li Hai-Yan, Jiang Xin-Hui

机构信息

School of Pharmacy, Chongqing Medical University, Chongqing, China.

出版信息

Biomed Chromatogr. 2017 Aug;31(8). doi: 10.1002/bmc.3937. Epub 2017 Mar 2.

Abstract

The determination of amino acids with actions like neurotransmitters or modulators has been increasingly important for diagnosis in many neuropsychiatric diseases. A rapid and simple high-performance liquid chromatography with fluorescence detection method was developed for simultaneous determination of seven amino acids: aspartate (Asp), glutamate (Glu), serine (Ser), glutamine (Gln), glycine (Gly), taurine (Tau) and γ-aminobutyric` acid (GABA). Homoserine was used as an internal standard. The analysis was performed on a BDS column with methanol and 50 mm sodium acetate solution (pH 6.5) using a simple gradient elution. Several parameters of the developed method were validated including linearity, accuracy, precision, extraction recovery and stability, which were within the acceptable range. The method was successfully applied to determination of real samples: hippocampus and cortex in depressed rats exposed to chronically unpredictable stress in order to study if there existed differences in the seven amino acids levels between depressed rats and control. The results showed that Asp, Gly, Tau and GABA significantly decreased with increasing Gln in the hippocampus of depressed rats, compared with that of the control group, among which obviously lower level of Asp and higher level of Gln in cortex were observed. The analytical method and the results could be useful for clinical diagnosis and further insight into pathophysiological mechanism of depression.

摘要

对具有神经递质或调质作用的氨基酸进行测定,在许多神经精神疾病的诊断中变得越来越重要。本文建立了一种快速简便的高效液相色谱荧光检测法,用于同时测定七种氨基酸:天冬氨酸(Asp)、谷氨酸(Glu)、丝氨酸(Ser)、谷氨酰胺(Gln)、甘氨酸(Gly)、牛磺酸(Tau)和γ-氨基丁酸(GABA)。以高丝氨酸作为内标。采用简单梯度洗脱,在BDS柱上以甲醇和50 mM醋酸钠溶液(pH 6.5)进行分析。对所建立方法的几个参数进行了验证,包括线性、准确度、精密度、提取回收率和稳定性,均在可接受范围内。该方法成功应用于实际样品的测定:对慢性不可预测应激所致抑郁大鼠的海马和皮层进行检测,以研究抑郁大鼠与对照组之间这七种氨基酸水平是否存在差异。结果显示,与对照组相比,抑郁大鼠海马中Asp、Gly、Tau和GABA水平随Gln升高而显著降低,其中皮层中Asp水平明显较低,Gln水平较高。该分析方法及结果有助于临床诊断,并进一步深入了解抑郁症的病理生理机制。

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