Beta-agonists in animal feed. II: Optimization of the extraction.

New projects of the European Commission, Measurement and Testing Programme (BCR) were set up in order to develop a modular sample preparation system for the determination of beta-agonists and animal feeds. Three phases are included: an extraction study, a clean-up study and finally a Second Intercomparison. This paper describes the extraction study in which four laboratories were involved. A total of 33 extraction conditions were tested regarding their yield on clenbuterol and salbutamol, their compatibility towards several clean-up and chromatographic end-methods and the influence of undesired coextractives. The conditions differed with respect to five factors: with or without organic solvent, temperature, pH, agitation and centrifugation. Their influence was examined via a ruggedness-test approach. A unique set-up allowed the combination of individual results in a complete factorial design. The addition of an organic solvent was found to be the most important factor. Interactions between factors were also studied. The best combinations of factors regarding the extraction are given. Finally limits for applicability and influence of organic solvents, pH and temperature were evaluated in a fifth laboratory towards enzyme immunoassay as detection method.