Abstract
N-Hydroxysuccinimidyl-3-indolylacetate (SIIA) is a new fluorescent derivatizing reagent with an indole ring and ¶an N-hydroxysuccinimide ester functionality. It can react with catecholamines under mild conditions to form corresponding amides, which have strong fluorescence at λex/λem = 301 nm/¶365 nm. This paper covers the RP-HPLC separation and fluorescent determination of derivatized catecholamines with SIIA. In a mobile phase of methanol-water (36/64, v/v) containing H3cit-Na2HPO4 buffer (pH = 4.00, 10 mmol/L), the derivatives of norepinephrine (NE), epinephrine (E) and dopamine (DA) were eluted within 15 min on a C18 column. The detection limits were 0.043, 0.13 and 0.18 pmol, respectively, when the ratio of signal to noise (S/N) was 3. The excessive reagent is rapidly hydrolyzed to 3-indolylacetic acid (IA) that can be easily separated from derivatives.
Similar content being viewed by others
Author information
Authors and Affiliations
Additional information
Received: 16 April 1999 / Revised: 13 July 1999 / /Accepted: 15 July 1999
Rights and permissions
About this article
Cite this article
Wang, H., Jin, H. & Zhang, Hs. Determination of catecholamines as their N-hydroxy-succinimidyl-3-indolylacetate derivatives by pre-column derivatization HPLC separation and fluorescent detection. Fresenius J Anal Chem 365, 682–684 (1999). https://doi.org/10.1007/s002160051545
Issue Date:
DOI: https://doi.org/10.1007/s002160051545