Validating sterile product manufacturing processes requires the use of biological indicators to demonstrate the lethality of sterilization techniques. The indicator organisms are there merely to measure the effect of the sterilization process. Today, too much focus on lethality to biological mark-ers results in use of extreme sterilization conditions; insisting on excessive lethality requirements in sterile process validation threatens patient safety, the authors believe. Not only does the practice subject process equipment and raw materials to damaging conditions, it can lead drug manufacturers to opt for aseptic processing in cases where terminal sterilization would be the best and safest choice. Companies should be free to choose the best way to ensure a safe process, whether that is aseptic processing, terminal sterilization, or a combination of the two.
The control of microbial load on surfaces of research and animal labs as well as of hospital and pro-duction rooms is an everyday challenge. Surface sanitization or disinfection of production equip-ments and rooms by wiping with chemicals is a time-consuming procedure. Its validation is very intri-cate. Alternatively, fumigation allows overcoming many critical aspects of wiping in both procedure and validation. Moreover, the use of nebulized hydrogen peroxide (H(2)O(2)) of moderate concentra-tion resolves remaining reservations as to toxicity, corrosion, and persistence. This paper presents major criteria for the effective use of fumigation with emphasis on a new H(2)O(2) procedure. Focus is set to the proper and simplified validation of the process using standardized biological indicators with defined concentrations of the test organism Geobacillus stearothermophilus. Extensive test results from design of experiment studies support the knowledge of the sanitization process and allows for choosing critical process parameters within well known boundaries. Target values of total kill of 10(4) to 10(6) cfu per carrier are reproducibly reached. The new concept has then been applied to the decontamination of small rooms as well as to large multi-rooms up to 750 m(3). These case studies are described and illustrate the wide field of application of the technique proposed. The con-cept presented for validation of the sanitization procedure overcomes the problems associated with conventional surface disinfection validation. It allows for considerable more safety at greatly reduced cost and work.
The inactivation of endotoxin from six species of smooth gram-negative bacteria (S-form) by steam-heat treatment was investigated using the Limulus amebocyte lysate (LAL) assay. Biphasic decreases of endotoxins from four species of bacteria were observed upon steam-heat treatment of 1 mi-crogram/ml endotoxin solution at 121 degrees C in a steam sterilizer. A lag time, however, was ob-served in the inactivation profiles of V. cholerae and P. aeruginosa. Distinct differences in heat re-sistance were observed among the bacterial species. The decrease rate was found to be concentra-tion-dependent, and endotoxins at low concentrations (less than 10 ng/ml) were inactivated by the treatment to below the detection limit of the LAL assay. The time-course of the decrease of endotox-in from rough strains (R-form) resembled that of the respective S-form. The inactivation of R-form, especially Rc mutant, endotoxin was markedly affected by divalent cations such as Mg2+ and Ca2+, which appear to promote reaggregation of the endotoxin.