A Multi-Dimensional Liquid Chromatography Method for Determination of Androgen Hormone Residues in Cattle Liver

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Abstract
A multidimensional HPLC method is described for the determination of trenbolone and 19-nortestosterone steroid hormone residues in cattle liver. Sample extracts are defatted by size exclusion chromatography. The evaporated residue is injected onto a high performance gel permeation column and the two fractions containing the steroids are diverted onto individual silica concentration columns, where the hormone residues are retained. The trapped hormones are then determined separately. Trenbolone held on one concentration column is eluted onto a cyano HPLC column and the steroid peak from this is transferred onto a final analytical silica HPLC column. After re-equilibration of the system the second concentration column is treated similarly for determination of 19-nortestosterone. Detection is by UV at 340 nm for trenbolone and 247 nm for 19-nortestosterone. Detection limits in cattle liver are 0.1 μg/kg for trenbolone and 0.3 μg/kg for 19-nortestosterone. For liver spiked at 2 μg/kg. recoveries of 59% and 39%, with CVs of 7% and 10% were obtained for trenbolone and 19-nortestosterone respectively. The low overall recovery for 19-nortestosterone was primarily attributable to losses during extraction.