中央民族大学学报:自然科学版
中央民族大學學報:自然科學版
중앙민족대학학보:자연과학판
Journal of The Central University for Nationalities(Natural Sciences Edition)
2012年
2期
19-22
,共4页
高效液相色谱%柱前衍生%荧光检测%儿茶酚胺
高效液相色譜%柱前衍生%熒光檢測%兒茶酚胺
고효액상색보%주전연생%형광검측%인다분알
high performance liquid chromatography%pre-column derivatization%fluorescence detection%catecholamines
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.
利用新型熒光試劑1,3,5,7-四甲基-8-苯基-(4'-O-(N-琥珀酰亞胺乙痠酯))-二氟化硼-二吡咯甲烷(TMPAB-OSu)為柱前衍生化試劑,在氰基色譜柱上通過梯度洗脫對三種兒茶酚胺神經遞質進行瞭分離和檢測.以乙腈-水為溶劑,pH7.5的硼痠-硼砂緩遲溶液中在30℃條件下衍生反應5 min後穫得穩定的熒光衍生產物.激髮波長和髮射波長分彆為497 nm和509 nm,實現瞭兒茶酚胺柱前衍生檢測,方法的重現性好、靈敏度高.兒茶酚胺的線性相關繫數大于0.9970,檢齣限為5 nmol/L.
이용신형형광시제1,3,5,7-사갑기-8-분기-(4'-O-(N-호박선아알을산지))-이불화붕-이필각갑완(TMPAB-OSu)위주전연생화시제,재청기색보주상통과제도세탈대삼충인다분알신경체질진행료분리화검측.이을정-수위용제,pH7.5적붕산-붕사완충용액중재30℃조건하연생반응5 min후획득은정적형광연생산물.격발파장화발사파장분별위497 nm화509 nm,실현료인다분알주전연생검측,방법적중현성호、령민도고.인다분알적선성상관계수대우0.9970,검출한위5 nmol/L.
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 rim. 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.