Analysis of D-penicillamine in plasma by fluorescence derivatisation with N-[p-(2-benzoxazolyl)-phenyl] maleimide and high-performance liquid chromatography.
A high-performance liquid chromatographic assay for penicillamine in plasma is described. The method is based on the derivatisation of penicillamine in acidified protein-free plasma supernatants with the sulphydryl-specific reagent N-[p-(2-benzoxazolyl)-phenyl] maleimide (BOPM) to give a stable fluorescent product. After separation of the penicillamine-BOPM derivative by reversed-phase high-performance liquid chromatography, fluorescence detection enables the quantitation of plasma penicillamine concentrations in the range 0.25-500 mumol/l. The method is selective and reproducible, and since chromatography time is less than 7 min the method is readily applicable to the analysis of the large number of samples associated with pharmacokinetic studies.
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008301:Maleimides; D008832:Microchemistry; D010396:Penicillamine / Q000097:blood*; D013050:Spectrometry, Fluorescence; D013439:Sulfhydryl Reagents
D008301:Maleimides; D013439:Sulfhydryl Reagents; C038669:N-(4-(2-benzoxazolyl)phenyl)maleimide; D010396:Penicillamine