stoppering on line; another stage included filling 3000 vials, transportation to the lyophilizer and then stoppering; a third stage included the filling of 3000 vials, loading in the lyophilizer, and exposure to a portion of the nitrogen flush and then stoppering. Since lyophilizer sterilization and sterilization of the nitrogen system used to backfill require separate validation, media fills should primarily validate the filling, transportation and loading aseptic operations.
为了努力确定灌装和无菌操作的单元操作可能带入的污染，一些生产商采取增加培养基灌装量。他们在培养基灌装中大约灌装9000瓶，把灌装过程分割为3个阶段。第一部分是灌装3000瓶并且加塞，第二部分是灌装3000瓶，传递到冻干机中再全加塞，第三部分是灌装3000瓶，排放到冻干机内，暴露在氮气流下冻干机的灭菌和用于回填的无菌氮气系统分别进行验证，培养基灌装主要验证灌装、传递和摆放的无菌操作。
The question of the number of units needed for media fills when the capacity of the process is less than 3000 units is frequently asked, particularly for clinical products. Again, the purpose of the media fill is to assure that product can be aseptically processed without contamination under operating conditions. It would seem, therefore, that the maximum number of units of media filled be equivalent to the maximum batch size if it is less than 3000 units.
有个问题经常被问到，就是当生产能力小于3000瓶时的培养基灌装数量，特别是在进行临床样品制备时。培养基灌装的目的是保证在操作条件下的无菌过程不被污染物污染。因此当最大的批产量小于3000瓶时，培养基灌装的数量应等同于最大的批产量。
After filling, dosage units are transported to the lyophilizer by metal trays. Usually, the bottom of the trays are removed after the dosage units are loaded into the lyophilizer. Thus, the dosage units lie directly on the lyophilizer shelf. There have been some situations in which manufacturers have loaded the dosage units on metal trays which were not removed. Unfortunately, at one manufacturer, the trays warped which caused a moisture problem in some dosage units in a batch.
灌装后药物通过金属托盘传递到冻干机里。通常当药物摆放到冻干机里后，托盘底部要被抽走。从而药物直接摆放在冻干机的板层上。有些情况是，生产商摆放药物时不抽走托盘底部。不幸的是托盘的不平会导致一批内的一些产品潮湿问题。
In the transport of vials to the lyophilizer, since they are not sealed, there is concern for the potential for contamination. During inspections and in the review of new facilities, the failure to provide laminar flow coverage or a primary barrier for the transport and loading areas of a lyophilizer has been regarded as an objectionable condition. One manufacturer as a means of correction developed a laminar flow cart
to transport the vials from the filling line to the lyophilizer. Other manufacturers building new facilities have located the filling line close to the lyophilizer and have provided a primary barrier extending from the filling line to the lyophilizer.
在传递瓶子到冻干机里，由于瓶子不是密闭的，所以存在潜在的污染可能性。在检查和回顾一些新的工厂时，在传递和摆放到冻干机的区域缺乏一个层流罩或主要的屏障被认为是不可接收的。有的生产商的补救措施是使用层流车，在灌装线和冻干机之间传递瓶子。还有的生产商建造新的车间时，将灌装线和冻干机摆放的很近，并且在灌装机和冻干机之间安装主要的屏障。
In order to correct this type of problem, another manufacturer installed a vertical laminar flow hood between the filling line and lyophilizer. Initially, high velocities with inadequate return caused a contamination problem in a media fill. It was speculated that new air currents resulted in rebound contamination off the floor. Fortunately, media fills and smoke studies provided enough meaningful information that the problem could be corrected prior to the manufacture of product. Typically, the lyophilization process includes the stoppering of vials in the chamber.
为了解决这类问题，一些生产商在灌装线和冻干机之间安装层流罩。最初由于周转频率太快，导致了培养基灌装的污染问题。推测可能是气流由地板反弹造成的污染。幸运的是培养基灌装和烟笔试验提供了足够的有用信息，能够在生产产品前纠正问题。冻干过程包括在冻干机前箱内的全加塞。
Another major concern with the filling operation is assurance of fill volumes. Obviously, a low fill would represent a subpotency in the vial. Unlike a powder or liquid fill, a low fill would not be readily apparent after lyophilization particularly for a biopharmaceutical drug product where the active ingredient may be only a milligram. Because of the clinical significance, sub-potency in a vial potentially can be a very serious situation.
另一个主要的问题是灌装过程中保证灌装量。很明显低灌装量会造成药物效力的降低。不象粉末和液体灌装，冻干后的产品不能明显的发现低灌装量。特别是生物药品，它们的活性成分可能仅有几毫克。因为临床的重要性，潜在的药效降低将被严肃对待。
For example, in the inspection of a lyophilization filling operation, it was noted that the firm was having a filling problem. The gate on the filling line was not coordinated with the filling syringes, and splashing and partial filling was occurring. It was also observed that some of the partially filled vials were loaded into the lyophilizer. This resulted in rejection of the batch.
例如，检查一个冻干的灌装操作，显示了这个公司存在灌装操作问题。灌装线上的闸门没有按照灌装注射器进行调整，洒料，不平均的灌装是事故。观察到一些
部分灌装的产品被装载到冻干机。这样的结果是拒绝这批产品。
On occasion, it has been seen that production operators monitoring fill volumes record these fill volumes only after adjustments are made. Therefore, good practice and a good quality assurance program would include the frequent monitoring of the volume of fill, such as every 15 minutes. Good practice would also include provisions for the isolation of particular sections of filling operations when low or high fills are encountered.
观察灌装量只是通过检查操作人的灌装量监控记录，灌装量是通过调整调节器决定的。好的方法和好的质量管理程序是包括装量检查的频率，例如每15分钟检查一次。好的方法是当发现低或高灌装量时应隔离。
There are some atypical filling operations which have not been discussed. For example, there have also been some situations in which lyophilization is performed on trays of solution rather than in vials. Based on the current technology available, it would seem that for a sterile product, it would be difficult to justify this procedure. 一些非典型的灌装操作没有讨论，例如，有些冻干是在托盘上进行的而不是在瓶子里，基于现在的技术条件，把它看成无菌产品，是很难被证明是正当的。 The dual chamber vial also presents additional requirements for aseptic manipulations. Media fills should include the filling of media in both chambers. Also, the diluent in these vials should contain a preservative. (Without a preservative, the filling of diluent would be analogous to the filling of media. In such cases, a 0% level of contamination would be expected.)
双腔体瓶子对无菌操作提出了新的要求。培养基灌装必须包括灌装两个腔体。 LYOPHILIZATION CYCLE AND CONTROLS 冻干周期控制
After sterilization of the lyophilizer and aseptic loading, the initial step is freezing the solution. In some cycles, the shelves are at the temperature needed for freezing, while for other cycles, the product is loaded and then the shelves are taken to the freezing temperature necessary for product freeze. In those cycles in which the shelves are precooled prior to loading, there is concern for any ice formation on shelves prior to loading. Ice on shelves prior to loading can cause partial or complete stoppering of vials prior to lyophilization of the product. A recent field complaint of a product in solution and not lyophilized was attributed to preliminary stoppering of a few vials prior to exposure to the lyophilization cycle. Unfortunately, the firm's 100% vial inspection failed to identify the defective vial.
冻干机灭菌和无菌摆放后的第一个步骤是冷冻溶液。在一些冻干周期里板层已经冷冻到冷冻温度，在另外一些循环中，当产品摆放到冻干机板层后才开始进行冷冻到冷冻温度。在预先冷冻的循环过程中，在板层表面在产品摆放之前会结冰。在摆放产品前板层结冰，会导致冻干开始前瓶子被部分或完全封闭。最近的一些
投诉，表明一些预先被封闭的瓶子没有经过冻干的步骤，造成了产品处于液体状态或没有冻干。不幸的是，这些公司100%的瓶子检查没有将有缺陷的瓶子鉴别出来。
Typically, the product is frozen at a temperature well below the eutectic point. 产品的冷冻温度应低于它的共晶点。
The scale-up and change of lyophilization cycles, including the freezing procedures, have presented some problems. Studies have shown the rate and manner of freezing may affect the quality of the lyophilized product. For example, slow freezing leads to the formation of larger ice crystals. This results in relatively large voids, which aid in the escape of water vapor during sublimation. On the other hand, slow freezing can increase concentration shifts of components. Also, the rate and manner of freezing has been shown to have an affect on the physical form (polymorph) of the drug substance. 放大的和变更的冻干循环，包括冷冻程序，表现出一些问题。研究表明冷冻的速度和方式影响冻干产品的质量。例如，缓慢冷冻造成较大的冰晶排列，造成冰晶升华时有较大的逃逸通道。另一方面缓慢冷冻会造成产品组分的浓度上升。冷冻的速度和方式会影响药物组分的物理形态（同质异构）。
It is desirable after freezing and during primary drying to hold the drying temperature (in the product) at least 4-5o below the eutectic point. Obviously, the manufacturer should know the eutectic point and have the necessary instrumentation to assure the uniformity of product temperatures. The lyophilizer should also have the necessary instrumentation to control and record the key process parameters. These include: shelf temperature, product temperature, condenser temperature, chamber pressure and condenser pressure. The manufacturing directions should provide for time, temperature and pressure limits necessary for a lyophilization cycle for a product. The monitoring of product temperature is particularly important for those cycles for which there are atypical operating procedures, such as power failures or equipment breakdown.
冷冻以后，在主干燥阶段，控制产品的干燥温度低于共晶点4-5℃。显然生产商需要知道产品的共晶点，而且需要使用仪器保证产品的温度是均一的。冻干机要有必须的仪器控制和记录关键的过程参数。包括：板层温度、产品温度、冷凝器温度、前箱压力、冷凝器压力。生产说明中应注明冻干循环中的时间压力和温度限制。在一些非典型的操作程序中（例如断电或设备故障）监测产品的温度是十分重要的。
Electromechanical control of a lyophilization cycle has utilized cam-type recorder-controllers. However, newer units provide for microcomputer control of the freeze drying process. A very basic requirement for a computer controlled process is a flow chart or logic. Typically, operator involvement in a computer controlled lyophilization cycle primarily occurs at the beginning. It consists of loading the
chamber, inserting temperature probes in product vials, and entering cycle parameters such as shelf temperature for freezing, product freeze temperature, freezing soak time, primary drying shelf temperature and cabinet pressure, product temperature for establishment of fill vacuum, secondary drying shelf temperature, and secondary drying time.
电子控制冻干循环一般使用多通道记录仪。一些新的设备配备了电脑，用于控制冻干过程。对电脑控制过程的最基本的要求是流程图和逻辑控制。包括前箱的摆放、向产品瓶中插入温度探头，输入循环参数，例如：板层冷冻温度、产品冷冻温度、追加冷冻时间、主干燥时的板层温度和前箱压力、开始抽真空时的产品温度、第二次干燥时的板层温度、第二次干燥的时间。
In some cases, manufacturers have had to continuously make adjustments in cycles as they were being run. In these situations, the lyophilization process was found to be non-validated.
在一些案例中，生产商在循环开始前不断的调整参数。在这种情况下，冻干过程被认为是没有被经过验证的。
Validation of the software program of a lyophilizer follows the same criteria as that for other processes. Basic concerns include software development, modifications and security. The Guide to Inspection of Computerized Systems in Drug Processing contains a discussion on potential problem areas relating to computer systems. A Guide to the Inspection of Software Development Activities is a reference that provides a more detailed review of software requirements.
冻干机的软件系统验证和其他过程的验证相同。包括软件的变革、修改和安全。《Guide to Inspection of Computerized Systems in Drug Processing》讨论了相关的问题。《Guide to the Inspection of Software Development Activities》提供了逐条的软件需求。
Leakage into a lyophilizer may originate from various sources. As in any vacuum chamber, leakage can occur from the atmosphere into the vessel itself. Other sources are media employed within the system to perform the lyophilizing task. These would be the thermal fluid circulated through the shelves for product heating and cooling, the refrigerant employed inside the vapor condenser cooling surface and oil vapors that may migrate back from the vacuum pumping system.
冻干机的泄漏有不同的来源。象一些真空箱，泄漏使空气进入容器内部。其他来源是冻干过程使用的一些媒介。包括为产品加热和制冷的热传递液体；冷凝蒸发器内制冷剂；真空泵油的回流。
Any one, or a combination of all, can contribute to the leakage of gases and vapors into the system. It is necessary to monitor the leak rate periodically to maintain the integrity of the system. It is also necessary, should the leak rate exceed specified