Objective  To study the composition and distribution rules of microbial community in clean workshops of pharmaceutical enterprises after ozone disinfection, analyze the effect of ozone disinfection and sterilization, and provide data reference for the same space disinfection method of pharmaceutical enterprises.

Methods  Using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and VITEK^® 2 Compact identification methods, the collected planktonic and sedimentary bacteria were identified and analyzed.

Results  776 strains of bacteria were identified at the genus level, with 427 strains before disinfection and 339 strains after disinfection. The reduction rates of A, B and C enterprises were 22%, 19% and 23%, respectively. Before and after disinfection, the dominant bacterial genera were Staphylococcus, Bacillus, and Micrococcus. Micrococcus and Clostridium showed the most obvious disinfection effect, while Bacillus had the worst disinfection effect. The proportion of reduced bacterial contamination in the personnel changing area was the lowest, and company B could still collect contaminated bacteria in the A-grade filling area after disinfection, indicating that ozone disinfection didn't fully cover the potential risk of microbial contamination.

Conclusion  Ozone disinfection is a common method for disinfecting clean workshop spaces. After disinfection, the microbial population is regularly analyzed, and the disinfection level is further improved to ensure the effectiveness and safety of drug production.

1.刘亚茹, 余萌, 肖璜, 等. 五家制药企业洁净区微生物群落分析[J]. 中国药事, 2019, 33(7): 796-802. [Liu YR, Yu M, Xiao H, et al. Analysis of microfora in clean areas of five pharmaceutical enterprises[J]. Chinese Pharmaceutical Affairs, 2019, 33(7): 796-802.] DOI: 10.16153/j.1002-7777.2019.07.012.

2.U.S. Food and Drug Administration. Guidance for industry: sterile drug products produced by aseptic processing-current good manufacturing practice[EB/OL]. (2004-09-01) [2024-12-05]. http://www.fda.gov/media/71026/download．

3.European Commission. EU guideline to good manufacturing practice medicinal products for human and veterinary use. Annex 1: manufacture of sterile medicinal products (The second targeted  consultation) [EB/OL]. (2020-02-20) [2024-12-05]. https://ec.europa.eu/ health/sites/health/ files/files/ gmp/2020_annex1ps_sterile_medicinal_products_en.pdf．

4.Pharmaceuticals and Medical Devices Agency. Guidance on the manufacture of sterile pharmaceutical products by aseptic processing[EB/OL]. (2011-04-01) [2024-12-05]. http://www.pmda. go.jp/ files/000153543.pdf．

5.中国药典2020年版. 四部[S]. 2020: 503-506.

6.郭琦. 基于VITEK MS及VITEK2 Compact的更年宁微生物污染调查[J]. 中成药, 2024, 46(10): 3523-3528. [Guo Q. Investigation of microbial pollution in geng nian ning based on VITEK MS and VITEK2 compact[J]. Chinese Traditional Patent Medicine, 2024, 46(10): 3523-3528.] DOI: 10.3969/j.issn.1001-1528.2024.10.057.

7.宋明辉, 李琼琼, 秦峰, 等. 无菌药品生产企业过程污染微生物菌株库的建立及数据分析[J]. 药物分析杂志, 2019, 39(11): 1954-1960. [Song MH, Li QQ, Qin F, et al. Establishment and data analysis of a contaminated microorganisms library for sterile pharmaceutical production processes[J]. Chinese Journal of Pharmaceutical Analysis, 2019, 39(11): 1954-1960.] DOI: 10.16155/j.0254-1793.2019.11.05.

8.郑小玲, 王征南, 王知坚, 等. 药品微生物检测实验室环境菌库的建立[J]. 中国现代应用药学, 2015, 32(7): 847-850. [Zheng XL, Wang ZN, Wang ZJ, et al. Establishment of the environmentally microbial library in the drug sterility testing laboratory[J]. Chinese Journal of Modern Applied Pharmacy, 2015, 32(7): 847-850.] DOI: 10.13748/j.cnki.issn1007-7693.2015.07.019.

9.甘永琦, 阮斌, 零文超, 等. 药品企业生产车间洁净环境沉降菌的鉴定分析[J]. 药物分析杂志, 2020, 40(9): 1684-1691. [Gan YQ, Ruan B, Ling WC, et al. Identification and analysis of sedimentary bacteria of clean environment of production workshop in pharmaceutical manufacturers[J]. Chinese Journal of Pharmaceutical Analysis, 2020, 40(9): 1684-1691.] DOI: 10.16155/j.0254-1793. 2020. 09.21.

10.李南, 孙丽媛, 关奕泽, 等. 药物生产车间污染微生物的快速鉴定和污染微生物资源及信息库的建立[J]. 药物分析杂志, 2021, 41(3): 474-483. [Li N, Sun LY, Guan YZ, et al. Rapid identification of contaminated microorganisms in drug production work area and establishment of contaminated microorganism resource bank and database[J]. Chinese Journal of Pharmaceutical Analysis, 2021, 41(3): 474-483.] DOI: 10.16155/j.0254-1793.2021.03.13.

11.Sandle T. A review of clean room microflora: types, trends, and patterns[J]. PDA J Pharm Sci Technol, 2011, 65(4): 392-403. DOI: 10.5731/pdajpst 2011.00765.

12.张弛, 张建玲, 薛梅, 等. 制药企业CNC区域微生物数据库的建立及微生物种群分析[J]. 中国生物制品学杂志, 2023, 36(12): 1503-1514. [Zhang C, Zhang JL, Xue M, et al. Establishment of CNC regional microbial database and microbia population analysis for pharmaceutical enterprises[J]. Chinese Journal of Biologicals, 2023, 36(12): 1503-1514.] DOl: 10.13200/j.cnki.cjb.000082.

13.曹越, 帖金凤, 晁思琪, 等. 一种新型板式臭氧消毒机对空气及物体表面消毒效果研究[J]. 中国消毒学杂志, 2024, 41(3): 179-182. [Cao Y, Tie JF, Zhao SQ, et al. Study on disinfection effect of a novel plate ozone disinfector on air and object surface[J]. Chinese Journal of Disinfection, 2024, 41(3): 179-182.]DOI: 10.11726/j.issn.1001-7658.2024.03.006.

14.张彭义, 李惠南, 杨洁, 等. 臭氧用于空气和表面消毒的研究进展[J]. 安全与环境学报, 2021, 21(3): 1277-1291. [Zhang PY, Li HN, Yang J, et al. Research progress on ozone for air and surface disinfection[J]. Journal of Safety and Environment, 2021, 21(3): 1277-1291.] DOI: 10.13637/j.issn.1009-6094.2020.0848.

15.国家市场监督管理总局, 国家标准化管理委员会. 臭氧消毒器卫生要求. GB28232-2020[S]. 2020.