Artículo
Rapid and sensitive gradient liquid chromatography method for the quantitation of 2-hydroxyethidium ion from neutrophils
Lebed, Pablo Jorge
; Osorio Grisales, Jaiver
; Keunchkarian, Sonia
; Gotta, Javier
; Giambelluca, Miriam Soledad
; Castells, Cecilia Beatriz Marta
Fecha de publicación:
09/2011
Editorial:
RSC Publishing
Revista:
Analytical Methods
ISSN:
1759-9660
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
We developed an improved reversed-phase high-performance liquid chromatography(RPLC) assay for the rapid separation and quantitation of the 2-hydroxyethidium ion and achieved a high resolution between the ethidium and 2-hydroxyethidium ions, the latter being the specific product of the reaction between hydroethidine and the superoxide radical within a practicable analysis time. The RPLC-fluorescence method can reliably detect 2-hydroxyethidium-ion oncentrations down to 0.12 µM (or 1.2 pmol) and the signal is linear with concentration beyond 50 µM. An application of the method to neutrophil samples demonstrated that intracellular quantification of 2-hydroxyethidium was reproducible, as evidenced by low values of the relative standard deviations. The resulting analytical method combines a rapid separation of the relevant peaks with the degree of sensitivity required for use in routine biological analyses.
Palabras clave:
HPLC
,
Superoxide radical
,
2-hydroxyethidium
Archivos asociados
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Identificadores
Colecciones
Articulos(CCT - LA PLATA)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - LA PLATA
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - LA PLATA
Articulos(CIC)
Articulos de CENTRO DE INVEST.CARDIOVASCULARES (I)
Articulos de CENTRO DE INVEST.CARDIOVASCULARES (I)
Articulos(CIDEPINT)
Articulos de CENTRO DE INV EN TECNOLOGIA DE PINTURAS (I)
Articulos de CENTRO DE INV EN TECNOLOGIA DE PINTURAS (I)
Citación
Lebed, Pablo Jorge; Osorio Grisales, Jaiver; Keunchkarian, Sonia; Gotta, Javier; Giambelluca, Miriam Soledad; et al.; Rapid and sensitive gradient liquid chromatography method for the quantitation of 2-hydroxyethidium ion from neutrophils; RSC Publishing; Analytical Methods; 3; 9-2011; 593-598
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