Profiling of free and conjugated [3H]zeranol metabolites in pig plasma

Extraction and high-performance liquid chromatographic (HPLC) procedures are described that permit the complete analysis of free and conjugated zeranol metabolites in plasma from pigs implanted with [3H]zeranol. Free metabolites (9.0%) are extracted and then analysed by radio-HPLC on a reversed phase C18 column. They are distributed between three compounds that have been identified by gas chromatography-mass spectrometry as taleranol, zeranol and zeralanone. Direct radio-HPLC of the pre-extracted, deproteinated and Sep-Pak C18-purified plasma on a reversed-phase C18 column using tetrabutylammonium as an ion-pairing agent showed four main peaks: one corresponds to a weakly retained unidentified compound(s) (20%) and the other three were identified as the taleranol, zeranol and zeralanone glucuro conjugates. However, the total recovery is only about 25% owing to strong affinity of this polar material for the plasma proteins. Enzymatic deconjugation of the pre-extracted plasma followed by radio-HPLC analysis of the freed metabolites led to a good recovery of the radioactivity (81.8%) and allowed the quantitation of the different metabolites. These preliminary results indicate that zeranol is metabolized in the pig following pathways similar to those in other tested species.