Glutathione measurement by high-performance liquid chromatography separation and fluorometric detection of the glutathione-orthophthalaldehyde adduct.
Glutathione reacts with orthophthalaldehyde to form a stable, highly fluorescent tricyclic derivative which is easily separated and quantitated by high-performance liquid chromatography. Separation of the glutathione adduct is achieved by isocratic elution over a reverse-phase column with 7.5% methanol/92.5% 0.15 M sodium acetate, pH 7.00. The adduct is detected fluorometrically and quantitated by integration of peak area. Detection of 0.1 to 200 pmol glutathione produces a linear response and the recovery of reduced and oxidized glutathione from rat liver homogenate, bile, and plasma is quantitative. The chemical identity of the adduct was confirmed by mass spectrometry.
University of California San Francisco, Liver Center Lab, San Francisco General Hospital 94110.
B A
BA
Neuschwander-Tetri
F J
FJ
Roll
D000818:Animals; D001646:Bile / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D005978:Glutathione / Q000032:analysis* / Q000097:blood; D007202:Indicators and Reagents; D008099:Liver / Q000032:analysis; D051381:Rats; D013050:Spectrometry, Fluorescence; D009764:o-Phthalaldehyde
D007202:Indicators and Reagents; D009764:o-Phthalaldehyde; D005978:Glutathione