title: Evaluating measurement uncertainty in the microbiological assay of vancomycin from methodology validation data.
abstract: Methodology validation and measurement uncertainty estimation are fundamental to obtain reliable results. The microbiological methods are widely used to determine antibiotic assay, as they permit evaluation of the analyzed antibiotic activity. A microbiological assay of vancomycin was performed with adoption of experimental design 5 x 1 (interpolation in 5-point standard curve assay) with final concentrations from 6.4 to 15.6 microg/mL (standards) and 10.0 microg/mL (sample). Bacillus subtitlis (ATCC 6633) was the microorganism used, with antibiotic medium No. 8 as base layer and inoculated layer. The plates were incubated at 37 degrees C for 18 h. The method adopted for the microbiological assay of vancomycin through agar diffusion was validated according to statistic results demonstrated for suitability of the method concerning linearity, precision, and accuracy. The estimated relative expanded uncertainty (4.3%) was considered adequate for this method purpose.
authors.affiliation: University of São Paulo, Faculty of Pharmaceutical Science, Department of Pharmacy, São Paulo, Brazil. feliperl@usp.br
authors.forename: Felipe Rebello
authors.initials: FR
authors.lastname: Lourenço
authors.forename: Telma Mary
authors.initials: TM
authors.lastname: Kaneko
authors.forename: Terezinha de Jesus Andreoli
authors.initials: Tde J
authors.lastname: Pinto
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis; D000890:Anti-Infective Agents / Q000737:chemistry; D001412:Bacillus subtilis / Q000378:metabolism*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D019411:Clinical Laboratory Techniques; D004305:Dose-Response Relationship, Drug; D008826:Microbial Sensitivity Tests / Q000379:methods*; D015233:Models, Statistical; D012015:Reference Standards; D015203:Reproducibility of Results; D012107:Research Design; D013696:Temperature; D035501:Uncertainty; D014640:Vancomycin / Q000032:analysis*
chemical_list: D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D014640:Vancomycin