近日，PDA在PDA-letter上发表了关于胶塞斗的灭菌的文章——《对间接与产品接触的表面进行灭菌处理所面临的挑战》，该文章聚焦无菌生产中间接与产品接触的胶塞斗灭菌问题，对比可拆卸与不可拆卸胶塞斗的灭菌方式进行的风险分析并给出建议：

现状，33% 的企业胶塞斗不拆卸或过重，不满足欧盟 GMP 附录 1 要求，如使用替代程序需经质量风险管理验证。

符合欧盟药品生产质量管理规范（GMP）附件 1 的最简单方法是使用可拆卸的胶塞斗，并且新的灌装线应满足这一要求。或者，对不可拆卸部件的生产线进行重新设计也是一种选择，不过对于某些公司来说，这可能成本过高。每个制造商都应根据自身的操作程序和已实施的控制措施选择合适的程序。

 

原文及翻译如下

 

Challenges in Sterilizing Indirect Product-Contact Surfaces

 

对间接与产品接触的表面进行灭菌处理所面临的挑战

 

For aseptic processes, EudraLex GMP Annex 1: Manufacture of Sterile Medicinal Products, requires that direct and indirect contact parts are sterilized.

 

对于无菌生产工艺，欧盟药品管理法规《欧盟药品 GMP 附录 1：无菌药品的生产》要求对（与产品）直接和间接接触的部件进行灭菌处理。

 

Based on the current historical design of some filling lines, however, certain stopper bowls are either fixed, built-in or extremely heavy and not possible to remove for offline cleaning and sterilization. Facilitating the removal of these stopper bowls would require a significant redesign of existing lines. Therefore, a risk-based approach is needed to understand the risk of contamination during the preparation of the stopper bowl for aseptic production. Appropriate measures and controls should be developed and included in the company contamination control strategy document. This article proposes a risk assessment of two scenarios for the preparation of the stopper bowl—when it can be dismantled and when it cannot be dismantled. This article focuses on recommended procedures based on risk assessment and measures to mitigate the risk of contamination during aseptic production in both scenarios.

 

然而，基于目前一些灌装生产线的传统设计，某些胶塞斗要么是固定的、内置的，要么极其沉重，无法拆卸下来进行离线清洁和灭菌。要使这些胶塞便于拆卸，需要对现有的生产线进行重大重新设计。因此，需要采用基于风险的方法，来了解在为无菌生产准备胶塞的过程中存在的污染风险。应该制定适当的措施和控制方法，并将其纳入公司的污染控制策略文件中。本文针对胶塞的准备工作提出了两种情况的风险评估 —— 即胶塞能够拆卸和不能拆卸的情况。本文重点介绍了基于风险评估的推荐程序，以及在这两种情况下降低无菌生产过程中污染风险的措施。

 

Background

 

背景

 

According to a survey conducted in 2023 to more than 65 companies from around the world, 67% have a detachable stopper bowl, while the remaining 33% have a stopper bowl that is either not detachable or too heavy to be dismantled for cleaning and sterilization. Of these companies, 28% are located in the United States and Canada, 11% in Latin America (Brazil and Mexico), 8% in Asia (India, Korea, and Indonesia), 3% in South Africa and 36% in Europe (Belgium, Germany, Spain, Switzerland, Netherlands, Denmark, Ireland, Italy, the United Kingdom and France) (1).

 

根据 2023 年对来自全球 65 多家公司进行的一项调查，67% 的公司拥有可拆卸的胶塞（塞子），而其余 33% 的公司的胶塞要么不可拆卸，要么过于沉重，无法拆卸下来进行清洁和灭菌处理。在这些公司中，28% 位于美国和加拿大，11% 位于拉丁美洲（巴西和墨西哥），8% 位于亚洲（印度、韩国和印度尼西亚），3% 位于南非，36% 位于欧洲（比利时、德国、西班牙、瑞士、荷兰、丹麦、爱尔兰、意大利、英国和法国）（1）。

 

The survey shows that, as of 2023, there was a considerable percentage (33%) of manufacturers that do not comply with the requirements of the EU GMP Annex 1 2022 updates. For any newly installed filling lines, the expectation is clear that all direct and indirect product-contact parts should be able to be removed to be cleaned and sterilized offline. On the other hand, the implementation of alternative procedures differing from the recommendation in EU GMP Annex 1 will need to be justified with the tools of quality risk management (QRM) (2).

 

该调查显示，截至 2023 年，有相当比例（33%）的制造商不符合欧盟药品生产质量管理规范（GMP）附录 1 在 2022 年更新的要求。对于任何新安装的灌装生产线，明确的期望是所有与产品直接和间接接触的部件都应该能够拆卸下来，以便进行离线清洁和灭菌。另一方面，实施与欧盟 GMP 附录 1 中的建议不同的替代程序，需要使用质量风险管理（QRM）工具来证明其合理性（2）。

 

 

Figure 1 The survey indicates 67% of companies have a detachable stopper bowl and around 33% have stopper bowls which are either not detachable or too heavy to be washed out of place

 

图 1 该调查表明，67% 的公司拥有可拆卸的胶塞，大约 33% 的公司的胶塞要么不可拆卸，要么过于沉重，无法拆卸下来进行清洗。

 

The analysis below utilizes the Failure Mode and Effect Analysis risk-assessment tool to identify and assess the different risks associated with detachable and nondetachable stopper-bowl scenarios. The goal of this exercise will be to see what the major challenges in both are and what would be the best recommendations in each case. Many assumptions are made along the way to assess both situations, such as a certain facility design or a positive pressure isolator as barrier technology, and manufacturers might arrive at a different conclusion depending on the design of their facilities, barrier technologies, controls and procedures in place. Further, the analysis will be done for the stopper bowl as a challenging item due to its size, but it is applicable to any other indirect surface in the filling line.

 

下面的分析使用故障模式与影响分析风险评估工具，来识别和评估与可拆卸和不可拆卸胶塞情况相关的不同风险。这项分析工作的目标是了解这两种情况的主要挑战，以及在每种情况下的最佳建议。在评估这两种情况的过程中做了许多假设，例如特定的设施设计或作为隔离技术的正压隔离器，并且制造商可能会根据其设施设计、隔离技术、控制措施和现有程序得出不同的结论。此外，由于胶塞的尺寸原因，将其作为一个具有挑战性的项目进行分析，但该分析适用于灌装生产线中的任何其他间接表面。

 

Before starting with the assessment of the two scenarios, a brief introduction to risk management methodology and the different scales defined to assess the severity, probability and detectability will be presented.

 

在开始评估这两种情况之前，将简要介绍风险管理方法，以及为评估严重程度、发生概率和可检测性而定义的不同等级。

 

Risk Management Methodology

风险管理方法

 

Risk assessments consist of the (3):

 

风险评估包括以下内容（3）：

 

Identification of hazards

危害识别

 

Analysis

分析

 

Evaluation of risks associated with exposure to those hazards

对暴露于这些危害所相关的风险进行评估

 

To identify the risk(s) within the lifecycle of a stopper bowl, three questions are asked:

 

为了识别胶塞生命周期内的风险，需要提出三个问题：

 

What might go wrong?

可能会出现什么问题？

 

What is the likelihood (probability) it will go wrong?

出现问题的可能性（概率）有多大？

 

What are the consequences (severity)?

后果是什么（严重程度如何）？

 

The ability to detect the harm (detectability) was factored into the estimation of risk. The overall risk was quantitatively determined by multiplying the values of probability x severity x detectability for each identified risk. To quantify the overall risk, scales were defined to assess the severity, probability and detectability (3).

 

危害的可检测能力（可检测性）被纳入风险评估的考虑因素。通过将每个已识别风险的概率、严重程度和可检测性的值相乘，来定量确定总体风险。为了量化总体风险，定义了评估严重程度、概率和可检测性的等级（3）。

 

Severity

严重程度

 

The severity is defined as a measure of the possible consequences of a hazard (3). The scale proposed uses the following aspects to determine the level of severity:

 

严重程度被定义为对危害可能产生的后果的一种衡量标准（3）。所提出的等级标准利用以下几个方面来确定严重程度等级：

 

Probability of risk to product quality and to patient that could lead to partial or total batch rejection

对产品质量和患者存在的风险概率，这种风险可能导致部分或整批产品被拒收

 

Presence of further steps that can help reduce the associated risk

是否存在有助于降低相关风险的后续步骤

 

Proximity to the final filling of product

 

距离产品最终灌装环节的接近程度

 

Table 1 Severity scale proposed for the risk assessment

 

表 1 风险评估所提出的严重程度量表

 

 

Probability

可能性

 

The probability scale is defined in Table 2. The evaluation should be based on:

 

可能性等级在表 2 中进行了定义。评估应基于以下方面：

 

Data collected through previous experience in the facility

通过工厂先前经验收集的数据

 

Data coming from environmental monitoring (EM) and particle monitoring

来自环境监测（EM）和颗粒监测的数据

 

Data from previous deviations and investigations

来自先前偏差情况和调查的数据

 

Capability assessment if there is enough data

如果有足够数据的话，进行能力评估

 

Historical knowledge and published literature

历史知识和已发表的文献

 

Level of automation—In case of a manual step, how many manual steps are needed and what is the complexity of the manual steps to complete the task?

自动化水平 —— 如果是手动步骤，完成任务需要多少手动步骤，以及手动步骤的复杂程度如何？

 

As defined in Table 2, a rating of 1 means that, during the activity evaluated, it is not likely that microbial, particle or chemical contamination will occur. The type of contamination will depend on the step evaluated. In a cleaning step, for example, the risk is mainly of chemical contamination or cross-contamination. To assess the probability, if there is enough data for a capability analysis, this can be applied. If this is not the case, the scale in Table 2 suggests other options for the evaluation, such as by the level of automation, experience or published literature.

 

如表 2 中所定义的，评分为 1 意味着在评估的活动过程中，不太可能发生微生物、颗粒或化学污染。污染的类型将取决于所评估的步骤。例如，在清洁步骤中，风险主要是化学污染或交叉污染。为了评估可能性，如果有足够的数据进行能力分析，就可以采用这种方式。如果没有足够数据，表 2 中的等级给出了其他评估选项，比如依据自动化水平、经验或已发表的文献来评估。

 

For example, we can evaluate the cleaning step for the stopper bowl considering a common, yet challenging, residue like silicone oil.

 

例如，我们可以考虑像硅油这样常见但具有挑战性的残留物，来评估胶塞的清洁步骤。

 

If the stopper bowl is fixed, the cleaning step is done manually. According to Table 2, the probability can be assigned with a rating of 3 or even of 5 depending on how complex this activity is to be performed. This will depend on the size of the bowl and the design of the line.

 

如果胶塞是固定的，清洁步骤需手动完成。根据表 2，根据这项活动执行的复杂程度，可能性的评分可以是 3，甚至是 5。这将取决于胶塞的大小和生产线的设计。

 

If the stopper bowl is detachable, the probability of having issues with the cleaning is lower. The cleaning is usually done in a parts washer using a validated process. The use of laboratory coupon studies can help in designing a robust cleaning process, which will ensure the removal of silicone oil. In this case, the probability of having an unsuccessful cleaning is a rating of 1.

 

如果胶塞是可拆卸的，清洁出现问题的可能性较低。清洁通常在零件清洗机中使用经过验证的工艺来完成。利用实验室试片研究有助于设计一个可靠的清洁工艺，这将确保硅油被清除。在这种情况下，清洁不成功的可能性评分为 1。

 

Table 2 Probability scale proposed for the risk assessment

表 2 风险评估所提出的可能性等级

 

Detectability

可检测性

 

The detectability has to do with the controls in place to detect process failures. Controls that can be in place in a cleanroom are:

 

可检测性与用于检测工艺故障的现有控制措施有关。洁净室中可能采用的控制措施包括：

 

Particle monitoring

颗粒监测

Microbial monitoring or EM

微生物监测或环境监测（EM）

Microbial monitoring through contact plate or settle plate

通过接触碟或沉降碟进行微生物监测

Supervision of operators during procedures

在操作过程中对操作人员进行监督

Four-eyes principle

双人复核原则（四眼原则）

Visual inspection

目视检查

Continuous monitoring of critical variables

对关键变量进行持续监测

Table 3 shows the proposed scale for detectability. The evaluation should be done based on how good the controls in place are to detect the risk at the moment something is not performed as expected. For example, during manual material transfer, the detectability can be assigned a rating of 5 since it is very hard to detect a failure in the process leading to microbial contamination.

 

表 3 展示了所提出的可检测性等级。评估应基于现有控制措施在检测某事物未按预期执行时风险的能力有多强。例如，在手动物料转移过程中，可检测性可以评定为 5 分，因为很难检测到导致微生物污染的工艺故障。

 

Table 3 Detectability scale proposed for the risk assessment

表 3 风险评估所提出的可检测性等级

 

Risk Priority Number

风险优先数

 

The risk priority number (RPN) can be calculated as the multiplication of the probability, severity and detectability of each step. Table 4 shows a proposed scale for the overall risk classification according to the obtained RPN. Based on this scale, the risk can be determined as low, moderate, or high. It can be noted that, from a certain RPN, control measures are needed to reduce the risk. By doing this, only the detectability is going to be impacted. The severity and probability of the risk of the step should remain the same unless a substantial change is done to the process step.

风险优先数（RPN）可以通过将每个步骤的可能性、严重程度和可检测性相乘来计算。表 4 展示了根据所获得的风险优先数（RPN）提出的总体风险分类等级。基于这个等级，风险可以被确定为低、中或高。需要注意的是，从某个风险优先数（RPN）来看，需要采取控制措施来降低风险。通过这样做，只有可检测性会受到影响。除非对工艺步骤进行重大更改，否则该步骤风险的严重程度和可能性应保持不变。

 

Table 4 Overall risk classification according to the risk priority number

表 4 根据风险优先数划分的总体风险分类

 

Considerations – Facility Design and Barrier Technology

注意事项 - 设施设计和隔离技术

 

To assess the risks in each scenario, each step of the lifecycle of the stopper bowl will be analyzed.

 

为了评估每种情况下的风险，将对胶塞斗生命周期的每个步骤进行分析。

 

For the purpose of explaining the risks and control measures associated with the stopper bowl within its lifecycle, Figure 2 displays an example of the configuration of a facility. There are two different layouts—with (A) and without (B) a double-door autoclave—that will only impact the way in which the transfer process of the stopper bowl and other indirect surfaces is performed in Scenario 1, as will be explained. There are dedicated carts for Grade D and Grade C areas. An isolator with automated vaporized hydrogen peroxide (VHP) biodecontamination is considered the barrier technology available.

 

为了解释胶塞在其生命周期内的风险和控制措施，图 2 展示了一个设施配置的示例。存在两种不同的布局 —— 配备（A）和不配备（B）双门高压灭菌器 —— 正如将要解释的那样，这只会影响在场景 1 中胶塞和其他间接表面的转移过程的方式。有专门用于 D 级和 C 级区域的推车。带有自动汽化过氧化氢（VHP）生物净化功能的隔离器被视为可用的隔离技术。

 

(A) Double-door autoclave

（A）双门高压灭菌器

Figure 2 Double-door autoclave

图 2 双门高压灭菌器

 

(B) Without double-door autoclave

B）无双门高压灭菌器

 

Figure 3 Two different cleanroom layouts area—with and without double-door autoclave

图 3 两种不同的洁净室布局 —— 配备和不配备双门高压灭菌器

 

Scenario 1: Stopper Bowl is Detachable

场景 1：胶塞是可拆卸的

 

A typical lifecycle of the stopper bowl for this scenario can be seen in Figure 4.

在这种场景下，胶塞的典型生命周期如图 4 所示。

 

Figure 4 Lifecycle of a detachable stopper bowl

图 4 可拆卸胶塞的生命周期

 

Cleaning of the Stopper Bowl

胶塞的清洁

 

If the stopper bowl is detachable, it is usually cleaned with a parts washer. This eliminates the risk of poor reproducibility, which must be considered for scenario 2 with non-detachable stopper bowl.

 

如果胶塞是可拆卸的，通常会使用零件清洗机进行清洗。这就消除了重现性差的风险，而对于场景 2 中不可拆卸的胶塞，就必须考虑这种风险。

 

The risk within the cleaning process is associated with:

 

清洁过程中的风险与以下因素有关：

 

How hard is the residue to clean?

残留物的清洁难度如何？

How is the cleaning procedure designed?

清洁程序是如何设计的？

The harder it is to clean the residues, the higher the risk of the cleaning process failing. Residues of silicone oil coming from siliconized stoppers are quite common, as are hard-to-clean residues due to their poor solubility in water. The use of formulated chemistries that have different components in the formulation, such as surfactants, chelants and binders, will provide more effective cleaning for hard-to-clean soils (4). An appropriate cleaning procedure, its design based on laboratory coupon studies to support the selection of the right chemistry, concentration, temperature and time, will reduce the risks of failed procedures (4). Further, silicone residue is hard to detect via visual inspection and often not detected by operators. Easier-to-clean and easier-to-detect residues might reduce the risk within this step.

 

残留物越难清洁，清洁过程失败的风险就越高。来自硅化胶塞的硅油残留物很常见，由于其在水中的溶解性差，这些残留物很难清洁。使用配方中含有不同成分（如表面活性剂、螯合剂和粘合剂）的化学制剂，将对难以清洁的污渍提供更有效的清洁（参考文献 4）。基于实验室试片研究设计适当的清洁程序，以支持对正确的化学制剂、浓度、温度和时间的选择，这将降低清洁程序失败的风险（参考文献 4）。此外，硅酮残留物很难通过目视检查发现，而且操作人员往往无法检测到。更容易清洁和更容易检测到的残留物可能会降低这一步骤中的风险。

Figure 5 Risk evaluation of the cleaning step of a detachable stopper bowl

 

图 5 可拆卸胶塞清洁步骤的风险评估

 

Sterilization Wrapping – Preparation of the Stopper Bowl for Steam Sterilization

灭菌包装 —— 胶塞的蒸汽灭菌准备

 

The risk of this step is highly dependent on the wrapping material used. The manipulation from operators can also lead to particle and microbial contamination, though it is considered that the operators are well-trained in aseptic processing. Thus, this risk is lower in comparison to contamination coming from the wrapping material.

 

这一步骤的风险在很大程度上取决于所使用的包装材料。尽管认为操作人员在无菌操作方面训练有素，但操作人员的操作也可能导致颗粒和微生物污染。不过，与来自包装材料的污染相比，这种风险较低。

 

If Tyvek is used, the risk of particles is considered low, since peeling open a sealed Tyvek pouch generates less than one-tenth the number of particles (0.5 µ and 5 µ in size) compared to a cellulose pouch with a rating of 5. The probability of risk of particles would be a rating of 1. Severity is considered a rating of 3, as this can affect the product quality although not as severe as for a product rejection. Detectability is considered as a rating of 3 through particle monitoring, for example, giving an overall RPN=9, which falls in the green area.

如果使用特卫强（Tyvek）材料，颗粒污染的风险被认为较低，因为与评级为 5 的纤维素包装袋相比，打开密封的特卫强包装袋产生的颗粒（尺寸为 0.5 微米和 5 微米）数量不到其十分之一。颗粒污染风险的可能性评分为 1。严重程度评分为 3，因为这可能会影响产品质量，尽管不像导致产品被拒收那样严重。通过颗粒监测，可检测性评分为 3，因此风险优先数（RPN）为 9，处于绿色区域。

If cellulose material is used, the probability of particle contamination is considered as a rating of 3. This would lead to an RPN=27, which falls in the yellow area, corresponding to a moderate risk according to Table 4.

如果使用纤维素材料，颗粒污染的可能性评分为 3。这将导致风险优先数（RPN）为 27，处于黄色区域，根据表 4，对应中等风险。

Figure 6 Risk evaluation of the steps of preparation of the stopper bowl for sterilization and the sterilization process

图 6 胶塞灭菌准备步骤以及灭菌过程的风险评估

Sterilization of the Stopper Bowl in Autoclave

胶塞在高压灭菌器中的灭菌

 

A validated steam sterilization process with a well-maintained autoclave carries a low risk, as it is considered that a Bowie-Dick Test is used every day before using the autoclave. The risk in this step is also tightly associated with the wrapping material used. If the material considered is Tyvek, and there is data from the supplier on—

 

在维护良好的高压灭菌器中进行经过验证的蒸汽灭菌过程，其风险较低，因为通常认为在使用高压灭菌器之前每天都会进行鲍伊 - 狄克测试（Bowie-Dick Test）。这一步骤中的风险也与所使用的包装材料紧密相关。如果所考虑的材料是特卫强（Tyvek），并且供应商提供以下数据：

 

Good steam penetration

良好的蒸汽穿透性

Reproducibility during the wrapping process to ensure the same barrier to steam every time

包装过程中的可重复性，以确保每次对蒸汽具有相同的阻隔性

Good microbial barrier

良好的微生物阻隔性

Low particle generation

产生较少的颗粒

—then it is considered that the wrapping has a view window to allow the visual inspection of the parts for the rest of humidity after the cycle. The risk can be assessed as very low, giving an RPN=5.

 

那么可以认为这种包装有一个观察窗，以便在灭菌循环后目视检查部件是否残留水分。可认为该步骤风险非常低，风险优先数（RPN）为 5。

 

Transfer of the Sterilized Stopper Bowl from Grade D to Grade C

已灭菌胶塞从 D 级区域转移至 C 级区域

 

The transfer of materials into and out of the cleanrooms and critical zones is one of the greatest potential sources of contamination (2). The key to minimizing microbial and particulate contamination is to assess the possibilities available at the facility and look into the most practical and appropriate way to mitigate those risks. Different ways to perform material transfer include:

 

物料进出洁净室和关键区域的转移是最大的潜在污染源之一（参考文献 2）。将微生物和颗粒污染降至最低的关键在于评估工厂现有的可能性，并寻找最实际且合适的方法来降低这些风险。进行物料转移的不同方式包括：

 

Sterilization (double-door/ended) or depyrogenation tunnel

灭菌（双门）或除热原隧道

VHP or biodecontamination method

汽化过氧化氢（VHP）或生物净化方法

Removing outer layer of sterilized packaging

移除已灭菌包装的外层

Manual disinfection

手动消毒

The most practical way to minimize contamination risk is to use a double-door autoclave (2), but this is not always possible.

 

将污染风险降至最低的最实际方法是使用双门高压灭菌器（参考文献 2），但并非总是能够做到这一点。

 

The recommended sterilization wrapping for the stopper bowl when there is no double-door autoclave is a cover plus two sterilization-wrapping bags of flexible Tyvek. In the example in Figure 2 (B), the transfer of the stopper bowl will be done by removing the outer layer of sterilization wrapping. This method is preferred over manual disinfection due to the following:

 

在没有双门高压灭菌器的情况下，推荐用于胶塞的灭菌包装是一个外罩加上两个由柔性特卫强（Tyvek）制成的灭菌包装袋。在图 2（B）的示例中，胶塞的转移将通过移除灭菌包装的外层来完成。由于以下原因，这种方法优于手动消毒：

 

It eliminates a manual procedure with all its inherent challenges in reproducibility.

它消除了手动操作及其在可重复性方面的固有挑战。

The manual disinfection in this case is especially challenging since the outer sterilization wrapping layer is flexible and, therefore, difficult to cover with a wipe.

在这种情况下，手动消毒尤其具有挑战性，因为外层灭菌包装是柔性的，因此很难用擦拭布覆盖。

Removing an outer sterilization layer is faster than wiping with disinfectant, and it bypasses wet-contact times.

移除外层灭菌包装比用消毒剂擦拭更快，并且可以避免湿接触时间。

The recommended procedure for the transfer of the stopper bowl from Grade D to Grade C is:

 

将胶塞从 D 级区域转移至 C 级区域的推荐程序如下：

The cart is removed from the autoclave, and the parts are inspected for any condensation.

将推车从高压灭菌器中取出，并检查部件是否有任何冷凝现象。

The stopper bowl is placed on the cart side Grade D and driven into the material airlock.

将胶塞放置在 D 级区域的推车上，然后将推车驶入物料气闸。

With the help of another operator, and under continuous laminar airflow, the outer layer of wrapping is removed, and the stopper bowl is placed on the cart side Grade C.

在另一名操作人员的帮助下，在持续层流气流下，移除包装的外层，然后将胶塞放置在 C 级区域的推车上。

The stopper bowl remains with an inner layer of wrapping and can be stored or be directly installed in the filling line.

胶塞保留内层包装，可以储存或直接安装在灌装线上。

Without a continuous particle-monitoring in the material airlock, the step is ranked as moderate to high risk with an RPN=45. It is therefore recommended, if possible, to avoid having to do a manual material transfer and to automate the process with a decontamination chamber or a double-door autoclave.

 

由于在物料气闸中没有持续的颗粒监测，这一步骤被评为中高风险，风险优先数（RPN）为 45。因此，建议如果可能的话，避免进行手动物料转移，并通过净化室或双门高压灭菌器使该过程自动化。

 

Unwrapping and Installation of the Stopper Bowl in the Filling Line

胶塞在灌装线上的拆包和安装

 

This step poses a particular high risk when dealing with isolators, as the surrounding environment is a Grade C environment, for which the operators have a lower degree of gowning in comparison to a Restricted Access Barrier System (RABS), where the surrounding environment is Grade B. For this reason, to mitigate the risks during the unwrapping and installation of the stopper bowl, the operators should wear extra gowning such as face masks, goggles and Tyvek sleeves. On the other hand, isolators provide an “extra layer of protection” due to the biodecontamination with VHP, performed right before production starts. This helps mitigate any risk of contamination that could happen during the transfer of the stopper bowl and the unwrapping and installation process. In recent years, RABS has also employed automated VHP biodecontamination before production starts by applying VHP in the Grade B rooms and Grade A RABS at the same time. The recommended procedure would be:

 

在处理隔离器时，这一步骤具有特别高的风险，因为周围环境是 C 级环境，与限制进入屏障系统（RABS）相比，操作人员在 C 级环境中的着装要求较低，而在 RABS 中，周围环境是 B 级。因此，为了降低胶塞拆包和安装过程中的风险，操作人员应穿戴额外的防护装备，如口罩、护目镜和特卫强（Tyvek）袖套。另一方面，由于在生产开始前立即进行汽化过氧化氢（VHP）生物净化，隔离器提供了 “额外的防护层”。这有助于降低在胶塞转移以及拆包和安装过程中可能发生的任何污染风险。近年来，限制进入屏障系统（RABS）也在生产开始前通过同时在 B 级房间和 A 级 RABS 中应用汽化过氧化氢（VHP）来进行自动化的生物净化。推荐程序如下：

 

The wrapped stopper bowl should be transported with the cart until right before the limit between Grade C and Grade A areas (punctuation line on Figure 2). The unwrapping should happen inside the area between the punctuation line and the isolator doors. In this area, continuous laminar flow should be coming from the isolator Grade A area.

已包装的胶塞应随推车运输，直到 C 级和 A 级区域之间的界限处（图 2 中的标点线）。拆包应在标点线和隔离器门之间的区域内进行。在该区域，应从 A 级隔离器区域有持续的层流气流。

Continuously under the laminar flow, with the help of an operator, the outer cover of the stopper bowl should be unwrapped.

在持续的层流气流下，在一名操作人员的帮助下，应拆除胶塞的外罩。

Both operators should place the stopper bowl on the line for installation.

两名操作人员应将胶塞放置在生产线上进行安装。

The last cover can be left on the stopper bowl until after the VHP cycle and right before aseptic production starts.

胶塞的最后一层包装可以保留，直到汽化过氧化氢（VHP）循环结束且无菌生产开始前。

This transfer step is the most critical in this scenario with an RPN=30. The high number is due to a lack of continuous real-time EM that could detect any issues during the procedure. This requires relying on particle monitoring and aseptic process simulation to ensure the procedure is functioning as expected.

 

在这种情况下，这一转移步骤是最关键的，风险优先数（RPN）为 30。数值较高是因为缺乏能够在操作过程中检测到任何问题的持续实时环境监测（EM）。这需要依靠颗粒监测和无菌工艺模拟，以确保该操作按预期进行。

 

As defined in this example, an RPN=30 demands measures to be in place to control the risk such as:

 

正如本示例中所定义的，风险优先数（RPN）为 30 需要采取适当的措施来控制风险，例如：

 

Supervision by other operators

由其他操作人员进行监督

Good aseptic technique

良好的无菌技术

Well-written procedures

编写完善的操作程序

This should help in mitigating the risk in the transfer and installation step.

 

这将有助于降低转移和安装步骤中的风险。

Figure 7 Risk evaluation of the unwrapping and installation of the stopper bowl in the filling line图 7 胶塞斗在灌装线上拆包和安装的风险评估

Scenario 2: Stopper Bowl is Nondetachable

场景 2：胶塞不可拆卸

 

In this case, a manual cleaning and disinfection of the stopper bowl can be used as an alternative procedure. A typical lifecycle of the stopper bowl for this scenario can be seen in Figure 8.

在这种情况下，可采用对胶塞进行手动清洁和消毒的替代程序。这种场景下胶塞的典型生命周期如图 8 所示。

Figure 8 Lifecycle of a Nondetachable Stopper Bowl

图 8 不可拆卸胶塞斗的生命周期

Transfer and Preparation of Cleaning, Disinfection and Rinsing Agents

清洁、消毒剂的转移及准备

 

The cleaning and disinfecting agents used inside Grade A and Grade B areas should be sterile prior to use (2).

 

在 A 级和 B 级区域内使用的清洁和消毒剂在使用前应是无菌的（参考文献 2）。

 

The transfer of these materials will depend on the format in which they are packed. If the products used are ready-to-use (RTU) wipes, they come sterile and double-bagged. These can be transferred using the material airlock shown in Figure 2 by removing the first packaging layer inside the Grade D area and placing the wipes on the cart side of the Grade C area. The last packaging layer will be removed inside the isolator or in the material airlock of the isolator, prior to use of the wipes.

 

这些物料的转移取决于它们的包装形式。如果使用的产品是即用型（RTU）擦拭布，它们是无菌的且采用双层包装。可以使用图 2 所示的物料气闸进行转移，在 D 级区域内移除第一层包装，然后将擦拭布放置在 C 级区域的推车上。在使用擦拭布之前，应在隔离器内或隔离器的物料气闸中移除最后一层包装。

 

In case the product comes in spray form and is double-packed, the same procedure for material transfer can be performed. The spray and trigger may come separately; both are to be transferred in the same way. Right before starting the procedure, the trigger can be pushed into the spray bottle inside the material airlock of the isolator and then transferred inside the isolator.

 

如果产品是以喷雾形式且采用双层包装，则可以执行相同的物料转移程序。喷雾瓶和喷头可能是分开的，两者都应以相同的方式进行转移。在开始操作之前，应在隔离器的物料气闸内将喷头安装到喷雾瓶上，然后转移到隔离器内。

 

Sterile polyester wipes can be transferred either by removing the outer packaging layer or by disinfecting the packaging every time it is transferred to a higher classification. If the disinfection method is used, it should be performed in the lower classification area, while the wet-contact times should be observed in the higher classification area of the airlocks.

 

无菌聚酯擦拭布可以通过移除外包装层或在每次转移到更高等级区域时对包装进行消毒的方式进行转移。如果采用消毒方法，则应在较低等级区域内进行，同时应在气闸的较高等级区域内遵守湿接触时间的要求。

 

The risks associated with the preparation of the cleaning and disinfection (C&D) agents will increase if the use-dilution of detergent/disinfectant needs to be prepared and filtered under a laminar flow inside the Grade A area. The more manipulation needed for the preparation, the higher the risks for contamination and compromise of the sterile state of the agent used.

 

如果需要在 A 级区域内的层流条件下对洗涤剂 / 消毒剂进行使用稀释和过滤，那么与清洁和消毒剂（C&D）准备相关的风险将会增加。准备过程中需要的操作越多，所使用的试剂被污染以及破坏其无菌状态的风险就越高。

 

To evaluate the risks of this step, the following need to be considered:

 

为了评估这一步骤的风险，需要考虑以下因素：

 

Use of RTU wipes – low risk on preparation; otherwise, high risk

使用即用型（RTU）擦拭布 —— 准备过程中风险较低；否则，风险较高

Manual or automated material transfer (e.g., with a decontamination chamber)

手动物料转移或自动化物料转移（例如，使用净化室）

Controls in place during the material transfer, such as continuous particle-monitoring in the material airlock

物料转移过程中的控制措施，例如在物料气闸中进行持续的颗粒监测

For this example, the material transfer will be considered to be manual and RTU wipes are used. The probability of a risk happening during a manual material transfer is considered as having a rating of 3. Since there is usually no continuous particle monitoring or EM in place in a material airlock, it is difficult to detect if something goes wrong during the process. The only control in place is visual inspection. Detectability is assigned a rating of 5. The severity is classified as 2 because there is a VHP step prior to production to eliminate potentially introduced contamination. This step is identified as middle-to-high risk (RPN=30). If a decontamination chamber is available, the probability would decrease to 1, and the step would have an RPN of 10.

 

在本示例中，物料转移被认为是手动的且使用即用型（RTU）擦拭布。手动物料转移过程中发生风险的可能性评分为 3。由于在物料气闸中通常没有持续的颗粒监测或环境监测（EM），因此很难检测到过程中是否出现问题。现有的唯一控制措施是目视检查。可检测性评分为 5。严重程度分类为 2，因为在生产前有汽化过氧化氢（VHP）步骤来消除潜在引入的污染。这一步骤被确定为中高风险（风险优先数（RPN）=30）。如果有净化室，可能性将降至 1，该步骤的风险优先数（RPN）为 10。

 

Manual Cleaning of the Stopper Bowl

胶塞的手动清洁

 

It is important to understand the possible residues that need to be cleaned to select the right cleaning agent. Hard-to-clean residues such as silicone oil usually need an alkaline-formulated detergent. Other easier-to-clean residues might be cleaned using a neutral-formulated detergent, H2O2, 70% Isopropyl alcohol (IPA) or Water for injection (WFI). When cleaning with a formulated chemistry, WFI or 70% IPA applied in wipes can be used to remove surfactant residue.

 

了解需要清洁的可能残留物以选择合适的清洁剂是很重要的。像硅油这样难以清洁的残留物通常需要碱性配方的洗涤剂。其他较易清洁的残留物可以使用中性配方的洗涤剂、过氧化氢、70% 异丙醇（IPA）或注射用水（WFI）进行清洁。当使用配方化学制剂进行清洁时，可以使用沾有注射用水或 70% 异丙醇的擦拭布来去除表面活性剂残留物。

 

The manual cleaning of the stopper bowl will be done with the doors open, unless it is possible using another method. Besides the inherent risks of the manual cleaning itself, other measures are necessary to minimize the risk of particle and microbial contamination during the procedure, such as:

 

胶塞的手动清洁通常需要在门打开的情况下进行，除非可以使用其他方法。除了手动清洁本身固有的风险之外，还需要采取其他措施来将操作过程中颗粒和微生物污染的风险降至最低，例如：

 

Laminar flow working throughout the process

在整个过程中保持层流

Extra gowning, for example, face masks, goggles, and Tyvek sleeves

穿戴额外的防护装备，例如口罩、护目镜和特卫强（Tyvek）袖套

Practice of first cleaning nonproduct-contact areas or low-risk areas, followed by high-risk areas like the stopper bowl in the cleaning procedure.在清洁程序中，先清洁非产品接触区域或低风险区域，然后清洁像胶塞斗这样的高风险区域

As a critical indirect-surface area, the cleaning of the stopper bowl needs to be validated using visual inspection and both analytical and sampling methods. Analytical methods will depend on the residue to be traced. Total organic carbon (TOC) analysis can be challenging due to interference coming from IPAs commonly used in these areas. Therefore, HPLC and conductivity methods are preferred. Another challenge is the calculation of a cleaning limit for a stopper bowl where there is no direct contact with the product, but contact with a primary-packaging material.

 

作为关键的间接表面区域，胶塞的清洁需要通过目视检查以及分析和取样方法来进行验证。分析方法将取决于要追踪的残留物。由于这些区域通常使用异丙醇会产生干扰，因此总有机碳（TOC）分析可能具有挑战性。因此，更倾向于使用高效液相色谱（HPLC）和电导率方法。另一个挑战是计算与产品无直接接触但与初级包装材料接触的胶塞斗的清洁限度。

 

Manual cleaning introduces the usual risks associated with reproducibility and is more carefully reviewed by authorities. Measures that can be taken to minimize these risks are:

 

手动清洁会带来与可重复性相关的常见风险，并且会受到监管部门更严格的审查。可以采取以下措施来将这些风险降至最低：

 

Procedures should be easy to follow

操作程序应易于遵循

Standard operating procedures should be detailed, and operators should be trained frequently

标准操作程序应详细，并且操作人员应经常接受培训

Procedures should be supervised at a certain frequency or every time (four-eyes principle)

应按一定频率或每次都对操作程序进行监督（双人复核原则）

Monitoring procedures, like swabbing, to confirm the cleaning process was effective should be done after each cleaning or at least with a higher frequency as done in automated cleaning processes (6).

应进行监测程序，例如擦拭取样，以确认清洁过程是有效的，每次清洁后都应进行，或者至少像在自动化清洁过程中那样以更高的频率进行（参考文献 6）。

Some of these measures might be challenging to execute, depending on the production schedule, such as frequent training, observations of the process and periodic swabbing of the surface. Nonetheless, a compromise should be found in which the risks can be minimized, and resources are not exceeded.

 

根据生产计划，其中一些措施可能执行起来颇具挑战性，比如频繁的培训、对生产过程的观察以及定期的表面擦拭取样。尽管如此，还是应该找到一种折中的办法，既能将风险降至最低，又不会过度消耗资源。

 

As mentioned, RTU wipes not only simplify the material transfer but also eliminate the need to prepare the use-dilution inside the Grade A area, thereby minimizing risks and the number of manual activities overall. The use of laboratory coupon studies can help explore and define the right procedure such as how many wipes to use, how many strokes per wipe are needed and which kind of wiping pattern is optimal (e.g., unidirectional overlapping strokes, back and forth). Rinsing with 70% IPA to remove the excess of surfactants can also be simulated in the laboratory.

 

如前所述，即用型（RTU）擦拭布不仅简化了物料转移，还消除了在 A 级区域内准备使用稀释液的需求，从而总体上降低了风险并减少了手动操作的数量。使用实验室试片研究可以帮助探索和确定正确的操作程序，例如使用多少擦拭布、每次擦拭需要多少次擦拭以及哪种擦拭模式是最佳的（例如，单向重叠擦拭、来回擦拭）。还可以在实验室中模拟使用 70% 异丙醇进行冲洗以去除多余的表面活性剂。

 

The main risk during this step is not having a successful cleaning. There is a risk of introducing contamination during this intervention; however, manufacturers with experience in this type of cleaning report that the risk is very low. The probability of this risk should be revised and assessed based on historical data and the experience of each manufacturer as this will depend on the residue, the cleaning agents used, the procedure and the design of the filling line. For this example, the residue is considered to be silicone oil. The probability of an unsuccessful cleaning can be considered a rating of 3or even a5because this is a manual process, and silicone oil is a hard-to-clean residue. Due to the difficulties of assessing silicone residue via visual inspection, the detectability is assigned as a rating of5. Severity is assigned a2rating as this residue would not impact product quality. This gives an RPN between30and50, middle-to-high risk. The probability of this risk can be reduced to rating of3if a laboratory model is used to design the cleaning procedure. This would result in an RPN between10and30. If the residue is easy to clean, the probability of having an unsuccessful cleaning can be reduced to a 1, resulting in an RPN of10.

 

这一步骤的主要风险是清洁不成功。在这一干预过程中存在引入污染的风险；然而，在这种类型的清洁方面有经验的制造商报告说，这种风险非常低。应根据历史数据和每个制造商的经验来修订和评估这种风险的可能性，因为这将取决于残留物、所使用的清洁剂、操作程序和灌装线的设计。在本示例中，残留物被认为是硅油。清洁不成功的可能性可被认为评分为 3 甚至 5，因为这是一个手动过程，并且硅油是一种难以清洁的残留物。由于难以通过目视检查评估硅酮残留物，可检测性评分为 5。严重程度评分为 2，因为这种残留物不会影响产品质量。这使得风险优先数（RPN）在 30 到 50 之间，属于中高风险。如果使用实验室模型来设计清洁程序，这种风险的可能性可以降低到评分为 3。这将导致风险优先数（RPN）在 10 到 30 之间。如果残留物易于清洁，清洁不成功的可能性可以降低到评分为 1，风险优先数（RPN）为 10。

Figure 9 Risk evaluation of the transfer of cleaning and rinsing agents and the manual cleaning step of a nondetachable stopper bowl

图 9 不可拆卸胶塞的清洁和冲洗剂转移以及手动清洁步骤的风险评估

Manual Disinfection of Stopper Bowl

胶塞的手动消毒

 

A sporicidal agent is recommended to be used as disinfectant as this is an indirect surface near the final filling of the product. A sporicidal agent, such as a peracetic acid/H2O2 blend, will ensure full microbial inactivation. IPA 70% applied in wipes is recommended as the rinsing method to allow for evaporation after the rinse step is completed.

 

建议使用杀孢子剂作为消毒剂，因为胶塞是靠近产品最终灌装处的间接表面。像过氧乙酸 / 过氧化氢混合物这样的杀孢子剂能确保完全灭活微生物。建议使用沾有 70% 异丙醇的擦拭布作为冲洗方法，以便在冲洗步骤完成后使其蒸发。

 

As the surface of the stopper bowl is in direct contact with stoppers (primary product-contact surfaces), the stopper bowl requires cleaning validation. This means that that the rinsing step should be demonstrated as able to remove the sporicidal agent below a calculated cleaning limit. The sporicidal agent should have a Permitted Daily Exposure (PDE) or Acceptable Daily Exposure (ADE) value available, and the shared surface area should be considered for the calculation of the cleaning limit.

 

由于胶塞的表面与胶塞（主要的产品接触表面）直接接触，胶塞需要进行清洁验证。这意味着冲洗步骤应证明能够将杀孢子剂去除到低于计算出的清洁限度。杀孢子剂应具有允许的每日暴露量（PDE）或可接受的每日暴露量（ADE）值，并且在计算清洁限度时应考虑共享的表面积。

 

The risks associated with this step include:

 

与这一步骤相关的风险包括：

 

Manual disinfection application and being able to cover the surfaces of the stopper bowl

手动进行消毒操作并确保能够覆盖胶塞的表面

Unidirectional overlapping strokes are challenging to perform due to irregularities on the stopper bowl surface

由于胶塞表面不平整，进行单向重叠擦拭操作具有挑战性

Longer wet-contact times in a Grade A area—The effectiveness of the disinfection is defined as a certain wet-contact time needed to achieve a certain 10-log reduction (7). In a Grade A area, due to the high number of air exchanges, it is challenging to reach wet-contact times longer than five minutes. For Grade A/ISO 5 areas, however, the acceptable microbial count should be 0 (no growth) (2). According to USP 43, Chapter <1116>, samples with contamination should not exceed 0.1% of the total samples collected. In other words, out of 1,000 samples, 999 should show no growth. This implies that, for example, a 10-minute contact time with a sporicidal agent to achieve a 6-log reduction might not be required. A shorter wet-contact time, based on achievable values, can be validated for a Grade A area, reducing the risk of the contact time not being respected. The effectiveness of the procedure can be confirmed during aseptic process simulation and in disinfectant in-situ testing. (See Figure 10.)

在 A 级区域内较长的湿接触时间 —— 消毒的有效性定义为达到一定的对数减少所需的特定湿接触时间（参考文献 7）。在 A 级区域，由于换气次数较多，要达到超过五分钟的湿接触时间具有挑战性。然而，对于 A 级 / ISO 5 区域，可接受的微生物计数应为 0（无生长）（参考文献 2）。根据美国药典（USP）第 43 版第 <1116> 章，受污染的样品不应超过所采集总样品的 0.1%。换句话说，在 1000 个样品中，999 个应无微生物生长。这意味着，例如，为达到 6 个对数减少而使用杀孢子剂接触 10 分钟可能并非必要。对于 A 级区域，可以根据可实现的值验证较短的湿接触时间，从而降低不遵守接触时间的风险。该程序的有效性可以在无菌工艺模拟和消毒剂原位测试中得到确认。（见图 10）

Figure 10 Risk evaluation of the manual disinfection step of a nondetachable stopper bowl

图 10 不可拆卸胶塞手动消毒步骤的风险评估

 

Conclusion

结论

 

The risk-assessment exercise for both scenarios, involving detachable and nondetachable stopper bowls, aimed to identify the major challenges in each case and determine the best recommendations or control measures to mitigate contamination risks.

 

针对可拆卸和不可拆卸胶塞这两种情况所进行的风险评估工作，旨在识别每种情况下的主要挑战，并确定最佳的建议或控制措施，以降低污染风险。

 

It is challenging to reach a conclusion about which scenario poses the lowest risks due to the numerous assumptions made during the analysis. Each manufacturer should arrive at their own conclusions based on their procedures and controls in place. An overview of the advantages, disadvantages and recommendations for both scenarios are given in Table 5.

 

由于在分析过程中做出了大量假设，要得出哪种情况风险最低的结论具有一定挑战性。每个制造商都应根据自身的操作程序和已实施的控制措施得出各自的结论。表 5 给出了这两种情况的优缺点以及建议的概述。

 

Table 5 Summary of advantages, disadvantages and recommendations of both scenarios

表 5 两种情况的优缺点及建议总结

 

The simplest way to comply with EU GMP Annex 1 is to use a detachable stopper bowl, and new filling lines will meet this requirement. Alternatively, redesigning the production line for nondetachable parts is an option, though it may be prohibitively expensive for some companies. Additionally, the more indirect surfaces that cannot be sterilized, the greater the effort and cost required to redesign the filling line. In such cases, a risk-management approach can justify an alternative procedure, such as manual C&D.

 

符合欧盟药品生产质量管理规范（GMP）附件 1 的最简单方法是使用可拆卸的胶塞斗，并且新的灌装线应满足这一要求。或者，对不可拆卸部件的生产线进行重新设计也是一种选择，不过对于某些公司来说，这可能成本过高。此外，无法进行灭菌处理的间接表面越多，重新设计灌装线所需的精力和成本就越大。在这种情况下，采用风险管理方法可以为诸如手动清洁和消毒（C&D）之类的替代程序提供合理依据。

来源：Internet

关键词： 胶塞斗灭菌