关于蜡样芽孢杆菌片活菌数测定和杂菌检查方法的研究

Methods for Examining Viable Bacterial Count and Contaminating Bacteria of Bacillus cereus Tablets

目的对蜡样芽孢杆菌片现行标准中活菌数测定方法以及杂菌检查法的合理性进行探讨,并对这两法进行优化。方法对影响活菌数测定的因素,如培养基、稀释剂、稀释倍数、前处理方法,进行比较分析,选择较优的实验条件。使用微生态活菌制品总论下的杂菌检查法替代标准中的微生物限度检查法,并对其适用性进行研究。结果优化后的活菌数测定方法可以更加有效地复苏样品中的蜡样芽孢杆菌,结果明显高于现行标准方法。采用新的杂菌检查法能检出非致病性杂菌,发现样品可能存在风险,其适用性结果基本符合《中国药典》的要求。结论新方法适用于蜡样芽孢杆菌片活菌数测定和杂菌检查。现行标准中的活菌数测定法以及微生物限度检查不能很好地监测产品的质量风险,建议对方法进行修订,这样既可以更有效地监管产品,又可以更好地帮助生产企业提高产品质量。

OBJECTIVE To discuss the rationality of the current methods for examining viable bacterial count and contaminating bacteria in the standard of Bacillus cereus tablets,and optimize the two methods.METHODS The factors that affect the method for viable bacterial count were compared and analyzed,such as medium,diluent,dilution ratio,and pre-treatment,then the optimal experimental conditions were selected.The microbial limit test in the current standard was replaced with the contaminating bacteria test under the general chapter for probiotic in Chinese Pharmacopoeia part III,and its suitability was studied.RESULTS The optimized method for determination of viable bacterial count could more effectively recover Bacillus cereus in the sample,and the results were significantly higher than the current standard method.The use of a new method for detecting non-pathogenic bacteria can discover potential risks in the sample,and its suitability investigation results basically met the requirements of the Chinese Pharmacopoeia.CONCLUSION The new methods are suitable for the determination of viable bacterial count and detection of contaminating bacteria in Bacillus cereus tablets.The current standards for the determination of viable bacterial count and microbial limit testing cannot effectively monitor product quality risks.It is recommended to revise the methods.This can not only more effectively regulate products,but also better assist manufacturers in improving product quality.