Development and Validation of Europium-Sensitized Luminescence (ESL) Method for the Determination of Tetracycline Residu
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Development and Validation of Europium-Sensitized Luminescence (ESL) Method for the Determination of Tetracycline Residues in Milk Sara Morandi & Claudia Focardi & Mila Nocentini & Marta Puggelli & Gabriella Caminati
Received: 26 June 2008 / Accepted: 5 November 2008 / Published online: 26 November 2008 # Springer Science + Business Media, LLC 2008
Abstract The development of a fast and reliable screening method for the determination of tetracycline residues in milk is described and compared to high-performance liquid chromatography. The method is based on europium-sensitized luminescence: The emissive properties of the lanthanide cation are used as a fingerprint of europium (III) complexation with tetracycline. The optimization of the extraction and purification procedures provided a simple and convenient route to prepare stable tetracycline samples that can safely resist freezing cycles. The method is validated for the first time in milk matrix: The results show that accuracy and precision of the method meet complaints with the criteria of the Commission Decision 2002/657/EC. The proposed method has a false compliant rate (β error) lower than 5%. Keywords Tetracycline . Milk . Europium-Sensitized Luminescence . Screening Method . Validation
Introduction Members of the tetracycline antibiotics family, including tetracycline, oxytetracycline, chlortetracycline, and doxyS. Morandi : M. Puggelli (*) : G. Caminati (*) Dipartimento di Chimica, Universita’ di Firenze, Via della Lastruccia 3, 50019 Sesto Fiorentino, Italy e-mail: [email protected] e-mail: [email protected] C. Focardi : M. Nocentini Sezione di Firenze-Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana, Via di Castelpulci 41, 50010 San Martino alla Palma, Florence, Italy
cycline, are used worldwide as veterinary medicines and feed additives. The European Community is aware of the significant risk to consumer health due to the occurrence of veterinary drug residues in animal-derived food. The presence of antibiotic’s residues in food represents a risk for human health, and it should be correlated to the occurrence of strain resistance to antibiotics, allergic syndromes, and disorder of intestinal flora, with concomitant reduction of microbial production of vitamins. The Council Regulation (EEC) No. 2377/90 defines, for each molecule used in veterinary medicine, the maximum residue level (MRL) permitted in food for human consumption. In the specific case of tetracycline residues in milk matrix, a MRL of 100 μg/kg has been established. Tetracyclines have been detected in various biological matrices using high-performance liquid chromatography (HPLC) as confirmatory test, with different detection methods, i.e., spectrophotometry, electrochemistry, fluorescence, and mass spectroscopy (Loetanantawong et al. 2004; Pena et al. 2005). The specific aim of this work is to develop and validate a new screening method for the determination of tetracycline residues in milk matrix in order to provide an alternative to the ELISA method (Zhang et al.
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