Department Biomedical Sciences and Centre of Excellence for Biotechnology Development and Biodiversity Research, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy.
Anal Bioanal Chem. 2010 Nov;398(5):1973-8. doi: 10.1007/s00216-010-4134-5. Epub 2010 Aug 28.
The role of neurotransmitter amino acids (NAAs) in the functioning of the nervous system has been the focus of increasingly intense research over the past several years. Among the various amino acids that have important roles as neurotransmitters, there are alanine (Ala), glutamic acid (Glu), aspartic acid (Asp), serine (Ser), taurine (Tau) and glycine (Gly). NAAs are present in plasma, cells and--at trace levels--in all biological fluids, but complex components in biological matrices make it difficult to determine them in biological samples. We describe a new capillary electrophoresis (CE) method with laser-induced fluorescence detection by which analytes are resolved in less than 12 minutes in a 18 mmol/L phosphate run buffer at pH 11.6. The use of elevated temperatures during sample derivatization leads to a drastic reduction in the reaction time, down to 20 min, compared to the 6-14 h usually described for reactions between FITC and amino acids at room temperature. In order to demonstrate its wide range of applications, the method was applied to the analysis of NAA in human plasma and in other sample types, such as red blood cells, urine, cultured cells, cerebrospinal fluid, saliva and vitreous humor, thus avoiding the typical limitations of other methods, which are normally suitable for use with only one or two matrix types.
在过去的几年里，神经递质氨基酸（NAAs）在神经系统中的作用一直是研究的焦点。在作为神经递质发挥重要作用的各种氨基酸中，有丙氨酸（Ala）、谷氨酸（Glu）、天冬氨酸（Asp）、丝氨酸（Ser）、牛磺酸（Tau）和甘氨酸（Gly）。NAAs 存在于血浆、细胞中，以及所有生物体液中的痕量水平，但生物基质中的复杂成分使得难以在生物样本中确定它们。我们描述了一种新的毛细管电泳（CE）方法，采用激光诱导荧光检测，在 18mmol/L 磷酸盐运行缓冲液（pH11.6）中，不到 12 分钟即可分离分析物。在样品衍生化过程中使用升高的温度会导致反应时间大大缩短，与室温下 FITC 与氨基酸之间通常描述的 6-14 小时相比，反应时间缩短至 20 分钟。为了展示其广泛的应用，该方法应用于人血浆和其他样本类型（如红细胞、尿液、培养细胞、脑脊液、唾液和玻璃体）中 NAA 的分析，从而避免了其他方法通常只适用于一种或两种基质类型的典型限制。
