Different agencies that supply validation guidelines worldwide establish almost the same parameters to be evaluated in the validation process of bioanalytical methods. However, they recommend different procedures, as well as establish different acceptance criteria. The present review delineates and discusses the stages involved in the validation procedures of bioanalytical methods designed for determining veterinary residues in food, explaining the main differences in the guidelines established for this purpose by the main regulatory agencies in the world.

This is an overview of LC-MS techniques applied for macrolide determination in food, including sample preparation and method validation, as well as the policies adopted by international agencies regarding their presence in food. Techniques for the analysis of macrolides in food normally include solid phase or liquid-liquid extraction followed by HPLC. UHPLC presents advantages in running time, detectability and solvent consumption. Triple-quadrupoles are the most common analyzers in instruments used for the determination of contaminants in food, but time-of-flight and ion-trap spectrometers have been successfully applied for analyses focusing on the investigation of structural formula or the presence of degradation products.

Brazil is one of the many countries that forbids the use of anabolic compounds, which generates difficulties on monitoring its use, once it has one of the biggest cattle herd. Therefore, several anabolic compounds are used, including trenbolone acetate. With the agreement of "Ministério da Agricultura, Pecuária e Abastecimento", an ELISA based test was done with the production of polyclonal antibodies in rabbits and testing in steers urine which received trenbolone acetate implants. The test showed to be cheap, easy and reliable to use in bovine urine to determine alfa- trenbolone, trenbolone acetate major metabolite, released in the animals urine until 60 days after implant was been injected. The results in comparison to the ones obtained by the commercial kit used by the Brazilian "Ministério da Agricultura, Pecuária e Abastecimento" were similar, with no significant differences.