Simultaneous high-performance capillary electrophoretic determination of reduced and oxidized glutathione in red blood cells in the femtomole range.
This paper describes a high-performance capillary electrophoretic (HPCE) method which allows a quick, simultaneous and quantitative determination of reduced (GSH) and oxidized (GSSG) glutathione in mammalian red blood cells using a Supelco-bonded hydrophilic phase capillary CElect-P150. The extraction procedure of GSH and GSSG from erythrocytes using Microcon-10 membranes is very simple and allows a correct evaluation of these compounds present in the red blood cells. Furthermore, the HPCE method does not require removal of the excess N-ethylmaleimide used to block the glutathione in its reduced state, making the simultaneous evaluation of GSH and GSSG possible in a very short time (ca. 4 min), with a sensitivity at femtomole level.
Piccoli
G
G
Istituto di Chimica Biologica Giorgio Fornaini, Università di Urbino, Italy.
Fiorani
M
M
Biagiarelli
B
B
Palma
F
F
Potenza
L
L
Amicucci
A
A
Stocchi
V
V
D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002197:Capillary Action; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D004912:Erythrocytes / Q000737:chemistry*; D005033:Ethylmaleimide / Q000494:pharmacology; D005978:Glutathione / Q000097:blood*; D007501:Iron / Q000494:pharmacology; D008832:Microchemistry; D010084:Oxidation-Reduction; D011817:Rabbits; D012680:Sensitivity and Specificity
D007501:Iron; D005978:Glutathione; D005033:Ethylmaleimide; D001205:Ascorbic Acid