Premium
Direct determination of glutathione in human blood by micellar electrokinetic chromatography: Simultaneous determination of lipoamide and lipoic acid
Author(s) -
Pañak Karina C.,
Ruiz Oscar A.,
Giorgieri Sergio A.,
Díaz Luis E.
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150171021
Subject(s) - lipoic acid , chromatography , chemistry , micellar electrokinetic chromatography , glutathione , detection limit , sodium dodecyl sulfate , tris , derivatization , electrokinetic phenomena , centrifugation , high performance liquid chromatography , biochemistry , antioxidant , enzyme
A reproducible, rapid procedure for the determination of glutathione in human blood by micellar electrokinetic chromatography has been developed. Whole blood samples were deproteinized with 5% w/v sulfosalicylic acid (final concentration). After centrifugation, the supernatant was directly injected for analysis, without further derivatization. Separations were performed by using an uncoated capillary of 30 cm effective length and 50 μm internal diameter (ID), 50 m M Tris‐HCl, 30 m M sodium dodecyl sulfate (SDS), pH 7.00, as running buffer, and 10–20 kV. On‐line detection was carried out at 214 nm and a detection limit in the range of femtomoles was achieved. Under the same experimental conditions, we resolved a mixed standard solution containing glutathione in its oxidized and reduced forms, lipoamide and α‐lipoic acid. The corresponding migration times were reproducible. The present method allows rapid determination of these compounds, which play a critical role in oxidative stress, in cellular defense against injurious agents and whose levels are related to the toxicology and metabolism of several toxins and drugs, such as antineoplastic agents.