你是否了解，新版EU-GMP 附錄 1 已提高了對制藥企業及其供應商的監管要求？讓我們深入探究這對滅菌前的內包裝組件產生何種影響。

 

The Function of Primary Packaging
內包裝的功能

 

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.

內包裝是容器密封系統的組成部分，該系統是所有包裝元件的集合，共同用于容納和保護制劑。容器密封系統還包含外包裝（如紙盒、托盤），這些外包裝可能具備功能性，也可能不具備，且不在本文討論范圍內。

 

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:

內包裝在藥品安全中占據關鍵地位。內包裝組件是指與藥品直接接觸或可能直接接觸的元件，例如：

 

Plastic and aluminum films
塑料和鋁膜

Glass
玻璃

Plastic or elastomer stoppers
塑料或彈性體塞子

Aluminum or plastic tubing
鋁管或塑料管

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

 

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

無菌生產中使用的內包裝組件主要分為兩類：

 

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.

以保護為導向的包裝：這類包裝存在與產品直接接觸的可能性，旨在保護藥品免受外部影響；例如西林瓶和膠塞，它們不具備直接給藥功能，但可維持內容物的完整性。

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.

裝置：這類包裝存在與產品直接接觸的可能性，并積極參與藥物的遞送；典型例子包括用于給藥的注射器和涂藥器。

 

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

這兩種類型的內包裝組件之間的差異對于應用和監管要求均具有重要意義。

 

Regulatory Guidelines and Their Implications
監管指南及其影響

 

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).

根據《美國藥典》的闡述，內包裝可能成為微生物污染的來源，其初始質量、儲存條件、制備和處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 附錄 1 規定，無菌產品的生產必須遵循特殊要求，以最大限度降低微生物、顆粒和內毒素 / 熱原污染的風險。第 2.1 節明確 “應考慮以下關鍵領域：iv. 應對包裝材料進行全面控制和測試，以確保其生物負載和內毒素 / 熱原水平適合使用”（2）。此外，根據 GMP 附錄1 的原則，應實施監測系統以證明已實施基于風險的控制措施，且符合質量預期。

 

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.

明確要求滅菌工藝（包括采用過度殺滅方法的工藝）應進行生物負載監測（3）。這種監測對于滿足監管要求和質量標準（如美國食品藥品監督管理局（FDA）、歐洲藥品管理局（EMA）和國際標準化組織（ISO）制定的標準）不可或缺，并確保在整個生命周期內維持無菌狀態。

 

Understanding Bioburden and Its Role
了解生物負載及其作用

 

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??). Achieving this sterility assurance level relies on two fundamental pillars:

生物負載是指與特定物品（如內包裝組件）相關的微生物總數。生物負載是無菌產品生產中的決定性因素。在此背景下，無菌性意味著滅菌后存在存活微生物的概率不超過百萬分之一（10??）。實現這一無菌保證水平依賴于兩大基本支柱：

 

Control and continuous monitoring of hygiene conditions throughout the manufacturing process
整個生產過程中衛生條件的控制和持續監測

The application of a scientifically validated sterilization process
使用經過科學驗證的滅菌工藝

 

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.

生物負載被視為一種威脅，不僅源于滅菌工藝可能失敗，還因為可能存在過敏原、內毒素和外毒素等殘留物質。

 

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.

關于內毒素，滅菌工藝本身存在一定風險。內毒素可耐受滅菌工藝，若殘留在藥品中可能引發健康并發癥。這凸顯了控制包括內包裝在內的內毒素輸入水平的重要性。滅菌工藝的驗證階段和批次放行過程均需進行內毒素檢測，以檢測和量化這些物質，確保藥品安全。

 

Risk Assessment
風險評估

 

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，制造商必須制定并考慮無菌生產過程中所有步驟的策略和控制措施，包括進廠內包裝組件。

 

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.
風險評估是 CCS 的基礎。因此，第一步是進行無菌保證風險評估，以確定生物負載控制措施和關鍵控制點。

 

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.
風險評估范圍從供應商資質審核延伸至包裝滅菌環節（最終滅菌或干熱滅菌后無菌灌裝）。供應商可提供未處理組件和即用滅菌（RTS）組件。與 RTS 材料相比，未處理組件通常缺乏供應商提供的生物負載數據。對于這些未處理組件，供應商需進行符合風險評估要求的專門清潔或控制。

 

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).
應在潛在生物負載降低工藝前后以及滅菌前的工藝節點設置監測點，以確保最終滅菌步驟能有效降低生物負載，或確保無菌灌裝從無污染的批量物料開始（1,4）。

 

During the supplier qualification, suppliers should demonstrate that starting-packaging components meet established criteria and purchasing specifications (5).
在供應商資質審核過程中，供應商應證明內包裝組件符合既定標準和采購規格（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).
制定最終滅菌或去熱原前的清洗工藝，以確保充分降低進廠物料的污染。在制定清潔驗證方案時，應考慮微生物和內毒素污染的風險（6）。

 

The Importance of a Testing Program
檢測程序的重要性

 

The primary purposes of maintaining a testing program are to:
維護檢測程序的主要目的在于：

 

Ensure the microbial load remains acceptable throughout the process

確保整個過程中的微生物負載保持在可接受范圍內
Assure the item meets the required acceptable limit for bioburden

確保物品符合生物負載的可接受限值要求
Evaluate the effectiveness of all controls

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

管理藥品質量和安全風險

 

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).
該程序應提供趨勢分析數據，以評估控制措施的整體有效性，確保符合 GMP 要求，發現偏差并推動持續改進。程序應包括有記錄且合理的采樣點、樣本量、采樣頻率和檢測方法。樣本數量需通過統計確定，并在采樣計劃中明確規定（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.
監測頻率需根據具體情況確定，取決于滅菌工藝、歷史數據、污染風險和放行流程。

 

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.
監測不足會導致污染未被發現，或錯過超出趨勢的早期信號和即將到來的污染風險。相反，過度監測可能帶來負擔、缺乏實際信息且成本高昂，并可能在工藝中引入污染（8）。此外，假陽性風險增加，導致不必要的調查和產品放行延遲。

 

For components with a potential to introduce contamination, each lot must to be tested prior to use (9).
對于可能引入污染的組件，每批使用前必須進行檢測（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.
對于最終滅菌產品（2），如果使用過度殺滅參數且滅菌前進行了驗證的清潔，則可定期進行監測。

 

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).
對于經認證的包裝，采購規格和檢測結果在分析證書中進行控制。然而，全球監管機構要求藥品制造商定期進行確認檢測，以驗證供應商分析證書中提供的結果（9）。

 

Identification: Know Your Enemy
鑒定：了解你的 “敵人”

 

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.
通過詳細準確的鑒定程序，可以觀察到微生物菌群的變化或特定微生物的優勢地位。這有助于在偏差期間確定污染源，并采取適當的預防措施以降低未來污染的風險。

 

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).
對于警戒限和行動限偏差以及參數放行，需對生物負載檢測中發現的任何微生物進行鑒定和評估。需考慮所回收微生物的特性（如有毒副產物），以確定其對產品質量和滅菌工藝有效性的影響。與藥品生物負載偏差類似，需適當監測內毒素 / 熱原水平（2）。

 

Setting Appropriate Limits
設定適當的限度

 

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).
為未處理組件和即用滅菌（RTS）組件設定適當的接受限度至關重要。設定限度是為了確保產品質量，并與滅菌前生物負載降低工藝的能力一致。然而，不得通過滅菌工藝或滅菌前任何生物負載降低步驟的能力來證明高限度的合理性（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.
針對特定滅菌工藝的驗證研究中需確定適當的行動限。這些研究包括對包裝進行采樣和檢測，以量化微生物負載，并評估滅菌工藝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 方法進行伽馬輻照時，生物負載行動限的最大值為 1000 CFU / 產品單元（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).
經過驗證的干熱（>220°C）通常用于耐熱容器組件的滅菌和去熱原。這些去熱原循環屬于過度殺滅循環，已證明可使耐熱內毒素減少 3 個對數單位（10）。這可能無需進行生物負載監測，僅內毒素監測即可（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).
歐洲藥品管理局（EMA）關于內包裝容器滅菌的指南建議，蒸汽滅菌循環（包括過度殺滅循環）前的生物負載水平為 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).
過度殺滅循環并不總是適用于有熱標記的產品或物品。在這種情況下，滅菌循環需基于所回收生物負載微生物的耐熱性和數量進行設計。循環開發過程中需包含安全裕度，該裕度與生物負載監測程序的頻率和強度呈負相關（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.
除干熱、蒸汽滅菌和輻照外，還有氣體滅菌（如環氧乙烷滅菌）。同樣，氣體滅菌工藝前的生物負載應降至最低，并按照 ISO 標準和引用的 EMA 指令設定限度。

 

Alert limits are used for trending purposes and based on historical data.
警戒限用于趨勢分析，基于歷史數據設定。

 

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《歐洲藥典》第 2.6.12 章所述，若將組件混合作為樣品制備以進行計數測試，限度應以 CFU / 測試組件數量表示。

 

The Need for Regular Data Review
定期數據審查的必要性

 

Periodic data review is required to evaluate the state of control and to identify the occurrence of adverse trends. An adverse trend includes:
需定期進行數據審查，以評估控制狀態并識別不良趨勢的發生。不良趨勢包括：

 

Increasing number of excursions from action limits or alert levels

超出行動限或警戒限的次數增加
Successive excursions from alert levels

連續超出警戒限
Regular isolated excursions from action limits

定期出現孤立的超出行動限情況
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)
微生物菌群類型和數量的變化以及特定微生物的優勢地位（應注意可能表明失控的回收微生物，如產芽孢微生物）

 

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.
出現不良趨勢時，需進行調整和改進，并通過調查確定是否存在失控及影響產品質量的風險。

 

Conclusion
結論

 

Bioburden monitoring of primary-packaging components is crucial for ensuring product safety and quality.
內包裝組件的生物負載監測對確保產品安全和質量至關重要。

 

A risk-based approach is essential, as it enables customized control strategies that address specific risks associated with different process steps and sterilization methods.
基于風險的方法至關重要，因為它能夠制定定制化的控制策略，以應對與不同工藝步驟和滅菌方法相關的特定風險。

 

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.
生物負載限度不應僅依賴滅菌工藝消除污染物的能力。相反，應實施全面的控制策略，包括對生物負載數據的定期審查。審查旨在評估控制狀態并識別任何不良趨勢，從而確保持續改進并維持高標準的產品安全。