Resumo

Considering the setting of development of rapid microbiological methods, it is of fundamental importance to check whether the technique, among all advantages showed, have efficiency equal or superior to those already described in compendia, in order to obtain regulatory acceptability. Thus, this study is intended to assess the performance of a technology employing Adenylate Kinase-enhanced ATP bioluminescence in the sterility assessment of sterile pharmaceuticals, in comparison to the Sterility Test by Membrane Filtration, as defined in the official compendium. Tests using pilot batches of the following sterile products were performed: saline solution, polyelectrolyte concentrate for dialysis and metronidazole solution. Four microorganisms were evaluated: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis, and two concentration levels of microbial standards were used: N1 (20 CFU/filter membrane) and N2 (2 CFU/filter membrane). The procedure was performed in twenty-four samples for saline solution artificially contaminated with the suspension of each type of test microorganism in each one of the contamination levels, in twelve samples of polyelectrolyte concentrate for dialysis and other twelve samples of metronidazole solution. Two types of growth media and two incubation temperatures were utilized After 24, 48, 72, 96 and 120 hours and 7 and 14 days of incubation, aliquots of each culture medium were collected – from tests and controls – to be assessed by an alternative method (Celsis AKuScreen^TM protocol). And, after each aliquots collection, the culture media return to the incubation temperature conditions, being kept for 14 days until the final result for the sterility conventional test or until both methods returned a positive result. Results showed the Adenylate Kinase-enhanced ATP bioluminescence method can detect microbial growth within a period of 24 hours for 20 and 2 CFU/sample using the growth medium/temperature conditions evaluated. The data demonstrated applicability of the automated ATP bioluminescence system in detection of viable microorganisms in sterile products, compared with the Membrane Filtration method directed in USP XXXV.