Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
39, Number 9, September 2001, pp. 365-369
N-Hydroxysuccinimidyl-Fluorescein-O-Acetate
for Precolumn Fluorescence Derivatization of Amino Acids and Oligopeptides in
Liquid ChromatographyHong
Wang, Jin Li, Ting-Xian Yang, and Hua-Shan Zhang
Department of Chemistry, Wuhan University, Wuhan 430072, China
A new fluorescent derivatizing reagent, N-hydroxysuccinimidyl-fluorescein-O-acetate,
is used for the high-performance liquid chromatographic analysis of amino acids
and oligopeptides. This reagent has the advantages of high-detection sensitivity
in the visible region, specifically with amino groups, mild derivatization conditions,
and little interference induced. The fluorescence properties of the reagent
and its derivatives with amino acids and oligopeptides are studied. The conditions
of the derivatization are investigated in detail. In the mobile phase of methanol–water
(42:58, v/v) containing a 10mM pH 5.0 citric acid–Na2HPO4
buffer, six amino acids and oligopeptides are separated in 20 min with fluorescence
detection at excitation and emission wavelengths of 492 and 513 nm, respectively,
with the detection limits for injected standards ranging from 0.64 to 12 fmol.
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