组正交超饱和设计在发酵罐手工清洗方法开发中的应用

Application of group-orthogonal supersaturated designs in the development of the manual cleaning method of fermentation tanks

目的开发一种适用于发酵罐手工清洗方法。方法采用组正交超饱和设计方法(15个因素2个水平),试验因素包括9个实际影响发酵罐清洗因素和6个假因素;评价指标为发酵罐清洗后淋洗水电导率与总有机碳(total organic carbon,TOC)含量;通过标准最小二乘法拟合试验结果获得较优清洗条件,从而建立发酵罐手工清洗方法,进而通过试验验证发酵罐淋洗水电导率和TOC含量的实际值是否与预测值具有等效性。结果获得发酵罐手工清洗条件如下:预清洗次数1次,预清洗温度25℃;枸橼酸洗涤浓度5 g/L,枸橼酸洗涤次数1次,枸橼酸清洗温度25℃,酸洗涤剂去除清洗次数1次;氢氧化钠洗涤浓度5 g/L,氢氧化钠洗涤次数3次,碱洗涤剂去除清洗次数5次。验证试验淋洗水电导率和TOC含量检测结果均高于预测值,数据中位数均低于《中华人民共和国药典》2020版(四部)注射用水标准。结论此手工清洗方法对于发酵罐的清洗结果有效,且淋洗水检测结果理想。

Objective To develop a method for manual cleaning of fermentation tanks.Methods The group-orthogonal supersaturated design(GO-SSD)(15 factors and 2 levels)was adopted,and the experimental factors included 9 factors that actually affect the cleaning of the fermentation tank and 6 fake factors.The evaluation indexes were the conductivity and the amount of total organic carbon(TOC)in the rinse water from the fermentation tank cleaning.The optimal cleaning conditions were obtained to develop a manual cleaning method of fermentation tanks by the standard least squares method.Furthermore,it was verified by experiments whether the actual values of the conductivity and TOC content of the rinse water in fermentation tank were equivalent to the predicted values.Results The manual cleaning conditions of the fermentation tank were obtained as follows:the fermentation tank was pre-cleaned once at 25℃,and then washed once with 5 g/L citric acid at 25℃;the fermentation tank was washed once for removing acid detergent and then washed three times with 5 g/L sodium hydroxide solution;after that,the alkaline detergent was removed from the fermentation tank and this procedure was repeated five times.The verification test showed that the conductivity and TOC content of the rinse water were higher than the theoretical value,and the median of the test data was lower than the standards of water for injection in Pharmacopoeia of the People’s Republic of China 2020(volumeⅣ).Conclusion The manual cleaning method is effective for the fermentation tank cleaning and the test results of rinse water are ideal.