无菌检验

Sterility Testing Methods， Direct Inoculation – Considerations ，
无菌检验方法： 无菌检验方法：直接接种法
Advantages 优点 o Direct immersion of medical devices 医疗设备直接浸泡 o Non-filterable products may be tested 可以检验不可过滤产品 o Less manipulations than open funnel method 与开放过滤法相比操作简单 Limitations 局限性 o Antimicrobial product activity may inhibit growth 抗生素残留会抑制微生物 o Intrinsic product turbidity 产品本身可能会引起浑浊 o Aseptic technique training and validation required 需要无菌技术培训和验证 o No volumes larger than 100 ml 检验量小于100ml o High risk of false positives 假阳性的风险高
Follows requirements described in the Standard Pharmacopeias
按照药典标准规定的操作流程 1 – FILTRATION 过滤样品
SAMPLE
2 – RINSING 冲洗
RINSING FLUID
S
RR
3 – MEDIA ADDING 加培养基
其他一些国际组织的法规规定/指导 o TGA Australia o FDA: Food and Drug Administration o EMEA: European Agency for the Evaluation of Medicinal Products o PDA: Parenteral Drug Administration o AAMI: Association for the Advancement of Medical Instrumentation o ISO: International Organization for Standardization o WHO: World Health Organization
孔径不得大于0.45 µm
o Membrane diameter approximately: 50 mm
膜片直径约为50 mm
o Flow rate of water per minute / per cm², at a pressure of 13,5 PSI (70 cm de HG): 55 – 75 ml
Membrane Filtration:SteriSecure Closed concept system
薄膜过滤法： 薄膜过滤法：SteriSecure封闭式无菌检验系统 封闭式无菌检验系统 SteriSecure
Leader in this field (40 years of experience in sterility testing)
预灭菌的即用型产品
Allows to control the sample in the same way as at the time of his(her,its) use...
有很好的重现性和可重复性
How does SteriSecure work ? 如何操作SteriSecure 如何操作
该领域的领导者（40多年的无菌检验产品经验）
Method accepted from all Pharmacopoeias
所有药典都接受的方法
Fast filtration through pressure, not vacuum
通过加压而非抽真空进行快速过滤
Closed system
封闭系统
Pre-sterilised, ready-to-use units
从80年代早期开始：第一台隔离器（ La Calhène ）用于无菌检验
1983: specific membrane (Durapore, Millipore) for Antibiotics
1983：特殊材质的膜片(Durapore,Millipore)用于抗生素检测
1997: Incubation time for the test increased from 7 to 14 days
1. Hardware & Accessories 主机和配件
o o o Steritest Pumps Foot Pedal Timer (included in the Equinox)
2.
Sterility Testing Devices 无菌检验耗材
o Different Steritest Units with different adapters and membranes
Water 水系统
Environmental Monitoring 环境监控
Support Data
支持数据
Product Release 产品放行
Sterility Testing History 无菌检验发展历史
1957: membrane filtration (MF) for products with antimicrobial activity (FDA)
1977: Closed concept system Steritest (Millipore)
1977：发明了封闭式无菌检验体系Steritest（Millipore）
Since the early 80s: first isolators (La Calhène) for sterility testing
“对于每一批生成无菌和/或无热源的药品，都应有适宜的实验室检验确保其符合这些要求” 21CFR211.167 (a)
Example: EP
例如：欧洲药典
The test is applied to substances, preparations or articles which, according to the Pharmacopoeia, are required to be sterile (EP 5.3)
3.
Media & Rinse Fluids 培养基和冲洗液
o Recommended from regulatory agencies
Steritest Hardware 主机
Steritest Equinox for Laminar Flow Hood & Isolators 用于层流台和隔离器的Equinox 用于层流台和隔离器的
1997：检验的培养时间由7天增加到14天
Why to conduct a sterility test ? 为什么要进行无菌检验？ 为什么要进行无菌检验？
Regulations and global quality approach 法规和全球质量要求 cGMP requirements
cGMP的要求
根据药典规定，所有要求无菌的成分或制剂都必须进行检测（EP 5.3）
Why to conduct a sterility test ? 为什么要进行无菌检验？ 为什么要进行无菌检验？
European Pharmacopoeia (EP 5.3) 欧洲药典 US Pharmacopoeia (USP29-NF24) 美国药典 Japanese Pharmacopoeia (JP) 日本药典 ICH (International Committee of Harmonization) 国际协调会议 Regulatory Guidelines/Guidance from several organizations
4 – INCUBATION and Control 培 养和读数
M
2 MEDIA
MБайду номын сангаас
SteriSecure System Components for the test SteriSecure系统的组成 系统的组成
The Steritest System is a closed system to detect bugs in a sample by membrane filtration. Three Components:
“For each batch of drug product purporting to be sterile and/or pyrogen free, there shall be appropriate lab testing to determine conformance to such requirements” 21CFR211.167 (a)
Introduction to Sterility Testing
无菌检验
Yves Courties / May 2011
What is a sterility test ? 什么是无菌检验
Presence / Absence test 有/无检验 Reference test 法规要求的实验 Quality assurance requirement 质量保障的要求 Represents one set of data which contributes to the decision of whether or not the product lot meets the stated claims. 代表了一组数据，用来判别产品是否符合宣称要求 Not intended as a sole product release test 不是产品放行的唯一指标
Programs in Pharmaceutical Microbiology 制药行业微生物涉及的领域
Sterility Testing 无菌检验 Final Product 终产品
Microbiology Laboratory In-Process
制造过程
微生物实验室
Raw Materials 原材料
流速(在压力为13.5PSI时)：55-75ml/min/cm²
Sterility Testing Methods / Membrane filtration 无菌检验方法： 无菌检验方法：薄膜过滤法
(Open-) Funnel method 开放式方法
Closed concept system: Steritest 封闭式检测系统：Steritest
Sterility Testing Methods: Membrane Filtration 无菌检验方法： 无菌检验方法：薄膜过滤法
The US and European Parmacopoeias state: « Membrane filtration is the method of choice for sterility testing».
1957：膜过滤方法用于检测有抑菌性的产品（FDA）
1966: membrane filtration method improvement with filtration manifold (FDA). Usage of rinsing fluids
1966：膜过滤方法改进了过滤底座（FDA），并应用了冲洗液
美国和欧洲药典提到：薄膜过滤法是无菌检验可选的一种方法
Required conditions and specifications for a membrane filter destinated for sterility:
无菌检验对于过滤薄膜的要求：
o Pore size diameter (nominal) 0.45 µm maximum
Membrane Filtration (Open-) Funnel 开放式薄膜过滤法
Transfer, Filter and Rinse 转移，过滤，冲洗
Risk of False Positives !!!
Cut the filter 剪开膜片
假阳性的风险！ 假阳性的风险！
Transfer half of the filter in the media Incubate for 14 days 将膜片加入培养基培养 14天