Direct assay of glutathione peroxidase activity using high-performance capillary electrophoresis.
J Chromatogr
; 581(1): 49-56, 1992 Oct 02.
Article
em En
| MEDLINE
| ID: mdl-1430007
ABSTRACT
A fast, sensitive and direct method has been developed for the determination of glutathione peroxidase activity (both selenium- and non-selenium-dependent) in cell-free preparations. The assay is based on the separation and quantitation of reduced and oxidized glutathione by capillary electrophoresis. The electrophoretic separation buffer was 100 mM sodium tetraborate (pH 8.2) containing 100 mM sodium dodecylsulphate. A micellar electrokinetic mechanism took place under these conditions, and a total mass recovery was observed for both peptides. The reproducibility of migration times was excellent (less than 3% variability). A linear detector response range was observed in the range 5-50 U/ml, and both the reproducibility and accuracy were satisfied. Samples out of this linear range could be analysed by either increasing the reaction time or diluting the enzyme preparation. The results obtained with the new direct capillary electrophoresis assay were compared with those derived from a reversed phase high-performance liquid chromatographic and spectrophotometric coupled assay. A very good agreement was found between the two direct assay methods in all samples. Capillary electrophoresis is a versatile technique that allows the automation of the glutathione peroxidase assay in a reproducible manner and within a relatively short time with sufficient accuracy and precision.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Eletroforese
/
Glutationa Peroxidase
Limite:
Animals
Idioma:
En
Revista:
J Chromatogr
Ano de publicação:
1992
Tipo de documento:
Article
País de afiliação:
Espanha