SEPARATION AND DETERMINATION OF FLUORESCEIN ISOTHIOCYANATE-LABELED AMINO-ACIDS BY CAPILLARY-ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCEDETECTION

Citation
K. Takizawa et H. Nakamura, SEPARATION AND DETERMINATION OF FLUORESCEIN ISOTHIOCYANATE-LABELED AMINO-ACIDS BY CAPILLARY-ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCEDETECTION, Analytical sciences, 14(5), 1998, pp. 925-928
Citations number
17
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry Analytical
Journal title
ISSN journal
0910-6340
Volume
14
Issue
5
Year of publication
1998
Pages
925 - 928
Database
ISI
SICI code
0910-6340(1998)14:5<925:SADOFI>2.0.ZU;2-W
Abstract
A method for the separation and determination of amino acids by capill ary electrophoresis with laser-induced fluorescence detection has been developed. At first, the condition for the derivatization of amino ac ids with fluorescein isothiocyanate (FITC) was studied; it was proved to be optimal at 50 degrees C for 5 h. Next, the separation of 20 FITC -amino acids was attempted by using an unmodified fused silica capilla ry. Actually, 18 kinds of FITC-amino acids were separated with 96 mM s odium berate buffer (pH 9.5) containing either 25% acetone (separation system 1) or 10% methanol (separation system 2). The resolution of 19 FITC-amino acids was attained with 80 mM sodium berate buffer (pH 9.2 ) containing 45 mM alpha-cyclodextrin (separation system 3). Finally, we succeeded in separating 20 FITC-amino acids by combining two migrat ion systems (2 and 3). The FITC-amino acids were detected by argon-ion laser-induced fluorescence (Ex 488 nm, Em 520 nm). The detection limi ts of the amino acids obtained with separation systems 1 and 2 were 0. 052-1.01 fmol.