title: A novel HPLC-electrochemical detection approach for the determination of D-penicillamine in skin specimens.
abstract: D-penicillamine is a thiol drug mainly used for Wilson's disease, rheumatoid arthritis and cystinuria. Adverse effects during normal use of the drug are frequent and may include skin lesions. To evaluate its toxic effects in clinical cases an original method based on high performance liquid chromatography coupled to amperometric detection in a specific biological matrix such as skin has been developed. The chromatographic analysis of D-penicillamine was carried out on a C18 column using a mixture of acid phosphate buffer and methanol as the mobile phase. Satisfactory sensitivity was obtained by oxidizing the molecule at +0.95 V with respect to an Ag/AgCl reference electrode. A chemical reduction of D-penicillamine-protein disulphide bonds using dithioerythritol combined with microwaves was necessary for the determination of the total amount of D-penicillamine in skin specimens. A further solid-phase extraction procedure on C18 cartridges was implemented for the sample clean-up. The whole analytical procedure was validated: high extraction yield (>91.0%) and satisfactory precision (RSD<6.8%) values were obtained. It was successfully applied to skin samples from a patient who was previously under a long-term, high-dose treatment with the drug and presented serious D-penicillamine-related dermatoses. Thus, the method seems to be suitable for the analysis of D-penicillamine in skin tissues.
authors.affiliation: Department of Pharmacy and Biotechnology (FaBiT), University of Bologna, Via Belmeloro 6, I-40126 Bologna, Italy.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Saracino
authors.forename: Cristina
authors.initials: C
authors.lastname: Cannistraci
authors.forename: Francesca
authors.initials: F
authors.lastname: Bugamelli
authors.forename: Emanuele
authors.initials: E
authors.lastname: Morganti
authors.forename: Iria
authors.initials: I
authors.lastname: Neri
authors.forename: Riccardo
authors.initials: R
authors.lastname: Balestri
authors.forename: Annalisa
authors.initials: A
authors.lastname: Patrizi
authors.forename: Maria A
authors.initials: MA
authors.lastname: Raggi
mesh_terms: D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D004563:Electrochemistry; D004566:Electrodes; D005260:Female; D006527:Hepatolenticular Degeneration / Q000150:complications / Q000188:drug therapy; D006801:Humans; D008875:Middle Aged; D010396:Penicillamine / Q000009:adverse effects / Q000032:analysis* / Q000737:chemistry; D012867:Skin / Q000378:metabolism*; D012871:Skin Diseases / Q000139:chemically induced / Q000378:metabolism*; D052616:Solid Phase Extraction
chemical_list: D004220:Disulfides; D010396:Penicillamine