你是否了解，新版EU-GMP 附录 1 已提高了对制药企业及其供应商的监管要求？让我们深入探究这对灭菌前的内包装组件产生何种影响。

 

The Function of Primary Packaging
内包装的功能

 

Primary packaging serves as a component of the container–closure system, which represents the aggregate of packaging elements that collectively contain and safeguard the dosage form. The container–closure system additionally encompasses secondary packaging (e.g., cartons, trays), which may or may not possess functional attributes and fall outside the scope of this discussion.

内包装是容器密封系统的组成部分，该系统是所有包装元件的集合，共同用于容纳和保护制剂。容器密封系统还包含外包装（如纸盒、托盘），这些外包装可能具备功能性，也可能不具备，且不在本文讨论范围内。

 

Primary packaging holds a pivotal role in drug safety. Primary-packaging components refer to elements that are in direct contact with, or have the potential to come into direct contact with, the drug product, such as:

内包装在药品安全中占据关键地位。内包装组件是指与药品直接接触或可能直接接触的元件，例如：

 

Plastic and aluminum films
塑料和铝膜

Glass
玻璃

Plastic or elastomer stoppers
塑料或弹性体塞子

Aluminum or plastic tubing
铝管或塑料管

Containers (and their closures)
容器（及其密封件）

 

Primary-packaging components utilized in sterile manufacturing are categorized into two main types:

无菌生产中使用的内包装组件主要分为两类：

 

Protection-oriented packaging: These involve potential direct product contact and are engineered to shield the drug from external influences; instances include vials and stoppers, which lack direct administration functionality but preserve the integrity of the contents.

以保护为导向的包装：这类包装存在与产品直接接触的可能性，旨在保护药品免受外部影响；例如西林瓶和胶塞，它们不具备直接给药功能，但可维持内容物的完整性。

Devices: These entail potential direct product contact and actively participate in the delivery of the medication; typical instances are syringes and applicators employed for drug administration.

装置：这类包装存在与产品直接接触的可能性，并积极参与药物的递送；典型例子包括用于给药的注射器和涂药器。

 

The differentiation between these two types of primary-packaging components is significant for both application and regulatory requirements.

这两种类型的内包装组件之间的差异对于应用和监管要求均具有重要意义。

 

Regulatory Guidelines and Their Implications
监管指南及其影响

 

As outlined in the U.S. Pharmacopeia, primary packaging can act as a source of microbial contamination, and their initial quality, storage conditions, preparation, and handling procedures must be taken into account (1).

根据《美国药典》的阐述，内包装可能成为微生物污染的来源，其初始质量、储存条件、制备和处理程序均需予以考量（1）。

 

GMP Annex 1 stipulates that the manufacture of sterile products must adhere to special requirements to minimize the risk of microbial, particulate, and endotoxin/pyrogen contamination. Section 2.1 specifies that “the following key areas should be addressed: iv. Packaging materials should be thoroughly controlled and tested to ensure that bioburden and endotoxin/pyrogen levels are appropriate for use” (2). Furthermore, in accordance with the principles of GMP Annex 1, monitoring systems should be implemented to demonstrate that risk-based controls are being applied and quality expectations are being fulfilled.

GMP 附录 1 规定，无菌产品的生产必须遵循特殊要求，以最大限度降低微生物、颗粒和内毒素 / 热原污染的风险。第 2.1 节明确 “应考虑以下关键领域：iv. 应对包装材料进行全面控制和测试，以确保其生物负载和内毒素 / 热原水平适合使用”（2）。此外，根据 GMP 附录1 的原则，应实施监测系统以证明已实施基于风险的控制措施，且符合质量预期。

 

It is explicitly required that sterilization processes (including those utilizing overkill methods) be supported by a bioburden-monitoring program (3). Monitoring is indispensable for meeting regulatory requirements and quality standards, such as those established by the U.S. Food and Drug Administration (FDA), European Medicines Agency (EMA), and International Organization for Standardization (ISO), and for ensuring sterility is maintained throughout the lifecycle.

明确要求灭菌工艺（包括采用过度杀灭方法的工艺）应进行生物负载监测（3）。这种监测对于满足监管要求和质量标准（如美国食品药品监督管理局（FDA）、欧洲药品管理局（EMA）和国际标准化组织（ISO）制定的标准）不可或缺，并确保在整个生命周期内维持无菌状态。

 

Understanding Bioburden and Its Role
了解生物负载及其作用

 

Bioburden refers to the total number of microorganisms associated with a specific item, such as primary-packaging components. Bioburden is a determining factor in the manufacture of sterile products. In this context, sterility implies that the probability of viable microorganisms existing after sterilization does not exceed one in one million (10⁻⁶). Achieving this sterility assurance level relies on two fundamental pillars:

生物负载是指与特定物品（如内包装组件）相关的微生物总数。生物负载是无菌产品生产中的决定性因素。在此背景下，无菌性意味着灭菌后存在存活微生物的概率不超过百万分之一（10⁻⁶）。实现这一无菌保证水平依赖于两大基本支柱：

 

Control and continuous monitoring of hygiene conditions throughout the manufacturing process
整个生产过程中卫生条件的控制和持续监测

The application of a scientifically validated sterilization process
使用经过科学验证的灭菌工艺

 

Bioburden is regarded as a threat not only because of the potential for unsuccessful sterilization processes but also due to the possible presence of residual substances such as allergens, endotoxins, and exotoxins.

生物负载被视为一种威胁，不仅源于灭菌工艺可能失败，还因为可能存在过敏原、内毒素和外毒素等残留物质。

 

Regarding endotoxins, the sterilization process itself presents certain risks. Endotoxins can withstand sterilization processes and cause health complications if they remain in the drug. This underscores the importance of controlling incoming endotoxin levels, including those from primary packaging. Both the validation phase of the sterilization process and the batch release process require endotoxin testing to detect and quantify these substances, ensuring drug safety.

关于内毒素，灭菌工艺本身存在一定风险。内毒素可耐受灭菌工艺，若残留在药品中可能引发健康并发症。这凸显了控制包括内包装在内的内毒素输入水平的重要性。灭菌工艺的验证阶段和批次放行过程均需进行内毒素检测，以检测和量化这些物质，确保药品安全。

 

Risk Assessment
风险评估

 

In the contamination control strategy (CCS), manufacturers must state and consider their own strategies and controls for all steps in the sterile manufacturing process, including the incoming primary-packaging components.

在污染控制策略（CCS）中，制造商必须制定并考虑无菌生产过程中所有步骤的策略和控制措施，包括进厂内包装组件。

 

Risk assessments are the foundation of the CCS. Therefore, the first step is a sterility-assurance risk assessment to define bioburden control measures and critical control points.
风险评估是 CCS 的基础。因此，第一步是进行无菌保证风险评估，以确定生物负载控制措施和关键控制点。

 

Risk assessment extends from the supplier qualification to the point where the packaging is sterilized (either terminal or dry-heat sterilized and then aseptically filled). Suppliers can provide untreated components and ready-to-sterilize (RTS) components. Less bioburden data from the supplier is often known for untreated components, in contrast to RTS materials. For these untreated components, the supplier needs specialized cleaning or controls that meet the risk assessment requirements.
风险评估范围从供应商资质审核延伸至包装灭菌环节（最终灭菌或干热灭菌后无菌灌装）。供应商可提供未处理组件和即用灭菌（RTS）组件。与 RTS 材料相比，未处理组件通常缺乏供应商提供的生物负载数据。对于这些未处理组件，供应商需进行符合风险评估要求的专门清洁或控制。

 

Monitoring points should be considered at the points in the process either immediately before or after a potential bioburden-reduction process and immediately prior to sterilization in order to provide confidence that the terminal sterilization step will effectively reduce the bioburden or that aseptic filling begins with an untainted bulk (1, 4).
应在潜在生物负载降低工艺前后以及灭菌前的工艺节点设置监测点，以确保最终灭菌步骤能有效降低生物负载，或确保无菌灌装从无污染的批量物料开始（1,4）。

 

During the supplier qualification, suppliers should demonstrate that starting-packaging components meet established criteria and purchasing specifications (5).
在供应商资质审核过程中，供应商应证明内包装组件符合既定标准和采购规格（5）。

 

Washing processes prior to terminal sterilization or depyrogenation are developed to ensure adequate reduction of incoming contamination. The risk of microbial and endotoxin contamination should be considered in the development of cleaning validation protocols (6).
制定最终灭菌或去热原前的清洗工艺，以确保充分降低进厂物料的污染。在制定清洁验证方案时，应考虑微生物和内毒素污染的风险（6）。

 

The Importance of a Testing Program
检测程序的重要性

 

The primary purposes of maintaining a testing program are to:
维护检测程序的主要目的在于：

 

Ensure the microbial load remains acceptable throughout the process

确保整个过程中的微生物负载保持在可接受范围内
Assure the item meets the required acceptable limit for bioburden

确保物品符合生物负载的可接受限值要求
Evaluate the effectiveness of all controls

评估所有控制措施的有效性
Manage risks to drug product quality and safety

管理药品质量和安全风险

 

The program should provide data for trending to evaluate the collective effectiveness of control measures to ensure GMP compliance, detect deviations and drive continuous improvement. A program will include documented and justified sampling points, sample size and frequency, and test methods. The number of samples is statistically determined and specified in a sampling plan (7).
该程序应提供趋势分析数据，以评估控制措施的整体有效性，确保符合 GMP 要求，发现偏差并推动持续改进。程序应包括有记录且合理的采样点、样本量、采样频率和检测方法。样本数量需通过统计确定，并在采样计划中明确规定（7）。

 

The monitoring frequency is determined on a case-by-case basis. It depends on the sterilization process, historical data, contamination risk and the release process.
监测频率需根据具体情况确定，取决于灭菌工艺、历史数据、污染风险和放行流程。

 

Inadequate monitoring results in undetected contamination or in early signals of out-of-trends and impending contamination risks being missed. Conversely, excessive monitoring can be burdensome, uninformative and costly and may introduce contamination to the process (8). Additionally, the risk of false positives increases, resulting in unneeded investigations and delays of product release.
监测不足会导致污染未被发现，或错过超出趋势的早期信号和即将到来的污染风险。相反，过度监测可能带来负担、缺乏实际信息且成本高昂，并可能在工艺中引入污染（8）。此外，假阳性风险增加，导致不必要的调查和产品放行延迟。

 

For components with a potential to introduce contamination, each lot must to be tested prior to use (9).
对于可能引入污染的组件，每批使用前必须进行检测（9）。

 

In line with terminally sterilized products (2), monitoring at intervals is possible if overkill parameters are used and a validated cleaning is performed prior to sterilization.
对于最终灭菌产品（2），如果使用过度杀灭参数且灭菌前进行了验证的清洁，则可定期进行监测。

 

For certified packaging, the purchasing specifications and test results are controlled on the Certificate of Analysis. However, global regulatory authorities expect the drug product manufacturer to perform confirmation testing periodically to verify the results provided on the supplier Certificate of Analysis (9).
对于经认证的包装，采购规格和检测结果在分析证书中进行控制。然而，全球监管机构要求药品制造商定期进行确认检测，以验证供应商分析证书中提供的结果（9）。

 

Identification: Know Your Enemy
鉴定：了解你的 “敌人”

 

Through a detailed and accurate identification program, shifts in the microbial flora or the predominance of specific organisms are observed. It helps to identify the source of contamination during excursions and to take appropriate preventive measures to reduce the risk of future contamination.
通过详细准确的鉴定程序，可以观察到微生物菌群的变化或特定微生物的优势地位。这有助于在偏差期间确定污染源，并采取适当的预防措施以降低未来污染的风险。

 

For alert and action-limit excursions and at parametric release, any organisms found during bioburden testing are identified and evaluated. The characteristics (e.g., by-products as toxins) of the recovered organism are considered to determine the impact on product quality and the effectiveness of the sterilization process. Similar to drug-product bioburden excursions, endotoxin/pyrogen levels are monitored as appropriate (2).
对于警戒限和行动限偏差以及参数放行，需对生物负载检测中发现的任何微生物进行鉴定和评估。需考虑所回收微生物的特性（如有毒副产物），以确定其对产品质量和灭菌工艺有效性的影响。与药品生物负载偏差类似，需适当监测内毒素 / 热原水平（2）。

 

Setting Appropriate Limits
设定适当的限度

 

Establishing appropriate acceptance limits for packaging components is important for untreated and RTS components. Limits are set to ensure the product quality and to be consistent with presterilization bioburden reduction process capability. However, high limits should not be justified by the capacity of the sterilization process or any bioburden-reduction step prior to sterilization (10).
为未处理组件和即用灭菌（RTS）组件设定适当的接受限度至关重要。设定限度是为了确保产品质量，并与灭菌前生物负载降低工艺的能力一致。然而，不得通过灭菌工艺或灭菌前任何生物负载降低步骤的能力来证明高限度的合理性（10）。

 

Appropriate action limits are determined during validation studies for the specific sterilization process. These studies include sampling and testing of packaging to quantify the microbial load and evaluate the effectiveness of the sterilization process and control measures.
针对特定灭菌工艺的验证研究中需确定适当的行动限。这些研究包括对包装进行采样和检测，以量化微生物负载，并评估灭菌工艺和控制措施的有效性。

 

Gamma irradiation using the ISO 11137 VDmax25 method has a maximum bioburden action limit of 1,000 CFU/product unit (11).
使用 ISO 11137 VDmax25 方法进行伽马辐照时，生物负载行动限的最大值为 1000 CFU / 产品单元（11）。

 

Dry heat (at >220 °C) for a validated time is often used for both sterilization and depyrogenation of heat-resistant container components. These depyrogenation cycles are overkill cycles and demonstrated a 3-log reduction in heat-resistant endotoxins (10). It may eliminate the need for the bioburden monitoring, and endotoxin monitoring is sufficient (3).
经过验证的干热（>220°C）通常用于耐热容器组件的灭菌和去热原。这些去热原循环属于过度杀灭循环，已证明可使耐热内毒素减少 3 个对数单位（10）。这可能无需进行生物负载监测，仅内毒素监测即可（3）。

 

The EMA guideline for sterilization of primary containers recommends a bioburden level of 100 CFU (per 100mL) prior to steam sterilization cycles (including overkill cycle) (10).
欧洲药品管理局（EMA）关于内包装容器灭菌的指南建议，蒸汽灭菌循环（包括过度杀灭循环）前的生物负载水平为 100 CFU（每 100mL）（10）。

 

Overkill cycles cannot always be applied for heat-labeled products or items. In that case, sterilization cycles are designed based on heat resistance and population of the bioburden recovered organisms. A safety margin during the cycle development is included, inversely correlated to the frequency and magnitude of the bioburden-monitoring program (12).
过度杀灭循环并不总是适用于有热标记的产品或物品。在这种情况下，灭菌循环需基于所回收生物负载微生物的耐热性和数量进行设计。循环开发过程中需包含安全裕度，该裕度与生物负载监测程序的频率和强度呈负相关（12）。

 

In addition to dry heat, steam sterilization and irradiation, there is also gas sterilization, for example, with ethylene oxide. Once again, bioburden prior to the gas sterilization process should be minimized, and limits are established, as described by ISO standards and the referenced EMA directive.
除干热、蒸汽灭菌和辐照外，还有气体灭菌（如环氧乙烷灭菌）。同样，气体灭菌工艺前的生物负载应降至最低，并按照 ISO 标准和引用的 EMA 指令设定限度。

 

Alert limits are used for trending purposes and based on historical data.
警戒限用于趋势分析，基于历史数据设定。

 

If components are mixed as a sample preparation to perform the enumeration test, as described in chapter 2.6.12 of the European Pharmacopoeia, the limit should be expressed as CFU per number of components tested.
如《欧洲药典》第 2.6.12 章所述，若将组件混合作为样品制备以进行计数测试，限度应以 CFU / 测试组件数量表示。

 

The Need for Regular Data Review
定期数据审查的必要性

 

Periodic data review is required to evaluate the state of control and to identify the occurrence of adverse trends. An adverse trend includes:
需定期进行数据审查，以评估控制状态并识别不良趋势的发生。不良趋势包括：

 

Increasing number of excursions from action limits or alert levels

超出行动限或警戒限的次数增加
Successive excursions from alert levels

连续超出警戒限
Regular isolated excursions from action limits

定期出现孤立的超出行动限情况
Changes in the type and number of microbial flora and the predominance of specific organisms (attention should be paid to recovered organisms that may indicate a loss of control, e.g., spore-forming microorganisms)
微生物菌群类型和数量的变化以及特定微生物的优势地位（应注意可能表明失控的回收微生物，如产芽孢微生物）

 

At adverse trends, adjustments and improvements are made, and the investigation determines if there is a loss of control and risk of impacting product quality.
出现不良趋势时，需进行调整和改进，并通过调查确定是否存在失控及影响产品质量的风险。

 

Conclusion
结论

 

Bioburden monitoring of primary-packaging components is crucial for ensuring product safety and quality.
内包装组件的生物负载监测对确保产品安全和质量至关重要。

 

A risk-based approach is essential, as it enables customized control strategies that address specific risks associated with different process steps and sterilization methods.
基于风险的方法至关重要，因为它能够制定定制化的控制策略，以应对与不同工艺步骤和灭菌方法相关的特定风险。

 

Bioburden limits should not rely solely on the sterilization process’s capacity to eliminate contaminants. Instead, a comprehensive control strategy is implemented, incorporating periodic reviews of bioburden data. Reviews are for evaluating the state of control and identifying any adverse trends, thereby ensuring continuous improvement and maintaining high standards of product safety.
生物负载限度不应仅依赖灭菌工艺消除污染物的能力。相反，应实施全面的控制策略，包括对生物负载数据的定期审查。审查旨在评估控制状态并识别任何不良趋势，从而确保持续改进并维持高标准的产品安全。