脉动真空湿热灭菌法去除内毒素的验证效果分析

Analysis in verification effect of removing endotoxin by plsating vacuum moist heat sterilization

目的检测在生物制药和医疗行业中用脉动真空湿热灭菌法去除内毒素验证效果的可靠性。方法首先应做脉动真空灭菌器的性能确认(PQ)其中包括:空载保压试验、狄克(b-d)试验、满载热穿透试验、微生物挑战试验,确认灭菌器性能合格后再将被灭菌物品和内毒素指示剂按着满载热穿透试验的装载方式共同放入灭菌器腔室内进行132℃、55min、脉动±65kPa灭菌程序和132℃、75min、脉动±65kPa灭菌程序的监测。结果用凝胶法限量检查:132℃、55min、±65kPa程序灭菌后的内毒素指示剂按内毒素检查法均出现凝集反应呈阳性,判定为不合格;检测132℃、75min、脉动±65kPa灭菌程序的内毒素指示剂按内毒素检查法无凝集反应呈阴性,做五遍均为阴性,判定为合格。结论用脉动真空高压蒸汽灭菌法进行132℃、75min、脉动±65kPa灭菌程序来去除内毒素,符合中国药典三部(2015版)关于内毒素的检测规定(不小于1000单位的细菌内毒素至少下降3个对数单位),可做进一步的探讨和验证。

Objective To test the reliability of verification effect of removing endotoxin by pulsating vacuum moist heat sterilization in the biopharmaceutical and medical industries.Methods First of all,the performance qualification(PQ)of the pulsating vacuum sterilizer should be carried out including no-load holding pressure test,Dick(b-d)test and full-load heat penetration test.After the sterilizer was confirmed to be qualified,the sterilized articles and the endotoxin indicator were placed together in the sterilizer chamber according to the loading mode of the full load heat penetration test for sterilization procedure of 132℃,55 min and pulsation±65kpa and sterilization procedure of 132C,75 min and pulsation±65kpa.Results Limited inspection by gel method:after sterilization at 132℃,55min and±65kpa,all the endotoxin indicators showed positive agglutination reaction according to the endotoxin test method,which was judged as unqualified.The endotoxin indicator tested at 132℃,75min and pulse±65kpa sterilization procedure was negative for no agglutination according to the endotoxin test and was negative for all the five tests,which was considered qualified.Conclusion Pulsating vacuum and high-pressure steam sterilization method is used to remove endotoxin by sterilization procedure of 132℃,75min and pulsating±65kpa,which is in line with the provisions of the three Chinese pharmacopoeia(2015 edition)on the detection of endotoxin(At least 3 logarithmic units of bacterial endotoxin are decreased for units not less than 1000)and can be further discussed and verified.