Determination of amphetamine, norephedrine, and their phenolic metabolites in rat brain by gas chromatography.

A specific analytical procedure for the quantitation of amphetamine (I), norephedrine (III), and their amphoteric metabolites, p-hydroxy-amphetamine (II) and p-hydroxynorephedrine (IV), in biological samples using electron-capture gas chromatography (GC-EC) is described. The procedure utilizes the ion-pairing reagent, bis(2-ethylhexyl)phosphoric acid, which frees the amines from most contaminants and permits the efficient extraction of the amphoteric compounds (as acetates) from the aqueous solution. Amines I and III and acetylated amines II and IV were perfluoroacetylated prior to GC-EC analysis. Metabolism of I, II, and III in the rat brain was studied. Results indicate that both in vivo and in vitro amines I and III are p-hydroxylated to II and IV, respectively, and II is beta-hydroxylated to give IV. Norephedrine (III) was not detected as a rat brain metabolite of amphetamine (I).