毛细管电泳—激光诱导荧光法分离四种神经递质类氨基酸的条件优化

Optimization of Seperating 4 Neurotransmitter Amino Acids by Capillary Electrophoresis with Laser-induced Fluorescence

目的探讨毛细管电泳分离神经递质类氨基酸的影响因素。方法采用毛细管电泳法分离4种神经递质类氨基酸的荧光素异硫氰酸酯(FITC-AAs)衍生物,分析电泳缓冲液、表面活性剂以及衍生条件对分离的影响。结果电泳缓冲液的组成、浓度、pH值、表面活性剂以及衍生条件对4种FITC-AA衍生物分离有很大影响,在以75 mm ol/L,pH 9.42硼酸盐含100 mm ol/LSDS为电泳缓冲液,50 mmol/LpH 9.8硼酸盐为衍生缓冲液,4种FITC-AA衍生物20 min内得到很好分离。结论通过条件优化,毛细管电泳-激光诱导荧光技术分离神经递质类氨基酸的分离效果能得到很大提高。

Objective To investigate main influential factors for determination of neurotransmitter amino acids by capillary electrophoresis. Methods 4 neurotransmitter amino acids derivatized with FITC were determined by capillary electrophoresis. Buffer concentrations,pH,surfactants and derivatization conditions were studied and optimized for fast and efficient separation. Results The running buffer concentrations,pH,surfactants and derivatization conditions could be improved for the separating efficiency. 4 neurotransmitter amino acids derivatized with FITC were separated on base line in the optimum condition of 75 mmol/L boric acid （pH 9.42） contanining 100 mmol/L SDS acting as running buffer and 50 mmol/L boric acid （pH 9.80） as derivatization buffer in 20 min. Conclusion Through optimization,the separating efficiency of capillary electrophoresis with laser--induced fluorescence detecting 4 neurotransmitter amino acids can be remarkably improved.