Reagent peak-free liquid chromatography-fluorescence analysis of carboxylic acids using a fluorous scavenging-derivatization method.
Anal Bioanal Chem
; 397(6): 2409-16, 2010 Jul.
Article
em En
| MEDLINE
| ID: mdl-20509021
ABSTRACT
We developed a fluorous scavenging-derivatization method for reagent peak-free liquid chromatography (LC)-fluorescence analysis of carboxylic acids. In this method, carboxylic acids were fluorescently derivatized with 1-pyrenemethylamine in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 1-hydroxy-1H-benzotriazole. Residual excess unreacted reagent was tagged with 2-(perfluorooctyl)ethyl isocyanate and could be selectively removed by microfluorous solid-phase extraction before LC analysis. With use of this method, eight fluorescent derivatives of linear aliphatic carboxylic acids (C(1)-C(8)) can be separated within 30 min by reversed-phase LC with gradient elution. In the chromatogram obtained, the fluorous-tagged unreacted reagent peak is greatly decreased after microfluorous solid-phase extraction and does not interfere with the quantification of each acid. With use of microfluorous solid-phase extraction with 80% (v/v) aqueous methanol elution, over 99.9% of the unreacted fluorescent reagent was removed. The detection limits (signal-to-noise ratio of 3) for the carboxylic acids examined are 2.3-8.0 fmol per 10-microL injection. We also applied this method successfully to the analysis of highly polar carboxylic acids such as alpha-keto acids and tricarboxylic acid cycle metabolites.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ácidos Carboxílicos
/
Cromatografia Líquida
Idioma:
En
Revista:
Anal Bioanal Chem
Ano de publicação:
2010
Tipo de documento:
Article
País de afiliação:
Japão