ABSTRACT Veterinary drug residues in bulk tank milk are important to all sectors of the dairy chain because they are one of the major factors which determine the safety of the final product. This study attempted to identify milk quality parameters that are associated with the occurrence of veterinary drug residues using multivariate principal component analysis (PCA). A total of 132 raw milk samples were collected from 45 dairy farms in the state of Minas Gerais - Brazil and analyzed for 42 analytes, including pyrethroids, macrocyclic lactones and antibacterials, using liquid chromatography coupled with mass spectrometry in tandem mode and gas chromatography with electron capture detection. Out of the 132 milk samples, 40 samples tested positive for at least one analyte (above the detection limit). The milk parameters associated with the antimicrobial residues by confirmatory tests were lactose and nonfat concentrations, as revealed by PCA. This analysis showed that fat and total solid concentrations, as well as the somatic cell and total bacteria counts were associated with macrocyclic lactone residues in bulk tank milk. A PCA assessing pyrethroid residues in bulk tank milk revealed that the lactose and nonfat solid concentrations and titratable acidity were inversely associated with these residues. Thus, the data analysis indicated that the veterinary drug residues were associated with certain milk quality parameters that can be used to target farms at higher risk of veterinary drug residue contamination for testing programs in combination with incentives, education and training programs to improve mammary health, milk hygiene and safety.

RESUMO: Avaliou-se a presença de 42 analitos, incluindo piretróides, lactonas macrocíclicas e antimicrobianos em 132 amostras de leite de tanque proveniente de 45 propriedades leiteiras localizadas no Estado de Minas Gerais. Para tal, utilizou-se a cromatografia líquida acoplada a espectrofotometria de massas tandem e cromatografia gasosa com detector com captura de elétrons. Dentre todas as amostras de leite, 40 (30,30%) amostras de leite de tanque apresentaram a presença de pelo menos um analito, enquanto 16 amostras (12,12%) de leite demonstraram a presença de pelo menos dois analitos. Considerando os limites estabelecidos pela legislação brasileira, 11 amostras de leite (8,33%) seriam consideradas como não conforme. Ademais, os testes de triagem para detecção de antimicrobianos no leite não conseguiram identificar a maioria das amostras positivas nos testes confirmatórios, levando a grande discrepância entre estes testes. Desta forma, os resultados do presente estudo indicam que os períodos de descarte do leite, especialmente para piretróides, não foram plenamente respeitados por todos os produtores de leite. Além disto, uma discrepância entre os resultados dos testes confirmatórios e os testes de triagem foi observada.

ABSTRACT: A survey of veterinary drug residues in bulk milk tank from Minas Gerais State, Brazil, was carried out through a broad scope analysis. Here, 132 raw milk samples were collected at 45 dairy farms in Minas Gerais from August 2009 to February 2010, and analyzed for 42 analytes, comprising pyrethroids, macrocyclic lactones and antibacterials, using liquid chromatography coupled with mass spectrometry in tandem mode and gas chromatography with electron capture detection. Within all milk samples, at least one veterinary drug residue was identified in 40 milk samples (30.30%) by confirmatory tests, whereas 16 samples (12.12%) showed the presence of at least two residues. With regard to the Brazilian maximum residue levels, 11 milk samples (8.33%) were non-compliant according to Brazilian Legislation. The veterinary drugs detected in the non-compliant milk samples include penicillin V (one sample), abamectin (one sample) and cypermethrin (nine samples). Furthermore, the antibacterial screening methods failed to identify most of the positive samples that were detected by confirmatory tests, leading to a large discrepancy between the screening and confirmatory antimicrobial tests. Thus, the present study indicated that the veterinary drugs residues still represents a great concern for the milk production chain.

A procedure was developed for determination of 5 sedatives and 14 β-blockers in swine kidney and subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Three different procedures for extraction were tested, evaluated through recovery studies. The procedure using acetonitrile for extraction and cleanup with freezing at low temperature and dispersive solid phase extraction using 500 mg celite® 545 before the concentration step presented the better results. The dried samples were redissolved with methanol and analyzed using a LC-MS/MS system with electrospray ionization (ESI) operating in positive MRM mode. The recovery values for this procedure were in the 75-88% range. The robustness of the method was tested against small variations. The method was used to analyze carazolol, azaperone and azaperol in collaborative assay, obtaining results close to designed value.