Chemiluminescence detection coupled to liquid chromatography for the determination of penicillamine in human urine

An on-line liquid chromatography (LC) chemiluminescence detection system is proposed for the determination of penicillamine in human urine. Ion-pair reversed-phase LC using octane sulphonic acid as ion-pairing reagent allows the separation of penicillamine from the urine matrix. The chemiluminescence reaction of penicillamine with Ce(IV) in sulphuric acid sensitized by quinine is suggested for sensitive penicillamine detection in a post-column mixing set-up. The chemiluminescence response was linearly related to the concentration of penicillamine in the range 2.0-2000 mu mol l(-1) (r = 0.9994, peak height, and 0.9997 peak area, n = 10) with a detection limit (S/N = 3) of 1.0 mu mol l(-1) (100 pmol per injection). Relative standard deviations were less than 5% (n = 10) for 10-50 mu mol l(-1). The excretion rates of penicillamine in human urine of three volunteers after oral penicillamine administration were determined in the present study.