Rapid labeling of amino acid neurotransmitters with a fluorescent thiol in the presence of o-phthalaldehyde.
Rapid labeling of amino acid neurotransmitters with a fluorescent thiol in the presence of o-phthalaldehyde.
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DOI：
10.1002/elps.201600374
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发表时间：
2017-02
期刊：
影响因子：
2.9
通讯作者：
Gong M
中科院分区：
文献类型：
--
作者：
Maddukuri N;Zhang Q;Zhang N;Gong M
Laser-induced fluorescence (LIF) detection often requires labelling of analytes with fluorophores; and fast fluorescent derivatization is valuable for high-throughput analysis with flow-gated capillary electrophoresis (CE). Here, we report a fast fluorescein-labelling scheme for amino neurotransmitters, which were then rapidly separated and detected in flow-gated CE. This scheme was based on the reaction between primary amines and o-phthalaldehyde (OPA) in the presence of a fluorescent thiol, 2-[(5-fluoresceinyl)aminocarbonyl]ethyl mercaptan (FACE-SH). The short reaction time (<30 s) was suited for on-line mixing and derivatization that was directly coupled with flow-gated CE for rapid electrophoretic separation and sensitive LIF detection. To maintain the effective concentration of reactive FACE-SH, Tris(2-carboxyethyl)phosphine (TCEP) was added to the derivatization reagents to prevent thiol loss due to oxidation. This labelling scheme was applied to the detection of neurotransmitters by coupling in vitro microdialysis with on-line derivation and flow-gated CE. It is also anticipated that this fluorophore tagging scheme would be valuable for on-chip labelling of proteins retained on support in solid phase extraction.
