Determination of S-nitrosoglutathione in erythrocytes by capillary zone electrophoresis.
A method for separation and quantification of S-nitrosoglutathione in red cell extracts by capillary electrophoresis is reported. The method is based on the direct analysis of the metaphosphoric acid erythrocyte extract containing diethylenetriaminepentaacetic acid. Optimization of the method is briefly discussed. Best results in the shortest time were obtained at 25 degrees C, using a coated capillary, 7 kV applied voltage and phosphate sodium 40 mmol/L (pH 2.2) as running buffer. Reproducibility, detection limits, and recoveries of S-nitrosoglutathione analyses were checked. The results evidenced that S-nitrosoglutathione is formed in erythrocytes treated with S-nitrosocysteine, a transnitrosating agent. Under our experimental conditions, the contemporaneous detection and quantification of reduced and oxidized glutathione present in cell extract could also be performed.
Messana
I
I
Instituto di Chimica e Chimica Clinica della Facoltà di Medicina e Chirurgia dell'Università Cattolica, Roma, Italy.
Rossetti
D V
DV
Misiti
F
F
Vincenzoni
F
F
Tellone
E
E
Giardina
B
B
Castagnola
M
M
D019075:Electrophoresis, Capillary / Q000379:methods; D004912:Erythrocytes / Q000737:chemistry*; D005978:Glutathione / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D019803:Glutathione Disulfide / Q000737:chemistry; D006801:Humans; D009603:Nitroso Compounds / Q000097:blood*; D026422:S-Nitrosoglutathione
D009603:Nitroso Compounds; D026422:S-Nitrosoglutathione; D005978:Glutathione; D019803:Glutathione Disulfide