Determination of penicillamine, penicillamine disulfide and penicillamine-glutathione mixed disulfide by high-performance liquid chromatography with electrochemical detection.
Methodology is described for the simultaneous determination of D-penicillamine, penicillamine disulfide and the penicillamine-glutathione mixed disulfide, as well as glutathione and glutathione disulfide, in human plasma, erythrocytes and urine. The various thiols and disulfides are separated by reversed-phase ion-pairing liquid chromatography with detection by an electrochemical detector with dual gold/mercury amalgam electrodes in series. The thiols are detected at the downstream electrode; the disulfides are reduced at the upstream electrode and then detected as the thiols at the downstream electrode. Detection limits (at a signal-to-noise ratio of 2.0) are in the picomole range for 20 microliters of injected solution for all compounds except penicillamine disulfide, which has a detection limit of 600 pmol in 20 microliters. A convenient method is described for preparation of the penicillamine-glutathione mixed disulfide by thiol/disulfide exchange with standardization of the solution by 1H NMR spectroscopy.
Yamashita
G T
GT
Department of Chemistry, University of Alberta, Edmonton, Canada.
Rabenstein
D L
DL
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D004220:Disulfides / Q000032:analysis*; D004563:Electrochemistry; D004912:Erythrocytes / Q000032:analysis; D005978:Glutathione / Q000032:analysis / Q000031:analogs & derivatives; D019803:Glutathione Disulfide; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010396:Penicillamine / Q000032:analysis* / Q000031:analogs & derivatives
D004220:Disulfides; C036069:penicillamine disulfide; C061524:penicillamine-glutathione mixed disulfide; D005978:Glutathione; D010396:Penicillamine; D019803:Glutathione Disulfide