Determination of glutathione, cysteine and N-acetylcysteine in rabbit eye tissues using high-performance liquid chromatography and post-column derivatization with 5,5'-dithiobis(2-nitrobenzoic acid).
A high-performance liquid chromatographic method to determine glutathione, cysteine and N-acetylcysteine in rabbit retina, vitreous and lens has been developed. The thiols are separated using a 25 x 0.46-cm octadecylsilane column with 0.5 M phosphate buffer, pH 3, as mobile phase. The detection, at 412 nm, involves a post-column derivatization with 5,5-dithiobis(2-nitrobenzoic acid) in presence of cationic micelles of hexadecyltrimethylammonium bromide that enhances the sensitivity. The detection limits are 0.21, 0.92 and 0.61 mumol/g wet sample for glutathione, cysteine and N-acetylcysteine, respectively.
Nozal
M J
MJ
Department of Analytical Chemistry, Faculty of Sciences, University of Valladolid, Spain.
Bernal
J L
JL
Toribio
L
L
Marinero
P
P
Moral
O
O
Manzanas
L
L
Rodriguez
E
E
D000111:Acetylcysteine / Q000008:administration & dosage / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003545:Cysteine / Q000032:analysis* / Q000737:chemistry; D004228:Dithionitrobenzoic Acid / Q000737:chemistry*; D005123:Eye / Q000737:chemistry*; D005978:Glutathione / Q000032:analysis* / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007908:Lens, Crystalline / Q000737:chemistry; D011817:Rabbits; D012016:Reference Values; D012160:Retina / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013439:Sulfhydryl Reagents / Q000737:chemistry*; D014821:Vitrectomy; D014822:Vitreous Body / Q000737:chemistry
D013439:Sulfhydryl Reagents; D004228:Dithionitrobenzoic Acid; D005978:Glutathione; D003545:Cysteine; D000111:Acetylcysteine