Methodology for isolating drug metabolites from biological fluids by HPLC

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Abstract
This article presents a detailed account of the isolation of drug metabolites from bile by reverse‐phase HPLC. The systematic approach is illustrated for the metabolism of butoprozine, a new antianginal drug, in the dog. Separation utilizes isocratic and gradient elution. Detection of the metabolites in. the presence of endogenous bile components is accomplished by very selective radioactivity measurements, by less selective UV (single wavelength) detection, and by much more universal detection modes such as on‐line UV spectroscopy and off‐line mass‐ and nuclear magnetic resonance spectroscopy. The importance of the latter three techniques in purity analysis of isolated metabolites is illustrated.

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