High-performance liquid chromatography method for the determination of protein-glutathione mixed disulfide.
We have developed an efficient and sensitive method for the quantification of picomole quantities of protein-glutathione and protein-cysteine mixed disulfides in ocular lens. Performic acid oxidation releases glutathione and cysteine from the protein-mixed disulfides as glutathione sulfonic acid and cysteic acid, respectively. The sulfonic acids are derivatized with phenylisothiocyanate, and the phenylthiocarbamyl derivatives are separated by HPLC using a reversed-phase ODS column, equilibrated with 0.14 M sodium acetate, containing 0.1% triethylamine and 6% acetonitrile.
Kumari
K
K
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston 77555.
Khanna
P
P
Ansari
N H
NH
Srivastava
S K
SK
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003459:Crystallins / Q000032:analysis*; D003545:Cysteine / Q000032:analysis*; D004220:Disulfides / Q000032:analysis*; D005021:Ethylamines; D005978:Glutathione / Q000032:analysis*; D007202:Indicators and Reagents; D017879:Isothiocyanates; D007908:Lens, Crystalline / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013451:Sulfonic Acids / Q000032:analysis; D013861:Thiocyanates
D003459:Crystallins; D004220:Disulfides; D005021:Ethylamines; D007202:Indicators and Reagents; D017879:Isothiocyanates; D011506:Proteins; D013451:Sulfonic Acids; D013861:Thiocyanates; C005441:phenylisothiocyanate; D005978:Glutathione; D003545:Cysteine; C016162:triethylamine