RESUMO
Saponins from Quillaja saponaria have been commonly used as immunomodulatory adjuvants in foot-and-mouth disease vaccines (FMDVs). However, due to the lack of consensus over the possible exacerbation of local inflammatory responses in cattle and its economic impacts, their use has been discouraged by Brazilian authorities. A qualitative method intended to determine the presence of saponins from Q. saponaria bark extracts in FMDVs was developed and validated. Instrumental analysis was performed using an liquid chromatography (LC) coupled to a quadrupole-time-of-flight-mass spectrometry (TOF-MS) system. The method was validated according to the International Conference on Harmonization Harmonized Tripartite Guideline Q2 (R1) and Brazilian Ministry of Agriculture, Livestock and Food Supply Analytical Quality Assurance Guidelines. Validation parameters were determined and considered suitable to the established criteria. The validated method has been applied in routine analysis in the National Agricultural Laboratory at Rio Grande do Sul (LANAGRO-RS). All results obtained were in agreement with the vaccine's composition described by the manufacturer. The method is easy and adequate for analysis in routine laboratories. To the best of the authors' knowledge, this is the first report of a method which intends to investigate the presence of saponins from Q. saponaria bark extracts in veterinary vaccines.
Assuntos
Adjuvantes Imunológicos/química , Cromatografia Líquida/métodos , Quillaja/química , Saponinas/análise , Vacinas Virais/química , Animais , Febre Aftosa/prevenção & controle , Espectrometria de Massas/métodos , Casca de Planta/química , Extratos Vegetais/química , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Background: Foot-and-mouth vaccines are an important tool in the control and eradication of the disease. In order to be commercialized, vaccines produced in Brazil undergo an evaluation process by health authorities, which includes sterility testing, residual active virus, potency, thermal stability, volume and non-structural protein activity. Sterility tests described in the Brazilian Pharmacopeia and by the World Organization for Animal Health (OIE) include direct inoculation and membrane filtration methods. The objective of the present study was to evaluate these two methods used to analyze sterility of vaccines against foot-and-mouth disease produced in Brazil. Materials, Methods & Results: Vaccines produced by the six main laboratories in Brazil were initially tested for filtration capacity. The sensitivity of the two techniques was determined artificially contaminating vaccines using known bacterial concentrations. Vaccines (9 bottles) from the same manufacturer were inoculated with 5 mL of steady-state growths of Pseudomonas aeruginosa, Candida albicans, and Clostridium sporogenes to final concentrations of 0.1, 1 and 10 CFU/ mL and a final volume of 55 mL. Bottles were manually shaken for 1 min to complete homogenization of contents. Then, 10 mL of each flask were used in assessment of the direct inoculation method, and 10 mL were used to evaluate the membrane [...](AU)
Assuntos
Febre Aftosa/imunologia , Febre Aftosa/prevenção & controle , Vacinas/análise , Controle de Qualidade , Técnicas Microbiológicas/veterinária , Filtração por MembranasRESUMO
Background: Foot-and-mouth vaccines are an important tool in the control and eradication of the disease. In order to be commercialized, vaccines produced in Brazil undergo an evaluation process by health authorities, which includes sterility testing, residual active virus, potency, thermal stability, volume and non-structural protein activity. Sterility tests described in the Brazilian Pharmacopeia and by the World Organization for Animal Health (OIE) include direct inoculation and membrane filtration methods. The objective of the present study was to evaluate these two methods used to analyze sterility of vaccines against foot-and-mouth disease produced in Brazil. Materials, Methods & Results: Vaccines produced by the six main laboratories in Brazil were initially tested for filtration capacity. The sensitivity of the two techniques was determined artificially contaminating vaccines using known bacterial concentrations. Vaccines (9 bottles) from the same manufacturer were inoculated with 5 mL of steady-state growths of Pseudomonas aeruginosa, Candida albicans, and Clostridium sporogenes to final concentrations of 0.1, 1 and 10 CFU/ mL and a final volume of 55 mL. Bottles were manually shaken for 1 min to complete homogenization of contents. Then, 10 mL of each flask were used in assessment of the direct inoculation method, and 10 mL were used to evaluate the membrane [...]