Hecker W, Witthauer D, Staerk A
Sandoz Pharma Limited, Basle, Switzerland.
PDA J Pharm Sci Technol. 1994 Jul-Aug;48(4):197-204.
Bacterial endotoxins (ETs) are lipopolysaccharides from the cell wall of Gram negative bacteria. ETs get into the environment as a result of autolytic desintegration of the bacterial cells. There exist a number of depyrogenation methods, either serving to remove or to inactivate ET. The most common means of ET inactivation is dry heat. Unfortunately no uniform regulation exists describing the conditions for sufficient ET inactivation. While the USP and FDA require an ET reduction of 3 log steps, no European regulation exists regarding depyrogenation of final containers for parenterals. However, the Ph. Eur. specifies the temperature and time conditions for depyrogenation of glassware in the pyrogen test monograph, allowing to choose between the two variants 250 degrees C/30 minutes or 200 degrees C/60 minutes which are not equivalent. In the present study those conditions for depyrogenation of glass containers in production of parenterals were investigated which, on the one hand, are technically feasible and, on the other hand, comply with the requirements of the main Pharmacopeias; furthermore, an ET preparation suitable for validation studies was selected. The preparation of ET indicators, the dry-heat inactivation and the recovery of ET are described in detail. Based on the results obtained, it is recommended to follow a defined treating temperature and period for safe depyrogenation of glass containers for parenterals, which results in a 4 log step reduction in ET without fillers. Thereby the USP/FDA requirement for a 3 log step reduction as well as the 200 degrees C/60 minutes requirement variant given in the Ph. Eur. can be fulfilled.
细菌内毒素（ETs）是革兰氏阴性菌细胞壁中的脂多糖。由于细菌细胞的自溶分解，内毒素进入环境。存在多种去热原方法，用于去除或使内毒素失活。最常见的内毒素失活方法是干热。不幸的是，不存在统一的法规描述充分使内毒素失活的条件。虽然美国药典（USP）和美国食品药品监督管理局（FDA）要求将内毒素降低3个对数级，但欧洲没有关于注射剂最终容器去热原的法规。然而，欧洲药典（Ph. Eur.）在热原检查专论中规定了玻璃器皿去热原的温度和时间条件，允许在250℃/30分钟或200℃/60分钟这两种不等效的变体之间选择。在本研究中，研究了注射剂生产中玻璃容器去热原的条件，这些条件一方面在技术上可行，另一方面符合主要药典的要求；此外，还选择了一种适用于验证研究的内毒素制剂。详细描述了内毒素指标的制备、干热失活和内毒素的回收率。根据获得的结果，建议遵循确定的处理温度和时间，以确保注射用玻璃容器安全去热原，这可使无填充物的玻璃容器内毒素降低4个对数级。从而可以满足USP/FDA要求的3个对数级降低以及欧洲药典中给出的200℃/60分钟要求变体。
