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N-羟基琥珀酰亚胺-3-吲哚乙酸酯柱前衍生HPLC分离荧光检测氨基酸
DETERMINATION OF AMINO ACIDS AS THEIR N HYDROXY SUCCINIMIDYL 3 INDOLYLACETATE DERIVATIVES BY PRE COLUMN DERIVATIZATION HPLC SEPARATION AND FLUORESCENCE DETECTION
【摘要】 N-羟基琥珀酰亚胺-3-吲哚乙酸酯(SIIA)是一种含有新型反应活性基团的荧光衍生试剂,可在温和的条件下与氨基化合物进行衍生反应.以SIA为柱前衍生试剂,研究了其氨基酸衍生物的高效液相色谱行为和分离条件,在含适宜pH值柠檬酸(H3cit)-Na2HPO4缓冲溶液的甲醇-水流动相中,获得了常见氨基酸的有效分离,检出限达亚pmol级.
【Abstract】 N hydroxy succinimidyl 3 indolylacetate (SIIA) is a new fluorescence derivatization reagent with an indole ring and a N hydroxy succinimide ester reactive function. It can react with amino compounds under mild conditions and yields corresponding amides, which have strong fluorescence at λ ex / λ em =278 nm/355 nm. This paper covers the reversed phase HPLC separation and determination of sixteen derivatized amino acids with SIIA by a mobile phase of methanol water containing H 3cit Na 2HPO 4 buffer. The excess reagent will be rapidly hydrolyzed to give 3 indolylacetic acid which can be easily separated from derivatives, so it does not interfere with the analysis. The study showed that SIIA can be used as a derivatization reagent for the analysis of amino compounds by reversed phase HPLC and fluorescence detection.
【Key words】 N hydroxy succinimidyl 3 indolylacetate; fluorescence derivatization reagent; aminoacid; high performance liquid chromatography (HPLC);
- 【文献出处】 武汉大学学报(自然科学版) ,WUHAN UNIVERSITY JOURNAL(NATURAL SCIENCE EDITION) , 编辑部邮箱 ,1998年02期
- 【分类号】O629.71,O657.32
- 【被引频次】4
- 【下载频次】178