Traditional methods for the determination of resorcyclic acid lactones, stilbenes and other compounds with hormonal activity in liver tissues have historically been time consuming and labour intensive. It is well known that veterinary drugs metabolise to form conjugates during their journey through a body but the parent compound needs to be released for analysis.  This has historically been achieved using an overnight enzymatic digest, and sample clean-up using liquid-liquid and solid phase extraction steps to give a sufficiently clean extract for analysis is performed after digest.<br/>A recently developed method using  an enhanced matrix  removal - Lipid extraction (EMR-Lipid) product has significantly decreased the time required for sample preparation. However, the requirement to determine the level of bound residue means that a time consuming digest step still needs to be included. By performing the enzymatic digest after the sample clean-up, the time required for digest has been significantly reduced, allowing for 24 samples to be extracted in a one-day method.<br/>The method was validated for the quantification of α- and β-trenbolone, zeranol, taleranol, zearalenone, zearalanone, α- and β-zearalenol, diethylstilbestrol, dienestrol and hexestrol in veal, bovine and equine liver.  Quantification limits are at or below 2.0 ng/g for all compounds in all species.<br/>A pilot study of 100 samples indicated that the method is suitable for routine use.<br/>

Development of a simple and rapid extraction method for the determination of resorcyclic acid lactones, stilbenes and trenbolone in liver tissues with enzymatic digestion

254th National Meeting (2017)

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