Picomole analysis of glutathione, glutathione disulfide, glutathione S-sulfonate, and cysteine S-sulfonate by high-performance liquid chromatography.
A method for simultaneous detection of picomole quantities of glutathione (GSH), glutathione disulfide (GSSG), glutathione S-sulfonate (GSSO3H), and cysteine S-sulfonate (CYSSO3H) by high-performance liquid chromatography has been developed. Compounds are separated by anion-exchange chromatography using a citric acid buffer system, and then derivatized postcolumn using o-phthalaldehyde with 2-mercaptoethanol, heated to 70 degrees C, and detected by fluorescence. The compounds elute with retention times of 12.5 min for GSH, 27.5 min for CYSSO3H, 29.8 min for GSSG, and 33.0 minutes for GSSO3H, with detection limits of 10, 200, 10, and 50 pmol, respectively. Recoveries are 103% for GSH, 102% for GSSG, 100% for CYSSO3H, and 96% for GSSO3H. Determination of target compounds in cells is described.
D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003545:Cysteine / Q000031:analogs & derivatives* / Q000032:analysis; D005978:Glutathione / Q000031:analogs & derivatives* / Q000032:analysis*; D019803:Glutathione Disulfide; D006801:Humans; D008168:Lung / Q000032:analysis; D008175:Lung Neoplasms; D008832:Microchemistry; D051381:Rats; D012710:Serum Albumin, Bovine
C040136:glutathione sulfonate; D012710:Serum Albumin, Bovine; C011119:S-sulphocysteine; D005978:Glutathione; D003545:Cysteine; D019803:Glutathione Disulfide