Desenvolvimento de métodos cromatográficos para o controle de qualidade e para a análise de resíduos em leite bovino de medicamentos veterinários
Abstract
This work reports the investigation of a series of bovine serum
albumin (BSA) restricted access media columns (RAM) for on-line milk samples deproteinization using water, phosphate and acetate buffer as mobile phase. The retention factor (k) of aminoglycosides antibiotics (streptomycin, neomycin and
gentamycin) on the RAM columns was evaluated using buffers and ion pair chromatography. The results showed a wide variation on aminoglicosides antibiotics retentions time; a RAM-ADS column was also investigated and the same results was observed. During the RAM columns studies, an investigation of two analytical columns, C18 and phenyl (Hyperil 15 x 0,46 cm i.d.), were performed using ion pair
chromatography. Perfluorinated acids (TFA, HFBA and PFPA) were evaluated as moblile phase and the detection was accomplished by an evaporative light scattering detector (ELSD). This study allowed the development of analytical
methods for quality control of gentamycin, neomycin and streptomycin in veterinary pharmaceutical formulations. The developed methods showed good selectivity, sensibility, accuracy and precision for all compounds allowing it to be
successfully used to commercials phormulation. This works also presents a study for on line determination of cephalosporinic antibiotics in bovine milk by multidimentional chromatography using RAM-BSA column for sample clean-up. The chromatographic conditions optimzed for these compounds were not adequate for the MS/MS quantification
due to suppression and/or enhancement of compound s ionization. Still a sensitive and enantioselective multidimensional high performance liquid chromatography (HPLC) method for determination of albendazole metabolites: albendazole sulphoxide, albendazole sulphone and albendazole 2-aminesulphone in bovine milk was development and validated at this work. The compounds, (±)-albendazole sulphoxide, albendazole sulphone and 2-aminesulphone were extracted from bovine milk using a octyl restricted access
media bovine serum albumin column (C8-RAM-BSA) (5.0 x 0.46 cm i.d.) and analyzed on a chiral column containing amylose tris(3,5-dimethylphenylcarbamate) coated onto APS-Nucleosil support, as stationary phase (15.0 x 0.46 cm i.d.). The
validated method showed linearity, precision, accuracy and adequate sensitivity, allowing it to be appropriate for pharmacokinetics studies and residual limits control. A matrix effect study was conducted to investigated its influence on the LC-MS method development.