枳壳细菌污染状况及溯源分析  

Bacterial contamination status and traceability analysis of Aurantii Fructus

作　　者：霍晓菲 刘卫德 刘绪平 章红 HUO Xiao-fei;LIU Wei-de;LIU Xu-ping;ZHANG Hong(Jiangxi Institute for Drug Control,NMPA Key Laboratory of Quality Evaluation of Traditional Chinese Patent Medicine,Jiangxi Province Engineering Research Center of Drug and Medical Device Quality,Nanchang 330029,China;Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西省药品检验检测研究院、国家药品监督管理局中成药质量评价重点实验室江西省药品与医疗器械质量工程技术研究中心,南昌330029 [2]江西中医药大学,南昌330004

出　　处：《药物分析杂志》2023年第2期324-333,共10页Chinese Journal of Pharmaceutical Analysis

基　　金：江西省重点研发计划项目(20192BBGL70052);国家自然科学基金(81960715)。

摘　　要：目的:考察枳壳果实、药材、饮片的微生物污染水平,并对其污染细菌进行分离鉴定、溯源。方法:参考2020年版《中华人民共和国药典》通则1108《中药饮片微生物限度检查法》,对枳壳中的需氧菌总数、霉菌和酵母菌总数、耐热菌总数以及控制菌进行检查,采用PCR技术、RP技术对枳壳中检出的细菌进行菌株鉴定并通过BN软件溯源。结果:各阶段的微生物数量需氧菌总数及霉菌和酵母菌总数排序均为切片饮片、果实、麸炒饮片、药材。10批检出耐胆盐革兰氏阴性菌,但均未超过1×10^(4)cfu·g^(-1),未检出大肠埃希菌和沙门菌。共鉴定菌株50株,其中芽孢杆菌是该药材中的优势菌种,检出条件致病菌阪崎克罗诺肠杆菌、恶臭假单胞菌、肺炎克雷伯菌等。结论:为降低饮片中微生物负载,可尝试通过控制润药工艺参数如温度、水活度等方法防止微生物大量繁殖。果实到药材假单胞菌存在有同源性,而该类菌可能因为不耐热在饮片中并未分离到;阪崎克罗诺肠杆菌仅在饮片中存在,且占比高达50%,该菌可能是加工过程中由生产设备或环境或水体污染所致;另外,值得注意的是麸炒后的3批饮片中均存在条件致病菌肺炎克雷伯菌,应注意控制该类微生物污染,以保障枳壳质量安全。Objective:To investigate the level of microbial contamination in the fruits,herbs and tablets of Aurantii Fructus,and to isolate,identify and trace the source of contaminated bacteria.Methods:According to the microbial limit test method of the fourth part of Chinese Pharmacopoeia(2020 edition),the total number of aerobic bacteria,fungi and yeast,heat-resistant bacteria and control bacteria in Aurantii Fructus were examined.The bacterial strains detected in Aurantii Fructus were identified by PCR and RP technology and traced by BN software.Results:The total number of microorganisms,aerobic bacteria,fungi and yeasts in each stage were as follows:sliced slices,the fruit,bran fried slices,medicinal materials.Cholestate-resistant gram-negativebacteria were detected in 10 batches,but none of them exceeded 1×10^(4)cfu·g^(-1).Escherichia coli and salmonella were not detected in 10 batches.A total of 50 strains were identified,among which Bacillus is the dominant strain in the herb.The conditional pathogenic bacteria such as Cronobacter sakazakii,Pseudomonas aeruginosa and Klebsiella pneumoniae were also detected.Conclusion:To reduce the microbial load in the tablets,attempts can be made to prevent microbial blooms by controlling the moistening process parameters such as temperature and water activity.Pseudomonas were present in the fruit and the medicinal material,but not in the slices which can be due to heat intolerance from isolating.Cronobacter sakazakii was only present in the tablets and accounted for up to 50%of the total,which might be caused by contamination of production equipment or environment or water during processing;in addition,it is noteworthy that the conditional pathogenic bacteria Klebsiella pneumoniae were present in all three batches of tablets after bran-frying Klebsiella pneumoniae,and attention should be paid to control this type of microbial contamination to ensure the quality and safety of Aurantii Fructus.

关 键 词：枳壳 微生物限度检查 同源性分析 Ribo Printer微生物基因指纹系统 16S rDNA 

分 类 号：R917[医药卫生—药物分析学]