Rapid and micro high-pressure liquid chromatographic determination of plasma phenytoin levels

A rapid and simple high-pressure liquid chromatographic microanalytical method was developed for the determination of clinically encountered plasma phenytoin levels. This method is accurate down to about 1 microgram of phenytoin/ml of plasma and requires as little as 10 microliter of sample. Total analysis time is about 10 min. The method involves deproteinizing with acetonitrile followed by monitoring the deproteinized sample at 254 nm. Phenytoin's primary metabolite in humans, 5-(p-hydroxyphenyl)-5-phenylhydantoin, also can be quantitated when present in moderately high clinically encountered concentrations. Plasma profiles of phenytoin and its metabolite were followed with time after an intravenous bolus injection to a rabbit.