新型荧光试剂用于儿茶酚胺类神经递质的高效液相色谱检测

利用新型荧光试剂1,3,5,7-四甲基-8-苯基-(4'-O-(N-琥珀酰亚胺乙酸酯))-二氟化硼-二吡咯甲烷(TMPAB-OSu)为柱前衍生化试剂,在氰基色谱柱上通过梯度洗脱对三种儿茶酚胺神经递质进行了分离和检测.以乙腈-水为溶剂,pH7.5的硼酸-硼砂缓冲溶液中在30℃条件下衍生反应5 min后获得稳定的荧光衍生产物.激发波长和发射波长分别为497 nm和509 nm,实现了儿茶酚胺柱前衍生检测,方法的重现性好、灵敏度高.儿茶酚胺的线性相关系数大于0.9970,检出限为5 nmol/L.

Determination of Catecholamines Neurotransmitter Using Novel Pre-Column Derivatization Reagent with High Performance Liquid Chromatography

A simple and highly sensitive method based on the derivatization of catecholamines with 8-phenyl-(4-oxy-acetic acid N-hydroxysuccinimide ester)-4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene(TMPAB-OSu) as the novel pre-column derivatization reagent followed by high performance liquid chromatography with fluorescence detection was developed.Derivatization was obtained by the reaction of catecholamines with TMPAB-OSu at 30℃for 5 min in the presence of boric acid-borax buffer(pH 7.5) in acetonitrile solvent.Derivatives were sufficiently stable to be efficiently analyzed by high performance chromatography without pretreatment.On a cyan column,three catecholamines neurotransmitter were separated under gradient elution with an excitation maximum at λex 497 nm and an emission maximum at λem 509 nm.The precise determining catecholamines were obtained by post-column detection.The established method exhibited excellent reproducibility and sensitivity.Linear responses were observed with coefficients over 0.9970.The detection limits(at signal-to-noise ratio of 3:1) was 5nmol/L.