Reliable on-line sample preparation of basic compounds from plasma using a reversed phase restricted access media in column-switching LC.

We investigated on-line sample preparation of basic compounds from plasma using a methylcellulose-immobilized reversed-phase restricted-access media in column-switching liquid chromatography (LC). Dilution of the plasma sample with phosphate buffered saline prevented or delayed the formation of fibrin clots at 4 degrees C and resulted in reproducible on-line sample preparation over a 30-h period. The use of an ion-pair reagent in the extraction LC enhanced recoveries of hydrophilic basic compounds. The ability of the methods to quantify compounds in plasma were validated and the method was successfully applied to the pharmacokinetic study of a hydrophilic basic compound injected into the bloodstream of rats.