前言：这是一个生物药飞速发展的时代，每一个走在制药领域前沿的企业都已将目光投向生物药，生物药物因其在人体内消化代谢的特点，常常被设计为非肠道途径给药，其中最为常见的即是冻干产品。学习FDA非肠道用冻干产品现场检查的指南，有助于企业工作人员对生物药、化药等非肠道给药的冻干产品质量进行更严格控，对生产工艺进行更为合理的改进。冻干程序中有哪些影响产品质量的操作或参数？冻干工艺验证应如何展开？从哪些细节来更好的保证冻干产品的无菌？带着这些问题，我们开始学习这一指南吧~
第三期
GUIDE TO INSPECTIONS OF LYOPHILIZATION OF PARENTERALS
FDA非肠道用冻干产品检查指南
原文链接：https://www.fda.gov/ICECI/Inspections/InspectionGuides/ucm074909.htm
1. INTRODUCTION
简介
Lyophilization or freeze drying is a process in which water is removed from a product after it is frozen and placed under a vacuum, allowing the ice to change directly from solid to vapor without passing through a liquid phase. The process consists of three separate, unique, and interdependent processes; freezing, primary drying (sublimation), and secondary drying (desorption).
冻干是指药品冷冻后在真空状态下不经液态，直接从固态升华至气态，由此去除水分的作业过程。该过程包括三个彼此独立又相互依赖的步骤：冷冻、一级干燥（升华）以及二级干燥（解吸附）。
The advantages of lyophilization include:
Ease of processing a liquid, which simplifies aseptic handling
Enhanced stability of a dry powder
Removal of water without excessive heating of the product
Enhanced product stability in a dry state
Rapid and easy dissolution of reconstituted product
冻干工艺的优点在于：液体加工方便，简化了无菌作业过程；提高了干粉的稳定性；无需过热处理就能去除产品中的水分；增强了冻干产品的稳定性及复水(溶解)性。
Disadvantages of lyophilization include:
lIncreased handling and processing time
lNeed for sterile diluent upon reconstitution
lCost and complexity of equipment
冻干工艺的缺点在于：
l加工处理过程所需时间长；
l复水时需用无菌稀释液；
l设备复杂且生产成本高。
The lyophilization process generally includes the following steps:
lDissolving the drug and excipients in a suitable solvent, generally water for injection (WFI).
lSterilizing the bulk solution by passing it through a 0.22 micron bacteria-retentive filter.
lFilling into individual sterile containers and partially stoppering the containers under aseptic conditions.
lTransporting the partially stoppered containers to the lyophilizer and loading into the chamber under aseptic conditions.
lFreezing the solution by placing the partially stoppered containers on cooled shelves in a freeze-drying chamber or pre-freezing in another chamber.
lApplying a vacuum to the chamber and heating the shelves in order to evaporate the water from the frozen state.
lComplete stoppering of the vials usually by hydraulic or screw rod stoppering mechanisms installed in the lyophilizers.
冻干工艺一般包括以下步骤：
l将药品和赋形剂溶解于适当的溶剂中，通常为注射用水。
l将药液通过0.22μm的微孔除菌过滤器，以此灭菌。
l灌装到各个已灭菌的容器中，并在无菌条件下半压塞。
l在无菌条件下将半压塞的容器转移至冻干腔室内。
l冷冻该溶液：把半压塞的容器置于冻干腔室的冷冻搁板上或在另一腔室内作预冷冻。
l将腔室抽真空并将冷冻搁板升温，以便在冷冻状态下通过蒸发除去水分。
l全压塞密封：通常由安装在冻干机内的液压式或螺杆式压塞装置来完成。
There are many new parenteral products, including anti-infectives, biotechnology derived products, and in-vitro diagnostics which are manufactured as lyophilized products. Additionally, inspections have disclosed potency, sterility and stability problems associated with the manufacture and control of lyophilized products. In order to provide guidance and information to investigators, some industry procedures and deficiencies associated with lyophilized products are identified in this Inspection Guide.
现在有许多新的非肠道药品均采用冻干工艺来加工，如抗感染药物、生物制剂、体外诊断用药等等。调查及研究也已发现与冻干工艺及其控制相关的冻干剂在疗效、无菌性和稳定性方面存在问题。为此，本检查指南对冻干产品有关的生产工艺和缺陷进行了讨论，以便为调研人员提供这方面的信息和指导。
It is recognized that there is complex technology associated with the manufacture and control of a lyophilized pharmaceutical dosage form. Some of the important aspects of these operations include: the formulation of solutions; filling of vials and validation of the filling operation; sterilization and engineering aspects of the lyophilizer; scale-up and validation of the lyophilization cycle; and testing of the end product. This discussion will address some of the problems associated with the manufacture and control of a lyophilized dosage form.
人们已认识到冻干药品的生产和控制是一项很复杂的工艺技术。冻干作业的重要内容包括药液的制备；西林瓶灌装及其验证；冻干机的灭菌和设计；冻干工艺的放大生产（批量放大）及其验证；最终产品的检验等等。冻干剂成品生产和控制相关的问题是本文的重点内容。
2. PRODUCT TYPE/FORMULATION
药品种类和配方
Products are manufactured in the lyophilized form due to their instability when in solution. Many of the antibiotics, such as some of the semi-synthetic penicillins, cephalosporins, and also some of the salts of erythromycin, doxycycline and chloramphenicol are made by the lyophilization process. Because they are antibiotics, low bioburden of these formulations would be expected at the time of batching. However, some of the other dosage forms that are lyophilized, such as hydrocortisone sodium succinate, methylprednisolone sodium succinate and many of the biotechnology derived products, have no antibacterial effect when in solution.
药品需经冻干是因为其溶液状态下的不稳定性。有许多抗生素，如某些半合成的青霉素、头孢菌素和红霉素、强力霉素、氯霉素的盐类都是由冻干工艺制造的。因为是抗生素，可以预计它们在生产过程中的带菌量很低。然而，其他一类冻干剂，如氢化可的松琥珀酸钠盐、甲基泼尼松龙琥珀酸钠盐及许多生物制品在溶液状态下却毫无抗菌作用。
For these types of products, bioburden should be minimal and the bioburden should be determined prior to sterilization of these bulk solutions prior to filling. Obviously, the batching or compounding of these bulk solutions should be controlled in order to prevent any potential increase in microbiological levels that may occur up to the time that the bulk solutions are filtered (sterilized). The concern with any microbiological level is the possible increase in endotoxins that may develop. Good practice for the compounding of lyophilized products would also include batching in a controlled environment and in sealed tanks, particularly if the solution is to be held for any length of time prior to sterilization.
应尽量降低这类药品的带菌量，并在除菌过滤前（灌装前）对药液的带菌量进行测定。显然，配料或药液的配制应严加控制，以防止药液在除菌过滤前可能出现的微生物污染程度的增加。因药液内毒素的增加与微生物污染的严重程度有关。冻干剂生产与质量控制规范要求“在严格控制的环境和密封系统中生产”，对于除菌过滤前需保持一段时间的药液来说，这点尤为重要。
In some cases, manufacturers have performed bioburden testing on bulk solutions after prefiltration and prior to final filtration. While the testing of such solutions may be meaningful in determining the bioburden for sterilization, it does not provide any information regarding the potential formation or presence of endotoxins. While the testing of 0.1 ml samples by LAL methods of bulk solution for endotoxins is of value, testing of at least 100 ml size samples prior to prefiltration, particularly for the presence of gram negative organisms, would be of greater value in evaluating the process. For example, the presence of Pseudomonas sp. in the bioburden of a bulk solution has been identified as an objectionable condition.
有时候，药厂在预过滤后最终过滤前对药液进行带菌量测试。这种测定对于确定该溶液在除菌过滤时的带菌量也许有用，但是它无法提供内毒素形成以及污染水平的信息。取0.1mL样品用鲎试剂法测定其内毒素量是极为有益的，在预过滤前至少取100mL的样品进行检验(尤其是在有革兰氏阴性菌存在时)对工艺评价是至关重要的。例如：在药液中检出假单孢菌属一直被认为是将产品作报废处理的情况之一。
3. FILLING
灌封
The filling of vials that are to be lyophilized has some problems that are somewhat unique. The stopper is placed on top of the vial and is ultimately seated in the lyophilizer. As a result the contents of the vial are subject to contamination until they are actually sealed.
从某种程度上讲冻干产品的西林瓶灌封有一些颇为特殊的问题。将胶塞加到西林瓶顶部，并最终在冻干机内压塞封口，西林瓶的药液或冻干品在最终密封以前始终存在被污染的风险。
Validation of filling operations should include media fills and the sampling of critical surfaces and air during active filling (dynamic conditions).
灌封的验证应包括：培养基灌封、关键表面取样及在灌封过程中空气取样（动态条件下）。
Because of the active involvement of people in filling and aseptic manipulations, an environmental program should also include an evaluation of microbiological levels on people working in aseptic processing areas. One method of evaluation of the training of operators working in aseptic processing facilities includes the surface monitoring of gloves and/or gowns on a daily basis. Manufacturers are actively sampling the surfaces of personnel working in aseptic processing areas. A reference which provides for this type of monitoring is the USP XXII discussion of the Interpretation of Sterility Test Results. It states under the heading of "Interpretation of Quality Control Tests" that review consideration should be paid to environmental control data, including...microbial monitoring, records of operators, gowns, gloves, and garbing practices. In those situations in which manufacturers have failed to perform some type of personnel monitoring, or monitoring has shown unacceptable levels of contamination, regulatory situations have resulted.
由于人员积极地参与了灌封和无菌作业，因此环境监测计划中应包括对无菌作业区操作人员微生物污染水平的评价。其方法之一是根据日常生产中手套及工作服表面的微生物监测数据来评价无菌区操作人员接受培训的程度。现在许多药厂对无菌区的操作人员取表面样进行监控持积极态度。美国药典XXII 中“无菌检查结果解释”的讨论可以作为这类监控的一个参考材料。在“质量检测试验结果解释”项下提到，对数据进行检查回顾时，应注意环境监控结果，如微生物监测、操作人员的记录、工作服、手套及着装的实践等等。如药厂不对人员进行此类监控或出现监控结果超标的情况，必然会产生各种违规状况。
Typically, vials to be lyophilized are partially stoppered by machine. However, some filling lines have been noted which utilize an operator to place each stopper on top of the vial by hand. At this time, it would seem that it would be difficult for a manufacturer to justify a hand-stoppering operation, even if sterile forceps are employed, in any type of operation other than filling a clinical batch or very small number of units. Significant regulatory situations have resulted when some manufacturers have hand-stoppered vials. Again, the concern is the immediate avenue of contamination offered by the operator. It is well recognized that people are the major source of contamination in an aseptic processing filling operation. The longer a person works in an aseptic operation, the more microorganisms will be shed and the greater the probability of contamination.
待冻干的西林瓶通常由机器进行半压塞。然而，我们已经注意到有些灌封线是人工加塞的。除了灌封临床试验用的产品或灌封的瓶数极少这二种情况外，如今药厂很难为其在任何作业中采用人工加塞法的合理性进行辩护，即使他们使用无菌的镊子。一些采用人工加塞法的药厂已经出现了明显违规的状况。这里，问题的焦点仍是由操作人员所致的直接污染。人们都已充分认识到无菌灌封作业过程中，人员是最主要的污染源。—个操作人员在无菌作业区工作的时间越长，脱落的微生物就越多，污染的概率也就越大。
Once filled and partially stoppered, vials are transported and loaded into the lyophilizer. The transfer and handling, such as loading of the lyophilizer, should take place under primary barriers, such as the laminar flow hoods under which the vials were filled. Validation of this handling should also include the use media fills.
西林瓶灌装和半压塞一经完成，产品便被送入冻干机腔室内。转移和装入腔室等这类作业应在一级隔离的条件下进行，例如西林瓶的灌装应在层流(单向流)罩下进行。此类操作的验证也须采用培养基灌封试验。
Regarding the filling of sterile media, there are some manufacturers who carry out a partial lyophilization cycle and freeze the media. While this could seem to greater mimic the process, the freezing of media could reduce microbial levels of some contaminants. Since the purpose of the media fill is to evaluate and justify the aseptic capabilities of the process, the people and the system, the possible reduction of microbiological levels after aseptic manipulation by freezing would not be warranted. The purpose of a media fill is not to determine the lethality of freezing and its effect on any microbial contaminants that might be present.
就无菌培养基灌装而言，有些药厂将灌装的培养基作“部分冷冻干燥处理”。这样做似乎更能模拟生产的工艺过程，但冷冻会降低培养基的微生物污染水平。因为培养基灌装试验的目的是证明及评价工艺、人员和系统造就“无菌”的能力，冷冻有可能使无菌作业后的微生物污染水平下降，因而是没有道理的。培养基灌装试验的目的不是为了确定冷冻的灭菌率或其对任何可能存在的污染菌的影响。
In an effort to identify the particular sections of filling and aseptic manipulation that might introduce contamination, several manufacturers have resorted to expanded media fills. That is, they have filled approximately 9000 vials during a media fill and segmented the fill into three stages. One stage has included filling of 3000 vials and 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.
为了尽可能弄清灌装和无菌作业到底在哪些环节可能会引入污染，有些药厂采用扩大培养基灌装试验的办法。他们将灌装约9000瓶培养基的过程分为三个阶段。第一阶段灌装3000瓶，在灌装线上压塞；第二阶段灌装3000瓶，运至冻干机后压塞；第三阶段灌装3000瓶，放入冻干机，经氮气流“风淋一阵子”后压塞。因为冻干机的灭菌和用于回灌充氮(消除真空)的氮气系统的灭菌需分别进行验证，因此，培养基灌封试验应当主要是验证灌封、转运和装载等无菌作业。
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.
当工艺设计能力小于3000瓶(尤其是临床试验用药)时，经常会碰到这样一个问题，即培养基灌装试验需要灌装多少瓶？让我们再回到培养基灌装试验的目的上来回答，该试验的目的是确保药品在正常运行的条件下的无菌作业过程中不受污染，所以当最大批量小于3000瓶时，培养基灌装的数量可取其批量。
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.
灌装后的半成品通常是放在金属托盘上转移到冻干机的腔室中去的。此后，将托盘抽掉，半成品便直接置于冻干机的搁板上。检查上也碰到过这样的情况，有的药厂把半成品放入冻干机后并不抽去托盘。遗憾的是，曾经有一个厂发生了因托盘变形引起某—批号部分产品水分超标的问题。
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.
在西林瓶转入冻干机的过程中，因为尚未密封，所以还存在受污染的风险。在检查和验收新设施时，在冻干机的转运和装载区如发现没有适当的层流保护或没有—级(直接)隔离设施，可作条件不合格论。有一药厂设计了—种层流车作为补救措施，用来把西林瓶从灌装线运至冻干机。另有些建新设施的药厂把灌装线靠近冻干机安装，并且从灌装线到冻干机安装了一级屏障。
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，有一药厂在灌装线和冻干机之间安装了一个垂直的层流罩。开始时因送风的流速高而回风不足，使培养基灌装发生污染问题。经分析系气流冲击地面反弹而引起的污染。幸好，从培养基灌装试验和烟尘试验获得了足够的数据，以致这个偏差在投产前得以纠正。典型的冻干工艺往往包括西林瓶在腔室内的全压塞。
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. 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.
另一个与灌装作业有关的问题是能否保证装量的准确性。很显然，装量不足时，瓶中药效也低。和粉针或液体制剂不同，对活性成分以毫克计的生物制剂而言，灌装量偏低在冻干之后并不十分明显，但装量不足对于临床来讲，则很可能是非常严重的偏差。例如，我们在检查一家制药企业的冻干灌装作业时，发现其装量有问题，灌装机的口与灌装器错位，因而出现药液溅出和装量不足的偏差。检查时还发现，有些装量不足的西林瓶被转入了冻干机内，据此将该批产品作报废处理。
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.
有时，我们也看到这样的情况，即监测装量的生产工人只记录调整装量后的数据。所以，作为一个好的规范化的质量保证计划不仅应规定装量检查的频率，如每15分钟测一次，而且应包括出现装量过多或不足时，如何分别将它们分离开来的条款。
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.
还有一些不常用的灌装作业尚没作过讨论。例如托盘冻干，而不是灌装入西林瓶后进行冻干。就现有的技术而言，很难判断这种工艺是否适用于某个无菌产品。
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.)
双腔式西林瓶的无菌作业有更多的要求。培养基灌装试验时两个腔室均需灌装。另外，西林瓶中的稀释剂中应含有防腐剂(如无防腐剂，稀释剂的灌装就类似于培养基灌装了。在这种情况下，污染率应当为0)。
4. LYOPHILIZATION CYCLE AND CONTROLS
冻干程序及其控制
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.
将冻干机灭菌并在无菌条件下装载后，第一步作业是将药液冷冻。在某些程序中，搁板已经预冷到了冷冻药液所需的低温；但在另一些程序中，将药瓶放到搁板上后再降温至所需温度。在前一种预冷程序中，装载前搁板上是否结冰至关重要。搁板上的冰块能使瓶口的胶塞部分或全部封口，从而使产品无法冻干。近期发生过一例投诉，说药品是液状而没冻干，究其原因即是冻干程序开始前部分药瓶已经封口。遗憾的是，这家公司所作100％的目检并没有检出这些有缺陷的产品。
Typically, the product is frozen at a temperature well below the eutectic point.
一般而言，药品的冻干温度比其共熔点温度要低得多。
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.
冻干程序（包括冷冻过程）的变更或其冻干批量的放大已出现过一些问题。研究表明冷冻的速率和方式会影响冻干制品的质量。例如缓慢冷冻可导致大冰晶的形成，使空隙相对增大，有利于水蒸汽在升华时逸出；另一方面，缓慢冷冻易引起组分浓度的变化。同时冷冻的速率和方式还会影响药品的物理性质（多晶型）。
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.
冷冻后在一级干燥时将产品温度至少控制在共熔点以下4至5度较为理想。显然，药厂首先应该知道药品的共熔点，并有合适的仪器以保证产品温度分布的均匀性。冻干机也应有必要的仪表来控制并记录关键的工艺参数，如搁板温度、产品温度、冷凝器温度，腔室真空度和冷凝器的真空度。生产的规程应提供—个产品冻干程序相关的时间、温度和真空度的控制范围。监测产品温度对于偏离正常程序的作业(如停电或设备故障)尤为重要。
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.
冻干程序的机电控制采用凸轮式记录控制器，但新一代的机组采用微机控制。微机控制程序的一个基本要求是流程图或逻辑图。冻干程序的微机控制通常只是在开始时需要操作人员，如将半成品装入腔室、在半成品中插入温度探头、输入程序参数如冷冻的搁板温度、产品冷冻温度、冷冻持续时间；一级干燥时搁板温度、腔室真空度，抽真空时的产品温度；二级干燥时的搁板温度及二级干燥的时间。
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.
在一些情况下，药厂不得不在运行中不断进行调整，在此条件下，冻干程序被认为是未经验证的。
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.
冻干机软件系统的验证与其他工艺过程的验证遵循相同的准则，它包括软件的开发、修订及安全性。《药品生产微机系统检查指南》中对计算机系统可能会出现的问题作了讨论。《软件开发检验指南》是一本对软件要求进行详细回顾的参考资料。
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.
多种原因可引起冻干机的泄漏。和任何一个有真空度的腔室一样，空气进入即发生泄漏。另一类泄漏源是系统冷冻干燥所需的各种介质，包括在搁板中循环并用来加热和冷却产品的导热介质、冷阱(冷凝器)中用来冷却水蒸汽的致冷剂及可能从真空泵系统返回过来的油蒸汽。
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 limits, to determine the actual leak site for purposes of repair.
以上的任何一种因素或几种综合的因素都能造成气体或蒸汽泄露到系统。因此有必要定期监测泄漏率并维护系统的完整性。当泄漏率超过规定的限度时，必须确定实际泄漏点以便整修。
Thus, it would be beneficial to perform a leak test at some time after sterilization, possibly at the beginning of the cycle or prior to stoppering. The time and frequency for performing the leak test will vary and will depend on the data developed during the cycle validation. The pressure rise found acceptable at validation should be used to determine the acceptable pressure rise during production. A limit and what action is to be taken if excessive leakage is found should be addressed in some type of operating document.
因此，在灭菌后，在程序开始时或压塞前的某个时候进行泄漏检查是非常有益的。进行泄漏检查的时间和频率应根据程序验证中获得的数据来确定。验证中真空度减弱可接受的限度，可作为正常生产时控制的标准。应有某种操作规程来阐明泄漏允许的限度及超标后应采取的措施。
In order to minimize oil vapor migration, some lyophilizers are designed with a tortuous path between the vacuum pump and chamber. For example, one fabricator installed an oil trap in the line between the vacuum pump and chamber in a lyophilizer with an internal condenser. Leakage can also be identified by sampling surfaces in the chamber after lyophilization for contaminants. One could conclude that if contamination is found on a chamber surface after lyophilization, then dosage units in the chamber could also be contaminated. It is a good practice as part of the validation of cleaning of the lyophilization chamber to sample the surfaces both before and after cleaning.
为了减少油蒸汽进入腔室，许多冻干机在真空泵和腔室间的通道设计成曲折状。有一家生产厂在冻干机(具内接式冷凝器)的真空泵与腔室之间安装了一个油类捕集器。泄漏可在冻干程序结束后通过对腔室内表面取样加以确认。如果腔室表面已被污染，则可以得出腔室内每瓶药物亦被污染的结论。作为冻干机腔室清洁验证的一项内容，有必要在腔室清洁前后分别对其表面取样。
Because of the lengthy cycle runs and strain on machinery, it is not unusual to see equipment malfunction or fail during a lyophilization cycle. There should be provisions in place for the corrective action to be taken when these atypical situations occur. In addition to documentation of the malfunction, there should be an evaluation of the possible effects on the product (e.g., partial or complete meltback. Refer to subsequent discussion). Merely testing samples after the lyophilization cycle is concluded may be insufficient to justify the release of the remaining units. For example, the leakage of chamber shelf fluid into the chamber or a break in sterility would be cause for rejection of the batch.
由于程序的时间长以及机器是在苛刻的条件下运行，在冻干过程中发生设备故障或停止运行是不足为奇的。在生产的现场应当有在出现异常情况时如何采取纠正措施的规程。除故障相关的内容要有正式记录外，还应对产品可能受到的影响（如局部或完全熔化，下面讨论）作出评价。仅仅在冻干程序结束后对样品进行测试不足以证明除样品外的其他产品是否可准于合格，例如腔室搁板的泄漏和无菌条件的破坏均可成为产品报废的原因。
The review of Preventive Maintenance Logs, as well as Quality Assurance Alert Notices, Discrepancy Reports, and Investigation Reports will help to identify problem batches when there are equipment malfunctions or power failures. It is recommended that these records be reviewed early in the inspection.
当发生设备故障或停电时，检查预防性维修记录、质量保证黄牌警告、数额核对偏差报告和调查报告对分析确认有偏差的批是十分有益的。建议在检查时先系统审查这些记录。
5. CYCLE VALIDATION
周期性验证
Many manufacturers file (in applications) their normal lyophilization cycles and validate the lyophilization process based on these cycles. Unfortunately, such data would be of little value to substantiate shorter or abnormal cycles. In some cases, manufacturers are unaware of the eutectic point. It would be difficult for a manufacturer to evaluate partial or abnormal cycles without knowing the eutectic point and the cycle parameters needed to facilitate primary drying.
许多药厂在申报材料中上报了常规生产的冻干程序，冻干工艺的验证也是依此为基础的。但这些资料并不适用于较短的程序或非常规程序。一些情况下，药厂并不知道共熔点，如不知道共熔点和—级干燥所需的程序参数，他们对程序的某—部分或非正常的程序就难以作出评价。
Scale-up for the lyophilized product requires a knowledge of the many variables that may have an effect on the product. Some of the variables would include freezing rate and temperature ramping rate. As with the scale-up of other drug products, there should be a development report that discusses the process and logic for the cycle. Probably more so than any other product, scale-up of the lyophilization cycle is very difficult.
扩大冻干产品的批量需了解许多可能对产品造成影响的变量。这些变量包括冷冻速度和温度梯度变化的速率。同其他药品放大生产一样，应有一个开发报告来讨论程序的工艺及其逻辑性。与其他产品相比，冻干剂生产的放大更为复杂，开发报告更是必不可少的。
There are some manufacturers that market multiple strengths, vial sizes and have different batch sizes. It is conceivable and probable that each will have its own cycle parameters. A manufacturer that has one cycle for multiple strengths of the same product probably has done a poor job of developing the cycle and probably has not adequately validated their process. Investigators should review the reports and data that support the filed lyophilization cycle.
有些药厂将不同剂量、不同规格西林瓶冻干剂产品投放市场，每批的量也各不相同，可以想像这些产品应有不同的程序及各自的参数。同一品种不同剂量的产品仅使用—个冻干程序的药厂在冻干程序的开发方面所作的工作是很不够的，很可能他们还没有对他们的工艺进行过适当的验证。研究人员应该对生效的冻干程序相关的报告和数据进行系统的检查。
6. LYOPHILIZER STERILIZATION/DESIGN
冻干机灭菌和设计
The sterilization of the lyophilizer is one of the more frequently encountered problems noted during inspections. Some of the older lyophilizers cannot tolerate steam under pressure, and sterilization is marginal at best. These lyophilizers can only have their inside surfaces wiped with a chemical agent that may be a sterilant but usually has been found to be a sanitizing agent. Unfortunately, piping such as that for the administration of inert gas (usually nitrogen) and sterile air for backfill or vacuum break is often inaccessible to such surface "sterilization" or treatment. It would seem very difficult for a manufacturer to be able to demonstrate satisfactory validation of sterilization of a lyophilizer by chemical "treatment".
冻干机的灭菌是在检查过程中最为常见的一个难题。一些老式的冻干机经受不住蒸汽灭菌，灭菌最好的也只是在勉强合格的水平上。这些冻干机只能用一种化学灭菌剂对其内表面擦拭，而这类灭菌剂只不过是一种消毒剂。但引入惰性气体(如氮气)及消除真空用无菌空气的管道往往是无法用此种“表面灭菌”法加以灭菌的。看来，要让一个药厂证明化学擦试法对冻干机灭菌的验证有满意的结果是极为困难的。
Another method of sterilization that has been practiced is the use of gaseous ethylene oxide. As with any ethylene oxide treatment, humidification is necessary. Providing a method for introducing the sterile moisture with uniformity has been found to be difficult.
另—种灭菌方法是使用环氧乙烷气体。同任何环氧乙烷灭菌处理的要求—样，加湿是个必要的条件。但要提供一个均匀地引入无菌湿气的方法一直是个难题。
A manufacturer has been observed employing Water for Injection as a final wash or rinse of the lyophilizer. While the chamber was wet, it was then ethylene oxide gas sterilized. As discussed above, this may be satisfactory for the chamber but inadequate for associated plumbing.
我们检查到有一个药厂以注射用水作为最终清洗剂淋洗冻干机，当腔室是湿的时候，用环氧乙烷灭菌，正如上面已讨论的那样，此条件下腔室可得到较好的灭菌效果，但对相关的管路系统而言，这是不够的。
Another problem associated with ethylene oxide is the residue. One manufacturer had a common ethylene oxide/nitrogen supply line to a number of lyophilizers connected in parallel to the system. Thus, there could be some ethylene oxide in the nitrogen supply line during the backfilling step. Obviously, this type of system is objectionable.
与环氧乙烷灭菌相关的问题是它的残留物。有一个药厂的环氧乙烷与氮气的供气管道是一共用管路，此管路以并联的方式与几个冻干机相联。这样，在用氮气回灌做去真空处理时可能有一些环氧乙烷残留在供气管中，这样的系统应予否定。
A generally recognized acceptable method of sterilizing the lyophilizer is through the use of moist steam under pressure. Sterilization procedures should parallel that of an autoclave, and a typical system should include two independent temperature sensing systems. One would be used to control and record temperatures of the cycle as with sterilizers, and the other would be in the cold spot of the chamber. As with autoclaves, lyophilizers should have drains with atmospheric breaks to prevent back siphonage.
一个获得普遍认可的冻干机的灭菌方法是一定压力下的湿热灭菌法。灭菌程序同高压灭菌釜灭菌相似。它通常包括两个独立的测温系统，一个用于控制和记录灭菌程序的温度，另一个置于腔室的冷点。同灭菌釜—样，冻干机排水口与地漏间应有空气断口，能防止虹吸。
As discussed, there should also be provisions for sterilizing the inert gas or air and the supply lines. Some manufacturers have chosen to locate the sterilizing filters in a port of the chamber. The port is steam sterilized when the chamber is sterilized, and then the sterilizing filter, previously sterilized, is aseptically connected to the chamber. Some manufacturers have chosen to sterilize the filter and downstream piping to the chamber in place. Typical sterilization-in-place of filters may require steaming of both to obtain sufficient temperatures. In this type of system, there should be provisions for removing and/or draining condensate. The failure to sterilize nitrogen and air filters and the piping downstream leading into the chamber has been identified as a problem on a number of inspections.
正如所讨论的那样，应制订惰性气体、空气及其管道系统灭菌的规程。一些厂家采用了在腔室的进气口安装除菌过滤器的办法，将腔室进行湿热灭菌时，进气口也灭了菌，然后将事先灭好菌的除菌过滤器以无菌方式安装在进气口，使管路系统与腔室相联。另一些厂家则采用对过滤器及进气口下游管道进行在线灭菌的手段。通常，在线灭菌时，二者均须达到足够高的温度。在这类系统中冷凝水的排放或清除应有书面的规定。在检查中发现，氮气、空气过滤器及进气口管道灭菌不完全的现象是一个带普遍性的问题。
Since these filters are used to sterilize inert gas and/or air, there should be some assurance of their integrity. Some inspections have disclosed a lack of integrity testing of the inert gas and/or air filter. The question is frequently asked how often should the vent filter be tested for integrity? As with many decisions made by manufacturers, there is a level of risk associated with the operation, process or system, which only the manufacturer can decide. If the sterilizing filter is found to pass the integrity test after several uses or batches, then one could claim its integrity for the previous batches. However, if it is only tested after several batches have been processed and if found to fail the integrity test, then one could question the sterility of all of the previous batches processed. In an effort to minimize this risk, some manufacturers have resorted to redundant filtration.
由于这些过滤器是惰性气体和空气灭菌过滤用的，因此，应确保它们的完好性。检查中发现，惰性气体和空气过滤器完好性试验往往没做。常常有人问我们这样的问题，即空气过滤器完好性试验应多久做一次？同药厂做出各种各样的决定一样，风险的大小与操作、工艺及系统有关，只有药厂才能对此做出决定。如果灭菌过滤器在数次使用或生产数批产品后完好性试验结果仍合格，则可以说明以前生产的批所用的过滤器是完好的。但如果在使用几次后发现过滤器的完好性试验不合格，那么在此前生产的全部产品的无菌性就成为问题，为了减少这种风险，一些厂家采用了使用串联过滤的办法。
For most cycles, stoppering occurs within the lyophilizer. Typically, the lyophilizer has some type of rod or rods (ram) which enter the immediate chamber at the time of stoppering. Once the rod enters the chamber, there is the potential for contamination of the chamber. However, since the vials are stoppered, there is no avenue for contamination of the vials in the chamber which are now stoppered. Generally, lyophilizers should be sterilized after each cycle because of the potential for contamination of the shelf support rods. Additionally, the physical act of removing vials and cleaning the chamber can increase levels of contamination.
大多数程序的压塞作业是在冻干机内完成的。冻干机通常都有某种类型的螺旋杆，在加塞时直接进入腔室。—旦螺旋杆进入腔室，就有可能会给腔室带来污染。然而，由于西林瓶已全压塞，污染西林瓶的途径亦不复存在。由于搁板的支架存在污染的可能，每次冻干程序结束后冻干机一般应灭菌。此外，从腔室取出产品并进行清洗的过程也可导致污染的增加。
In some of the larger units, the shelves are collapsed after sterilization to facilitate loading. Obviously, the portions of the ram entering the chamber to collapse the shelves enters from a non-sterile area. Attempts to minimize contamination have included wiping the ram with a sanitizing agent prior to loading. Control aspects have included testing the ram for microbiological contamination, testing it for residues of hydraulic fluid, and testing the fluid for its bacteriostatic effectiveness. One lyophilizer fabricator has proposed developing a flexible "skirt" to cover the ram.
对体积较大的产品而言，灭菌后，可将搁板折叠起来以方便装瓶。显然，用来折叠搁板的推杆有部分是从非灭菌区伸入腔室的，为了减少污染，在装瓶前应用消毒剂擦拭推杆。此外，测定推杆的微生物污染、测定液压油的残渣及测定该液体的抑菌有效性均应作为监控的内容。有一冻干机的制造商提议要研制一种可伸缩的“罩”以便将推杆隔离。
In addition to microbiological concerns with hydraulic fluid, there is also the concern with product contamination.
除了液压油的微生物污染外，还应考虑它对产品的污染问题。
During steam sterilization of the chamber, there should be space between shelves that permit passage of free flowing steam. Some manufacturers have placed "spacers" between shelves to prevent their total collapse. Others have resorted to a two phase sterilization of the chamber. The initial phase provides for sterilization of the shelves when they are separated. The second phase provides for sterilization of the chamber and piston with the shelves collapsed.
在用蒸汽灭菌腔室过程中，搁板间应有空间使蒸汽自由通过。—些制药厂家在搁板间放置“垫片”以防止它们全部折叠起来。另—些药厂对腔室的灭菌采用二次灭菌法：先对搁板（分开状态）作—次灭菌，然后在活塞推杆将搁板全部折叠好后再对腔室灭菌一次。
Typically, biological indicators are used in lyophilizers to validate the steam sterilization cycle. One manufacturer of a Biopharmaceutical product was found to have a positive biological indicator after sterilization at 121°C for 45 minutes. During the chamber sterilization, trays used to transport vials from the filling line to the chamber were also sterilized. The trays were sterilized in an inverted position on shelves in the chamber. It is believed that the positive biological indicator is the result of poor steam penetration under these trays.
通常采用生物指示剂来验证冻干机的蒸汽灭菌程序。有一个生物制药厂在121°C灭菌45分钟后仍发现生物指示剂出现阳性结果。在腔室灭菌过程中，用以将半成品从灌装线送往腔室的托盘也灭了菌，但托盘是倒置在腔室的搁板上灭菌的，生物指示剂呈现阳性结果很可能就是由此引起热穿透变差所致。
The sterilization of condensers is also a major issue that warrants discussion. Most of the newer units provide for the capability of sterilization of the condenser along with the chamber, even if the condenser is external to the chamber. This provides a greater assurance of sterility, particularly in those situations in which there is some equipment malfunction and the vacuum in the chamber is deeper than in the condenser.
冷凝器的灭菌也是一个值得讨论的要点。即使冷凝器是安装在腔室外的，大多数新型冻干机可以将冷凝器与腔室同时灭菌。这样获得的无菌保证更可靠，在设备出现故障、腔室的真空度比冷凝器真空度大的情况下更是如此。
Malfunctions that can occur, which would indicate that sterilization of the condenser is warranted, include vacuum pump breakdown, refrigeration system failures and the potential for contamination by the large valve between the condenser and chamber. This is particularly true for those units that have separate vacuum pumps for both the condenser and chamber. When there are problems with the systems in the lyophilizer, contamination could migrate from the condenser back to the chamber. It is recognized that the condenser is not able to be sterilized in many of the older units, and this represents a major problem, particularly in those cycles in which there is some equipment and/or operator failure.
出现故障时，如真空泵的损坏、冷却系统的失灵、冷凝器和腔室间大阀门可能产生污染等，冷凝器的灭菌就显得十分必要。这一点对腔室和冷凝器有单独真空泵的系统尤为重要，因为当冻干机系统出现故障时，污染可以从冷凝器转移到腔室。许多老式冻干机的冷凝器不能灭菌，当设备或操作出现问题时，它将成为一个严重的问题。
As referenced above, leakage during a lyophilization cycle can occur, and the door seal or gasket presents an avenue of entry for contaminants. For example, in an inspection, it was noted that during steam sterilization of a lyophilizer, steam was leaking from the unit. If steam could leak from a unit during sterilization, air could possibly enter the chamber during lyophilization.
如上所述，在一个冻干程序运行过程中有可能发生泄漏，门的密封圈或垫圈可成为污染物进入的途径。在一次检查中，在冻干机蒸汽灭菌过程中发现蒸汽泄漏。既然灭菌时蒸汽能从冻干机泄漏出来，则在冻干过程中，空气必然也能进入其中。
Some of the newer lyophilizers have double doors - one for loading and the other for unloading. The typical single door lyophilizer opens in the clean area only, and contamination between loads would be minimal. This clean area, previously discussed, represents a critical processing area for a product made by aseptic processing. In most units, only the piston raising/lowering shelves is the source of contamination. For a double door system unloading the lyophilizer in a non-sterile environment, other problems may occur. The non-sterile environment presents a direct avenue of contamination of the chamber when unloading, and door controls similar to double door sterilizers should be in place.
一些新式的冻干机有两个门，分别用于产品的装和卸。单门冻干机的门一般只在洁净区打开，因此对产品污染的风险很小。如先前所述。这个洁净区是无菌生产的关键作业区。对大多数冻干机而言，升降搁板用的活塞是唯一的污染源。对于双门冻干机而言，成品是在非洁净区卸瓶的，因此会出现其他方面的问题。非洁净区环境在卸瓶时成了污染腔室的直接途径，因此，其门的控制应与双门灭菌釜相似。
Obviously, the lyophilizer chamber is to be sterilized between batches because of the direct means of contamination. A problem which may be significant is that of leakage through the door seal. For the single door unit, leakage prior to stoppering around the door seal is not a major problem from a sterility concern, because single door units only open into sterile areas. However, leakage from a door gasket or seal from a non-sterile area would present a significant microbiological problem. In order to minimize the potential for contamination, it is recommended that the lyophilizers be unloaded in a clean room area to minimize contamination. For example, in an inspection of a new manufacturing facility, it was noted that the unloading area for double door units was a clean room, with the condenser located below the chamber on a lower level.
显然，冻干机腔室在各批之间应进行灭菌以避免直接污染。门密封处的泄漏有可能成为—个十分严重的问题。对于单门冻干机而言，从无菌保证的观点去看，压塞前密封门的泄漏并不是大问题，因为门只在无菌区打开。然而，非洁净区门密封圈或垫圈的泄漏将引起严重的微生物污染问题。为了尽量减少污染，冻干机内成品的卸瓶程序应在洁净区进行。在检查一个新厂时，我们曾注意到双门冻干机的卸货口也设在洁净区，冷凝器则安装在腔室的下方。
After steam sterilization, there is often some condensate remaining on the floor of the chamber. Some manufacturers remove this condensate through the drain line while the chamber is still pressurized after sterilization. Unfortunately, some manufacturers have allowed the chamber to come to and remain at atmospheric pressure with the drain line open. Thus, non-sterile air could contaminate the chamber through the drain line. Some manufacturers have attempted to dry the chamber by blowing sterile nitrogen gas through the chamber at a pressure above atmospheric pressure.
蒸汽灭菌后，在冻干机腔室的底部会有一些冷凝水。有些药厂在灭菌后用“加压排水法”通过地漏排放冷凝水；但有些药厂却在常压下从地漏排放。这样，非无菌空气能通过排放管污染腔室。还有些厂试图用高于大气压的无菌氮气吹入腔室，使其干燥。
In an inspection of a biopharmaceutical drug product, a Pseudomonas problem probably attributed to condensate after sterilization was noted. On a routine surface sample taken from a chamber shelf after sterilization and processing, a high count of Pseudomonas sp. was obtained. After sterilization and cooling when the chamber door was opened, condensate routinely spilled onto the floor from the door. A surface sample taken from the floor below the door also revealed Pseudomonas sp. contamination. Since the company believed the condensate remained in the chamber after sterilization, they repiped the chamber drain and added a line to a water seal vacuum pump.
我们在一次生物药厂的检查中，发现灭菌后的冷凝水曾引起假单孢菌属的污染，在灭菌及作业完成后在腔室搁板表面的常规取样中检查出了大量的假单孢菌属。灭菌及冷却后，将腔室门打开时，经常有冷凝液溅到地板上，在门下地板表面取样中也发现了假单孢菌属的污染。由于该厂确认了冷凝液引起的污染问题，他们改装厂腔室的排水管道，并增加了一条通向水封真空泵的管道。
7. FINISHED PRODUCT TESTING FOR LYOPHILIZED PRODUCTS
冻干产品的成品检验
There are several aspects of finished product testing which are of concern to the lyophilized dosage form. These include dose uniformity testing, moisture and stability testing, and sterility testing.
冻干剂的成品测试的内容有：装量差异、水分、稳定性考察及无菌检查。
(a) Dose Uniformity装量差异
The USP includes two types of dose uniformity testing: content uniformity and weight variation. It states that weight variation may be applied to solids, with or without added substances, that have been prepared from true solutions and freeze-dried in final containers. However, when other excipients or other additives are present, weight variation may be applied, provided there is correlation with the sample weight and potency results. For example, in the determination of potency, it is sometimes common to reconstitute and assay the entire contents of a vial without knowing the weight of the sample. Performing the assay in this manner will provide information on the label claim of a product, but without knowing the sample weight will provide no information about dose uniformity. One should correlate the potency result obtained from the assay with the weight of the sample tested.
美国药典包括两类试验：含量均匀度和重量差异。重量差异适用于由真正的溶液通过冷冻干燥制备在最终容器中的各种固体，不管在溶液中是否添加了什么组分。当其他赋形剂或添加剂存在时，只有样品重量与效价(主药量)之间存在一定相关性的条件下，重量差异的检测才是有意义的。在含量测定时，通常是在不知道样品重量的情况下将药物溶解，测得的是西林瓶中总的量。进行这类定量测定可以为产品标签上的标示量提供数据，但是，由于样品的重量、来知，故无法得到剂量均匀度的信息，应确定测得的主药含量同检品重量的相关性。
(b) Stability Testing稳定性考察
An obvious concern with the lyophilized product is the amount of moisture present in vials. The manufacturer's data for the establishment of moisture specifications for both product release and stability should be reviewed. As with other dosage forms, the expiration date and moisture limit should be established based on worst case data. That is, a manufacturer should have data that demonstrates adequate stability at the moisture specification.
西林瓶中水分显然是冻干剂的关键项目。应回顾检查药厂建立“产品放行和稳定性要求”中的水分控制限度的数据和资料。与其他制剂一样，有效期和水分控制标准应以最差的数据为基础，即药厂应确定数据能证明在该水分条件下产品有足够的稳定性。
As with immediate release potency testing, stability testing should be performed on vials with a known weight of sample. For example, testing a vial (sample) which had a higher fill weight (volume) than the average fill volume of the batch would provide a higher potency results and not represent the potency of the batch. Also, the expiration date and stability should be based on those batches with the higher moisture content. Such data should also be considered in the establishment of a moisture specification.
在作主药测定以便“准予放行”的同时，应对样品重量已知的产品作稳定性试验。要知道，对一个高于平均装量(重量或体积)的西林瓶(样品)进行测定时，测得主药含量的结果较高，但它并不代表整批产品主药的含量。此外，有效期和稳定性也应以水分含量高的批次为基础，在建立水分控制标准(限度)时，这些数据均应考虑。
For products showing a loss of potency due to aging, there are generally two potency specifications. There is a higher limit for the dosage form at the time of release. This limit is generally higher than the official USP or filed specification which is official throughout the entire expiration date period of the dosage form. The USP points out that compendial standards apply at any time in the life of the article.
对于那些主药含量随时间而下降的产品，通常主药的标准有二个。在出厂检验时的控制标准(限度)要高，比该产品有效期内美国药典或正式注册的标准要高一些，以确保美国药典所规定的“药品在其有效期的任何时间都应当符合所规定的各项标准”的要求。
Stability testing should also include provisions for the assay of aged samples and subsequent reconstitution of these aged samples for the maximum amount of time specified in the labeling. On some occasions, manufacturers have established expiration dates without performing label claim reconstitution potency assays at the various test intervals and particularly the expiration date test interval. Additionally, this stability testing of reconstituted solutions should include the most concentrated and the least concentrated reconstituted solutions. The most concentrated reconstituted solution will usually exhibit degradation at a faster rate than less concentrated solutions.
稳定性考察应对老样品定量测试并按标签上规定的最长储存时间安排的测试计划做出规定。有些药厂在没有作不同时间间隔特别是标签所规定有效期试验的条件下制订了产品的有效期。此外，稳定考察还应包括对再溶解后药液的最高和最低两个浓度的试验，因高浓度药液的降解速度一般比低浓度的高。
(c) Sterility Testing无菌检查
With respect to sterility testing of lyophilized products, there is concern with the solution used to reconstitute the lyophilized product. Although products may be labeled for reconstitution with Bacteriostatic Water For Injection, Sterile Water For Injection (WFI) should be used to reconstitute products. Because of the potential toxicities associated with Bacteriostatic Water For Injection, many hospitals only utilize WFI. Bacteriostatic Water For Injection may kill some of the vegetative cells if present as contaminants, and thus mask the true level of contamination in the dosage form.
就冻干制剂的无菌检查而言，溶解冻干剂产品的溶剂是一个必须注意的方面。尽管标签上规定应用抑菌的注射用水、无菌注射用水溶解产品。因为抑菌的注射用水中隐含着毒性，所以许多医院往往只用注射用水来溶解产品。抑菌注射用水可以杀死污染产品的某些生长态菌，从而掩盖了产品的实际污染程度。
As with other sterile products, sterility test results which show contamination on the initial test should be identified and reviewed.
和其他无菌产品的无菌检查一样，应当弄清哪些是第—次无菌检查呈现阳性结果的批号，并对结果作系统的回顾和检查。
8. FINISHED PRODUCT INSPECTION - MELTBACK
成品检查——回熔
The USP points out that it is good pharmaceutical practice to perform 100% inspection of parenteral products. This includes sterile lyophilized powders. Critical aspects would include the presence of correct volume of cake and the cake appearance. With regard to cake appearance, one of the major concerns is meltback.
美国药典指出对非经肠道药进行100%目检符合，生产质量管理规范的要求，它包括对无菌冻干粉末的目检。干饼体积及其外观是目检的重要方面，而外观的重点又在“回熔”上。 回熔是干饼损坏的表现，究其原因是发生了由固态向液态的变化，即冻干过程中的升华(从固态到气态)不完全。与之相关的问题是药物物理形成的变化和形成湿块的风险，它将导致产品不稳定性的增加和降解的加快。
Another problem may be poor solubility. Increased time for reconstitution at the user stage may result in partial loss of potency if the drug is not completely dissolved, since it is common to use in-line filters during administration to the patient.
另一个可能出现的问题是溶解度变差。在给病人输注时，由于输液器上通常装有—个过滤器，如果药物溶解不完全，溶解时间的延长可导致药效的部分丧失。
Manufacturers should be aware of the stability of lyophilized products which exhibit partial or complete meltback. Literature shows that for some products, such as the cephalosporins, that the crystalline form is more stable than the amorphous form of lyophilized product. The amorphous form may exist in the "meltback" portion of the cake where there is incomplete sublimation.
药厂应该了解这类部分回熔或完全回熔冻干产品的稳定性情况。文献表明，—些冻干产品，如头孢菌素类抗生素，其晶状体的稳定性优于无定形体。在干饼的回熔体部分可能存在无定形体，那里的升华是不完全的。
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