4-Fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) has been characterized as a fluorogenic labeling reagent for high speed, online microdialysis-capillary electrophoresis (MD-CE) assays for amino acid neurotransmitters. NBD-F labeled amines are efficiently excited using 488 nm light allowing more common lasers to be used for excitation than has been previously possible. Amino acids were sampled using microdialysis, derivatized using NBD-F in an online reaction, and directly coupled to a high-speed CE separation with laser induced fluorescence (LIF) detection. Separation conditions were optimized allowing baseline separation of 16 amino acids, including glutamate, GABA, glycine, taurine, and D-serine, in 21.5 s. Parameters of the assay, including reaction temperature (80 degrees C), reaction time (5 min), NBD-F concentration (20 mM), and laser power (20 mW), were optimized. Limits of detection for glutamate and GABA were improved over previous assays by 4-fold and 25-fold, respectively. In vivo measurements were made in the rat striatum to demonstrate the effectiveness of the assay. Changes in in vivo concentrations as small as 8-9% could be observed for glycine, d-serine, and l-serine with statistical confidence. Changes in neurotransmitter concentrations induced by stimulation with high-K+ artificial cerebral spinal fluid (aCSF) were observed.

中文翻译：

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4-氟-7-硝基-2,1,3-苯并恶二唑是一种荧光标记试剂，用于使用在线微透析-毛细管电泳对氨基酸神经递质进行体内分析。

4-氟-7-硝基-2,1,3-苯并恶二唑（NBD-F）已被表征为一种荧光标记试剂，用于氨基酸神经递质的高速在线微透析-毛细管电泳（MD-CE）分析。使用488 nm光可有效激发NBD-F标记的胺，从而使比以前更常见的激光激发成为可能。使用微透析对氨基酸进行采样，在在线反应中使用NBD-F衍生化氨基酸，然后通过激光诱导荧光（LIF）检测直接与高速CE分离耦合。优化了分离条件，可以在21.5 s内基线分离16个氨基酸，包括谷氨酸，GABA，甘氨酸，牛磺酸和D-丝氨酸。测定参数，包括反应温度（80摄氏度），反应时间（5分钟），NBD-F浓度（20 mM），和激光功率（20毫瓦）进行了优化。谷氨酸和GABA的检出限分别比以前的测定法提高了4倍和25倍。在大鼠纹状体中进行了体内测量，以证明该测定的有效性。甘氨酸，d-丝氨酸和l-丝氨酸的体内浓度变化小至8-9％，具有统计上的置信度。观察到由高K +人工脑脊髓液（aCSF）刺激引起的神经递质浓度的变化。和l-丝氨酸具有统计上的信心。观察到由高K +人工脑脊髓液（aCSF）刺激引起的神经递质浓度的变化。和l-丝氨酸具有统计上的信心。观察到由高K +人工脑脊髓液（aCSF）刺激引起的神经递质浓度的变化。