Determination of 8-methoxypsoralen in plasma by scanning fluorometry after thin-layer chromatography

A rapid and sensitive method is described for determining 8-methoxypsoralen in plasma. Plasma samples are acidified with 6 mol/liter and heated in a boiling water bath to release the plasma-bound drug nondestructively. It then is extracted into a solvent mixture consisting of benzene/ethyl acetate (9/1 by vol). The solvent phase is separated, evaporated, and an aliquot of the dissolved residue is thin-layer chromatographed, with benzene/ethyl acetete (9/1) as developing solvent. The plate is dried and the spots, made visible under ultraviolet light (320-400 nm), are scanned. The smallest amount detectable is 20 ng; the overall analytical recovery from plasma is 84%. We used the method to determine the drug in the plasma of rabbits after intravenous and oral administration of 10 mg, and in one patient after an oral dose of 30 mg.