一例日沟维肠杆菌污染化妆品情况分析

doi:10.3969/j.issn.1001-1803.2023.04.006

摘要/Abstract

摘要：

按照《化妆品安全技术规范》（2015年版）对1批化妆品中菌落总数和6批化妆品中防腐剂进行检测；用VITEK MS微生物质谱检测系统和16s rRNA基因测序对化妆品中的污染菌进行鉴定；每月转接一次观察日沟维肠杆菌置常温条件下存活情况；用日沟维肠杆菌对5批次含羟苯甲酯的化妆品进行防腐剂挑战试验。结果显示，1批菌落总数超标的化妆品，其污染菌鉴定为日沟维肠杆菌，该批化妆品使用的防腐剂含量为0.15%羟苯甲酯。日沟维肠杆菌在常温营养条件较好的情况下可存活10个月以上。羟苯甲酯含量为0.2%的3批化妆品和羟苯甲酯含量为0.1%+双咪唑烷基脲1批化妆品通过了日沟维肠杆菌防腐剂挑战试验。羟苯甲酯含量为0.1%的化妆品未通过日沟维肠杆菌防腐剂挑战试验。日沟维肠杆菌对化妆品中常用防腐剂羟苯甲酯具有耐药性，选择该类防腐剂时推荐使用日沟维肠杆菌进行防腐剂挑战试验判定其适用性。

关键词: 日沟维肠杆菌, 防腐剂挑战试验, 羟苯甲酯, 16s rRNA基因测序

Abstract:

Enterobacter gergoviae is a conditional pathogen, which can cause urinary system, respiratory tract and blood infection in the immunocompromised population. It widely exists in water, soil and vegetables. Enterobacter gergoviae contamination is common in cosmetics, which is mainly related to its resistance to the most widely used cosmetic preservative methyl 4-hydroxybenzoate. In this paper, a batch of cosmetics contaminated by Enterobacter gergoviae were analyzed and studied in order to provide reference for the cosmetics enterprises to select appropriate preservatives. Aerobic bacterial was counted in 1 batch of cosmetics and the preservatives in 6 batches of cosmetics were tested according to the 2015 version of “Safety and Technical Standards for Cosmetics”. The contaminated bacteria in cosmetics were identified by VITEK MS and 16s rRNA gene sequencing, and transferred every month to observe the survival of Enterobacter gergoviae at room temperature. Enterobacter gergoviae was used to conduct preservative challenge test on 5 batches of cosmetics containing methylparaben. The results show that a batch of cosmetics with aerobic bacterial count exceeding the standard is identified as Enterobacter gergoviae, and the preservative methylparaben used in this batch of cosmetics is 0.15%. Enterobacter gergoviae can survive for more than 10 months under the condition of good nutrition at room temperature. Three batches of cosmetics with the methylparaben concentration of 0.2% and a batch of cosmetics with the methylparaben concentration of 0.1% + bisimidazolidinyl urea pass the preservative challenge test of Enterobacter gergoviae. Cosmetics with the methylparaben concentration of 0.1% fail the preservative challenge test of Enterobacter gergoviae. Enterobacter gergoviae is resistant to methylparaben, a commonly used preservative in cosmetics. When selecting this type of preservative, it is recommended to use Enterobacter gergoviae for preservative challenge test to determine its applicability.

Key words: Enterobacter gergoviae, preservative challenge test, methylparaben, 16s rRNA gene sequencing

中图分类号:

TQ658

黄依雯, 张珊珊, 王星月, 沈海英. 一例日沟维肠杆菌污染化妆品情况分析[J]. 日用化学工业（中英文）, 2023, 53(4): 408-413.

Huang Yiwen, Zhang Shanshan, Wang Xingyue, Shen Haiying. Analysis of a case of cosmetics contaminated by Enterobacter gergoviae[J]. China Surfactant Detergent & Cosmetics, 2023, 53(4): 408-413.

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参考文献 13

[1]	Nie Yanyan, Liu Donghong, Wu Huan, et al. Investigation on microbiological contamination of cosmetics available in guang dong market in 2015[J]. Flavour Fragrance Cosmetics, 2016 (6): 33-34.
[2]	Zhou Longxia. Investigation on microbial contamination of commercial cosmetics[J]. World Latest Medicine Information, 2016, 16(45) : 221.
[3]	Tang Huiqin, Wu Jianfei, Luo Jiao, et al. Biological characteristics of Pluralibactergergoviae[J]. Flavour Fragrance Cosmetics, 2021 (6): 32-35.
[4]	Davin-Regli P. Enterobacter gergoviae adaptation to preservatives commonly used in cosmetic industry[J]. International Journal of Cosmetic Science, 2014, 36(4): 386-395. doi: 10.1111/ics.2014.36.issue-4
[5]	Zhong Yu, Shi Di, Zhu Xiaona. Isolation and identification of a contaminating bacteria in eye cream[J]. Light Industry Science and Technology, 2018, 34(8): 126-127.
[6]	Wang Yongli. The microbial challenge test of several cosmetics[J]. Guangdong Chemical Industry, 2015, 42(3): 63-64.
[7]	Yang Juan, Liu Yonglong, Lin Rui, et al. Study on the inhibitory effect of several common preservatives on spoilage microorganisms in daily chemical products[J]. Industrial Microbiology, 2016, 46(4): 34-37.
[8]	Zhang Jing, Liu Xuping, Yi Luyao, et al. Analysis of microbial contamination and preservative use in cosmetics[J]. China Surfactant Detergent & Cosmetics, 2020, 50(10): 698-703, 710.
[9]	Zheng Ping, Ding Pei, Wu Liping, et al. Evaluation of p-hydroxybenzoate preservatives’s antibacterial effect[J]. Chinese Journal of Health Laboratory Technology, 2015, 25(10): 1540-1541, 1545.
[10]	Zhang Guolin, Su Yuanke, Shen Haiying, et al. Application of 16S rRNA/ITS sequencing and analysis in the identification, classification of microbial and drug quality control[J]. Chinese Journal of Modern Applied Pharmacy, 2017, 34(10): 1489-1495.
[11]	Sutton S, Jimenez L. A review of reported recalls involving microbiological control 2004—2011 with emphasis on FDA considerations of “objectionable organisms”[J]. Am. Pharm. Rev., 2012 (15): 42-57.
[12]	Luo Jun, Wang Xiaowei, Liu Yi, et al. Comparison and selection of effectiveness test methods of cosmetic antiseptic system[J]. China Surfactant Detergent & Cosmetics, 2021, 51(3): 243-249, 255.
[13]	Raymond C R, Paul J S, Paul J W. Handbook of pharmaceutical excipients[M]. Beijing: Chemical Industry Press, 2005.

检测时间	样品来源	菌落总数/（cfu/g）
2018年10月31日	1号和2号混合	1.9×10⁴
2018年11月05日	1号和2号混合	6.0×10⁵
2018年11月05日	3号和4号混合	1.0×10⁴
2018年12月29日	1号、2号、3号、4号	均为<10
2019年01月03日	1号、2号、3号、4号	均为<10

样品名称	与GenBank日沟维肠杆菌基因序列相似度/%
1号	99.45
2号	99.37
3号	99.62
4号	99.48
日沟维肠杆菌	99.72

样品序号	羟苯甲酯含量/%	菌落总数/（cfu/g）				结果
样品序号	羟苯甲酯含量/%	第0天	第7天	第14天	第28天	结果
A	0.1和双咪唑烷基脲	1.5×10⁶	0	0	0	通过
B	0.1	2.8×10⁶	4×10⁵	5.5×10⁵	1.6×10⁷	未通过
C	0.2	1.6×10⁶	0	0	0	通过
D	0.2	1.1×10⁶	0	0	0	通过
E	0.2	2.0×10⁶	0	0	0	通过

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