基于微量量热法的无菌制剂快速无菌检查方法研究

Rapid detection of microbial contamination of sterile preparation based on microcalorimetry

目的建立快速灵敏准确的无菌检查新方法,克服现有无菌检查方法检查周期长（14-21 d）、灵敏度低、自动化程度不高、容易引起误判等局限。方法采用微量量热法,以常规方法为参照,对方法灵敏度进行考察,建立基于微量量热法的无菌检查新方法;考察该方法对复方茵陈注射液正常样品、未灭菌样品、灭菌不彻底样品的检查能力。结果微量量热无菌检查法能够在10 h内检出各阳性菌（金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌、枯草芽孢杆菌、生孢梭菌、白色念珠菌、黑曲霉）,检出限度〈100 cfu,并能够快速检出模式药物未灭菌样品与灭菌不彻底样品,较常规检查方法更为灵敏快速。结论建立的微量量热无菌检查法灵敏、快速、准确,可实现自动在线监测,可以作为无菌检查的新方法。

Objective To establish a rapid and sensitive sterility test method,so as to overcome the limitations of long detection cycle（ generally 14- 21 d）,low sensitivity,low degree of automation,and prone misjudgement in conventional sterility test. Methods Microcalorimetry was adopted to establish a novel sterility test and compare with the conventional sterility test. Methodology investigation was carried out to ascertain the sensitivity of microcalorimetric sterility test. The discrimination capability of the microcalorimetric sterility test to the norm-sterilized samples（ Norm-SS）,non-sterilized samples（ Non-SS） and sub-sterilized samples（ Sub-SS） of Fufang Yinchen injections was further investigated. Results The results showed that microcalorimetric sterility test could detect out the positive indicator bacteria such as Staphylococcus aureus（ S. aureus）,Escherichia coli（ E. coli）,Pseudomonas aeruginosa（ P. aeruginosa）,Bacillus subtilis（ B. subtilis）,Clostridium sporogenes（ C. sporogenes）,Candida albicans（ C. albicans） and Aspergillus niger（ A. niger） within 10 h with the detection limitation of 〈100 cfu,and could distinguish the Non-SS and SubSS from Norm-SS accurately,rapidly and sensitively. Conclusion The microcalorimetric sterility test method is more sensitive,rapid and sensitive than the conventional culture medium turbidity observation method,and automatic online monitoring is possible.