Determination of D-penicillamine in serum by fluorescence derivatization and liquid column chromatography

A simple and fast method for the determination of D-penicillamine in serum is described. The analysis is based o a fluorescence derivatization of the sulfhydryl group combined with a reversed-phase liquid chromatograhic separation and fluorescence detection. Before derivatization the serum proteins are precipitated with ethanol and removed by centrifugation. As derivatizing agent 5-dimethylaminonaphthaline-1-sulfonylaziridine is used which reacts selectively with thiols under defined reaction conditions. The detection limit is in the pmol range; 50-300 ng of D-penicillamine can be determined with a relative standard deviation of 7-8%. Thus the method permits a simple determination of D-penicillamine in serum at therapeutic levels.