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6.0 Analytical Methods分析方法
It is essential to a cleaning validation program that the appropriate analytical methods are utilized.
一个清洁验证程序使用适当的分析方法是非常必要的。
Analytical methods must be appropriate in that they can adequately detect the residue(s) of concern.分析方法必须适当，能充分检测到相关残留物。
It is also important to understand what can be concluded from the analytical result (e.g., was the productnot removed or was the cleaning agent not removed?).
对能从分析结果中推断出什么的理解也是非常重要的（比如：产品没有被去除或清洗剂没有被去除？） 。
The results of testing will determine if the cleaning validation cycle is acceptable or if it needs to beredeveloped.
检测结果将决定清洁验证周期是否接受或者是否需要重新开发。
Thus, it is important to have confidence in the results.
因此，对结果的信任是非常重要的。
This section discusses how to select the appropriate assay methods, detailed information on theapplicability and use of nonspecific assays and microbial test methods,and assay method validation.
本部分讨论怎样选择合适的分析方法及其适用性的详细信息，非特定分析和微生物测试方法的使用，和分析方法验证。本部分套乱怎样选择合适的含量分析方法，实用性的详细信息，非特定含量方法和微生物测试方法以及含量方法验证。
6.1 Specific Analytical Methods
6.1 特定分析方法
Specific analytical methods are those which measure a certain residue in the presence of expectedinterferences.
特定分析方法是指用于测量存在预期干扰时某个残留的方法。
In a cleaning process for biotechnology products where the specific analyte is the active protein, suchinterferences may include degradation products and related substances, excipients, cleaning agents andcleaning agent by-products.
在生物技术产品的清洗过程中，特定分析物是活性蛋白，这样干扰可能包括降解产品和相关物质，辅料，清洗剂和产品清洗剂。
Examples of specific methods include HPLC, ELISA,SDS PAGE, and PCR.
特定方法的例子包括 HPLC,ELISA,SDSPAGE,和 PCREach
of these methods requires the use of an appropriate reference standard.
这些方法每个都需要使用一个适当参考标准。
In contrast, nonspecific analytical methods measure a general property, such as conductivity orTOC,which could be due to a variety of analytes or sources.
相对而言， 非特定分析方法测量通用属性， 比如：电导率或者 TOC， 因为分析物或者来源的多样性。
Selection of an analytical method will depend on the nature of the residue as it exists after the cleaningprocess.
一个分析方法的选择将取决于清洁过后残留物的性质。
Only if a protein (or other organic active) is not degraded during the cleaning process (surviving hightemperatures and pH extremes in an aqueous environment, for example) does it make sense to use a specific analytical method for that active.
只有当一种蛋白质（或者其他有机活性物质）在清洁过程中不能被降解（例如：在高温和极端 PH 的水环境下残存） ，使用那种物质的特定分析方法才有意义。
The advantage of using a specific analytical method in this situation is that it gives a precise measure ofthe major residue of concern – the active itself.
在这种情况下使用特定分析方法的优势是它可以给出主要相关残留--活性物本身的精确测量。
If a specific analytical method for an active protein were utilized following a cleaning process which hasbeen demonstrated to denature (degrade) that active protein, it is likely that residues of the activeprotein would be non-detectable (i.e., not measurable) by that specific analytical method.
如果一个活性蛋白质的特定分析方法在紧接着的清洁过程中被使用，这个清洁过程用来证明降解（变性）这个活性蛋白质。这个活性蛋白残留通过这个特定分析方法将很可能检测不到（不能被检测） 。
Residues of that protein would be various degraded fragments.
那个蛋白质的残留可能讲解成多种片段。
If the native protein were actually detected using a specific method for that protein, it is likely that therehad been a serious problem with the cleaning process, such as a clogged spray device causing a lack ofcoverage of that portion of the equipment surface.
如果天然的蛋白质真的用特定方法被检测到，那么清洁过程很有可能有一系列的问题，比如堵塞喷淋设备引起的设备表面喷淋覆盖不全面。
In such a case, failure would also most likely be detected by a nonspecific method and/or by visualexamination.
在这种情况下，失败也可以通过非特定方法或目视检察被检测到。
Consequently, if a specific assay method is used, a nonspecific assay method is also required, unlessstudies prove that the product is not degraded by the cleaning process.
因此，如果一个特定的含量分析方法被使用，一个非特定的分析方法也是需要的，除非研究证明清洗过程中产品没有被分解。
In biotechnology cleaning validation, specific analytical techniques such as HPLC are more likely to beused for detergents, because the surfactants or other functional materials in the detergents are not likelyto degrade in the cleaning process.
在生物技术清洁验证中，特定分析技术比如 HPLC 更倾向用于清洗剂的检测，因为表面活性剂或者清洁剂中的功能材料在清洁过程中不可能分解。
However, it should be noted that nonspecific methods can also be used for detergents and other cleaningagents.
无论怎样，必须注意非特定方法也可以用于洗涤剂或者其他清洁剂
6.2 Impact of Inactivation/Degradation of the Active
6.2 活性物质的降解/ 灭活的影响
Product inactivation means that the active protein is modified in some way such that it is no longer activeand may no longer be measurable by specific analytical methods for that native protein.
产品灭活意味着活性蛋白被某些方式改变，这样这些蛋白不再有活性和不再被这种天然蛋白质的特定分析方法测量到。
This modification usually involves degradation of the active protein into smaller fragments, but may alsoinvolve a process in which larger molecules are formed.
这个改变通常包括活性蛋白降解成小片段，但也可以包括蛋白大分子的形成过程。
A key issue for process equipment cleaning in biotechnology manufacturing is the degradation ordeactivation of the active protein during the cleaning process.
在生物技术产品生产中，工艺设备清洁的一个关键问题是活性蛋白在清洁过程中的降解或者钝化。
This is a result of cleaning processes in biotechnology utilizing hot, aqueous,alkaline and acidic cleaningsolutions.
这是生物技术产品的清洗过程使用热，水，碱和酸的清洗溶剂的结果。
Under such conditions, it is well recognized that protein actives will degrade.
在这种条件下，将要降解的蛋白质活性物是容易被识别的。
This degradation affects several issues in the cleaning and cleaning validation process.
这个降解物在清洁和清洁验证过程中的影响是多方面的。
Because of the degradation, the residues of the active protein (which are actually now residues of thedegraded active protein) are more readily rinsed away during the rinsing step of the cleaning process.
因为降解，活性蛋白的残留（实际上是活性蛋白降解物的残留）很容易在清洁过程中的淋洗这步时被淋洗掉。
This is because the degraded fragments typically have a lower molecular weight and are potentially morepolar, both conditions leading to greater water solubility.
这是因为降解的片段通常分子量小和极性更大，两种条件都导致更好的水溶解性。
A second consequence of the degradation is that it no longer is scientifically justified to have an analyticalmethod which is specific for the native protein.
降解的第二个结果是使用一个特定的天然蛋白质分析方法不再是科学合理的。
For this reason, a nonspecific method such as TOC or Total Protein is typically used to measure residues ofthe degraded active (as well as other organic molecules) in acleaning validation protocol.基于这个原因，在清洁验证方案中，一个非特定方法（如：TOC 和总蛋白）通常被用于测量降解活性物质（其他有机分子也一样）的残留。
A third consequence is that limits in bulk biotechnology manufacturing are typically not appropriatelyestablished based on a “fraction of a dose” calculation of the native protein, since the residues aredegraded fragments.
还有一个结果是生物原料生产中的限度通常不能基于天然蛋白“剂量分数”的计算建立，因为残留是降解片段。
Since residues being sampled are residues of the degraded protein, it may also make more scientific senseto perform sampling recovery studies based on recovery of the degraded fragments.
因为残留取样是降解蛋白的残留，所以执行基于降解片段回收率的取样回收研究是更加科学有意义的。
However, assuming an increase in solubility for degraded proteins, sampling recovery studies on thenative protein will typically be a worst case as compared to recovery of degraded fragments.
假如降解蛋白的溶解性增加，天然蛋白的取样回收研究与降解片段的回收相比，其通常作为最差情况。
While it is assumed in almost all cases that the active proteins or other large organic molecules producedin biotechnology manufacturing are readily degraded in hot, aqueous alkaline conditions, it is desirable todemonstrate this with a laboratory study.
几乎所有情况下，活性蛋白或者大型有机分子在生物生产中，在热，水，碱条件下非常容易降解，这只是个假设，因此需要在实验室中证明。
In such a simple “beaker” study, the bulk active protein is exposed to the conditions of the cleaningprocess, including cleaning agent concentration,temperature and time.
在这样一个简单的“烧杯”研究，活性蛋白原料暴露在清洗过程的条件下，包括清洗剂浓度，温度和时间。
At the end of that exposure time, the pH is neutralized, and the temperature is reduced.
在暴露时间的最后，PH 是中性的，温度是降低的。
The resultant solution is then analyzed for the active protein by the specific analytical procedure (such as ELISA, HPLC or a bioactivity assay).
最终溶液被特定分析程序分析其活性蛋白质（例如：ELISA,HPLC 或者生物活性含量）
In such a procedure, the ratio of protein to cleaning solution should represent the same ratio present during cleaning, or a worst-case ratio (a worst-case is a higher ratio of protein to cleaning solution).
在这个过程中，清洁溶液的蛋白质比例应等于清洁过程中蛋白质的比例，或者最差情况蛋白质的比例（最差情况是一个更高蛋白比例的清洁溶液） 。
The assay methodology for such studies must be appropriate and valid.
这个研究的含量方法学必须是合适的和经验证的。
Care needs to be exercised in performing such a study to ensure that the chemicals in the cleaningsolution do not interfere with the analytical procedure.
为了确保清洁溶液中化学成分不被分析程序干扰，在进行这个研究过程中应特别注意。
This can be addressed by having adequate controls, such as adding the active to a solution of theneutralized cleaning solution at ambient temperature.
这可以用充分控制来解决，如：向日常温度的中性清洗溶液加入活性物质。
If chemicals in the cleaning solution interfere with the specific analytical procedure, another option is toremove them by diafiltration.
如果清洗溶液的化学成分被干扰特定分析程序，另一个选择是通过透析过滤去除它们。
If it is just the surfactants in the cleaning agent that interfere, another option is to perform thedegradation study with just the equivalent amount of alkali present in the cleaning solution.
如果它只是清洗剂中的表面活性剂的干扰，另一个选择是用与清洗溶液中等量的碱进行降解研究。
Note that in many cases, cleaning in a biotechnology facility utilizes alkaline cleaning agents followed byan acidic cleaning solution.
注意在许多情况下，生物技术设施的清洁碱清洁剂在使用酸清洁溶液之后使用。
Current evidence suggests that it is the alkaline portion that is most effective in degrading active proteins.
现有的证据证明含碱的部分对降解活性蛋白非常有效。
Companies may choose to perform a degradation study only with the alkaline agent and not pursue degradation studies with the acidic solution unless the alkaline cleaning agent alone is inadequate for degradation.
公司可以选择只使用含碱试剂进行降解研究，不追求使用酸溶液进行降解研究除非单独使用含碱清洁剂对降解不充分。
6.3 Nonspecific Analytical Methods
6.3 非特定分析方法
6.3.1 Total Organic Carbon (TOC)
6.3.1 总有机碳
Most of the compounds used in biotechnology processes are of organic nature.
在生物技术过程中使用的大部分成分是有机物。
TOC can detect organic carbon with a good sensitivity in the sub-ppm range;
TOC 在亚于 PPM 范围内可以非常灵敏的检测有机碳。
however this sensitivity may still not be adequate for highly active substances.
尽管这样的灵敏性可能对于高活性物质来说仍然是不足够的。
The method can be semi-automated with an autosampler and has a short analyzing time.
这种方法可以是半自动的，用一个自动采样器和用很短的分析时间。
In contrast to specific analytical methods, TOC analyzers can detect all organic residues, including complex mixtures of compounds like cell culture media or product degraded by the cleaning process.相对于特定分析方法而言，TOC 分析仪可以检测所有有机残留，包括复杂的混合成分像细胞培养基或者清洗过程中产品讲解物。
With TOC, it is not possible to distinguish between a biotechnology product and other organic compounds present in the same sample.
使用 TOC，在相同的样品中是无法区别生物制品和其他有机物成分的。
As a consequence, all organic carbon is assumed to be product, representing a worst-case approach.因此，所有有机碳被当成产物，这是最差情况的方法
Another aspect is the potential for sample contamination with organic substances during sampling and testing, requiring well- trained personnel and clear sampling instructions.
另一方面，在取样和测试过程中存在有机物取样污染的可能性，需要经过充分培训的人员和明确的取样说明。
Special care should be taken to ensure that the sampling container does not introduceunacceptably high amounts of carbon to the sample.
必须有特别措施确保取样容器不能给样品引入不可接受数量的碳
Different TOC analyzers are commercially available. All instruments oxidize organic carbon and measure the resulting carbon dioxide.
不同的 TOC 分析仪是商业上可行的。所有仪器氧化有机碳和测量产生的二氧化碳。
When selecting a TOC instrument, care should be taken to select an instrument and instrumentparameters that are able to completely oxidize the organic carbon present.
当选择一个 TOC 仪器时，必须注意：选择一个仪器和仪器参数，其必须能够完全氧化有机碳。
The TOC method can be used for rinse and swab measurements. If used for final rinse water testing, samples can be analyzed directly.
TOC 方法可以被用于淋洗和擦拭测量。如果被用于最终淋洗水检测，样品必须能够直接被分析。
If used for swab testing, the organic carbon has to be extracted from the swab after sampling.
如果被用于擦拭测试，取样后有机碳必须能够从擦拭棒上被萃取出
It is important to consider some additional topics during TOC swab method development, such as swab material and technique.
在擦拭方法开发过程中考虑一些额外的问题是非常重要的，比如:擦拭材料和技术
Swabs should not significantly contribute carbon to the sample and should not adsorb significant amountsof the residue such that it is not released for analysis.
擦拭对于样品不是重要的碳来源， 不能吸附大量的残留成分， 分析过程中残留成分也不能释放出来。
The sampling technique (e.g., swab size and shape, swabbing pattern, swab container and extraction method) is much more complex in comparison to rinse water testing.
取样技术（如：擦拭面积和形状，擦拭方式，擦拭容器，提取方法）相对于淋洗水测试来说是及其复杂的。
The swabbing technique can have a high influence on residue recovery.
擦拭技术能对残留回收率产生极大的影响
6.3.2 Total Protein
6.3.2 总蛋白质
Several total protein assays of different sensitivity are commercially available. Assays often used are Bradford, Lowry or BCA.
一些不同灵敏度的总蛋白质含量测试方法在商业上是可行的。含量测量方法通常使用 bradford 法,lowry 法或者 BCA 法。
Total protein assays are not product specific, but specific towards a class of molecules.
总蛋白质含量不是产品特定，但是对于分子类型来说是特定的。
Total protein assays can be used if the majority of the residues are proteins.
总蛋白质含量可以被用于检测主要残留是否是蛋白质。
If proteins are just one of many residues present (e.g., cell culture fermentation), the use of an assay with a broader spectrum (e.g., TOC) should be considered.
如果蛋白质不只是许多残留（细胞培养发酵）中的一种，使用宽谱（如 TOC）测量含量可以被考虑。
One advantage of total protein assays is potential commercial availability.
测量总蛋白质含量的一个优势是具有商业上的可行性。
Different companies offer test kits and support during test implementation.
不同的公司提供测试工具和测试应用支持。
Lead times for implementing commercial test kits are typically shorter compared to in-house developed methods.
与内部开发方法相比，使用商业测试工具通常测试时间更短。
In-house methods may be developed to provide enhanced sensitivity.
内部方法可以被开发用来加强灵敏性。
Proteins often degrade (e.g., by hydrolysis during a cleaning cycle if high pH and temperatures are used).
蛋白质经常降解（比如，如果用高 PH 和温度进行清洁，清洁过程中会水解）
During assay implementation, it should be investigated if the assay still can detect protein after exposure to cleaning agents.
在含量测定方法应用过程中，暴露在清洁剂过后如果蛋白质含量仍然能被检测到，必须进行调查。
6.3.3 Conductivity
6.3.3 电导率
Conductivity measurement is a very sensitive method to detect dissociated ionic substances in water samples.
电导率测量是一种检测水样中游离离子物质非常灵敏的方法。
WFI has a conductivity of ≤ 2.4 μ S/cm at 65°C. For cleaning validation purposes,conductivity readings are expressed in milli-Siemens/cm (mS/cm) for higher concentrations (such as cleaning solutions) and micro-Siemens/cm (μS/cm) for lower concentrations (such as final rinse waters ).注射用水在 65°C 时电导率≤ 2.4 μ S/cm。
清洁验证的目标，电导率的读数单位浓度高时（清洁溶液）用 mS/cm 浓度低时（如：最终淋洗水）用(μ S/cm)。
It is often used to measure cleaning agent residues (e.g., caustic agents) and to control automated cleaning processes (e.g., CIP).
电导率经常被用来测量清洁剂残留（如：腐蚀剂）和控制自动清洗程序（如：CIP）Conductivity instruments can be used for a wide range of concentrations by exchange of conductivity probes.
电导率仪器通过更换电导率探头，可以被用来测量很宽范围的浓度。
Conductivity readings are highly influenced by the sample temperature.
样品温度对电导率的读数有很大影响。
Either temperature adjustment of the sample or automated temperature compensation can be used to standardize the measurements.
调整样品温度或者自动温度补偿可以被用来标准化测量。
Conductivity is a nonspecific method that correlates linearly (within a defined range) to the ionconcentration in an aqueous sample.
电导率是一个相关于水样离子浓度的线性非特定方法。
Analytical instruments are robust and can be used on the manufacturing floor by trained personnel.分析仪器非常耐用，可以在生产现场被经过培训的人员使用。
The high influence of the sample temperature on the instrument reading should be considered to avoid incorrect results.
样品温度对仪器读数的重大影响必须被考虑，避免错误结果。
The method cannot differentiate between different ions.
这个方法不能因为是不同离子而不同。
Therefore, as for TOC, all conductivity results above the water baseline should be attributed to the contaminant in question (e.g., the cleaning agent).
因此，和 TOC 一样，所有电导率在水基准线上的结果应归因于污染（如：清洗剂）所致。
For biotechnology cleaning validation applications, conductivity is normally not used to detect product residues.
由于生物制品清洗验证的应用，电导率是通常不用于检测产品残留。
To allow correlation of conductivity readings with concentrations of cleaning agents, a dilutioncurve(conductivity vs. concentration) should be established (at a relevant temperature) by conductivitymeasurements of different dilutions in the relevant range near the acceptance value.
为允许电导率读数与清洗剂浓度的相关性，应该通过在接近可接受值的相关范围，测量不同稀释液的电导率，建立（在相关温度）稀释曲线（电导率 VS 浓度） 。
6.3.4 Visual Inspection
6.3.4 目视检查
Visual inspection is a qualitative method to determine cleanliness on specific equipment surfaces.
目视检查是一种检查设备表面是否清洁的等同方法
Visual inspection has been demonstrated to be a simple and effective direct sampling method in the evaluation of equipment cleanliness.
目视检查已被证实是衡量设备清洁的一种简单有效直接取样方法。
Visual inspection does have multiple weaknesses that are inherent.
目视检查也确实有很多内在的不足。
Extensive training and a detailed documented procedure are required to ensure that “visually clean” from one operator to the next is consistent.
为确保能从一个操作者到下一个操作者都持续的“目视清洁” ，大量的培训和一个详细的有记录的程序在是非常必要的。
What one can visually see will vary with distance, angle, lighting, the nature of the surface and inspector’s visual acuity.
目视结果会随着距离，角度，光线，表面本质和检查者的视觉敏锐度的变化而变化。
Some equipment surfaces (e.g., piping) are usually not accessible for visual inspection.
一些设备表面（如管道）通常不接受目视检查。
The use of optical equipment (e.g., mirrors, remote visual cameras or endoscopes) can help to facilitate visual inspection.
一些光学设备（如：镜子，远程可见摄像，内窥镜）的使用能帮助目视检查
In order to view some equipment areas, wear and tear on the equipment may occur (e.g., the disk stacks in a centrifuge are not designed to be removed and inspected for visual cleanliness after cleaning).
为了能看到某些设备的取样，可能损耗设备（如离心机里的圆盘不能被设计成是可移除的和请接收被目视检查）
In other cases, an operator may be required to enter a confined space for viewing equipment surfaces.
另一种情况，操作者可能需要进入一个限制的空间观察设备表面。
The visual inspection procedure should specify how operators are to deal with visual observations.
目视检查程序应详细记录操作者如何处理目视结果的。
Visual inspection could find four different types of visual observations: residue, surface anomalies, foreign object and water pooling.
目视检查可以找到中不同类的目视结果：残留，表面异常，异物，积水。
Residue is the main concern, which would constitute a visual failure when one is looking at theacceptability of a cleaning cycle.
残留是主要关注的，它可能导致目视失败，当评审清洁周期的可接受性的时候。
A sample of the residue should be collected for further testing, if possible, to assist in the investigation of the cause.
如果有必要，残留的样品应收集起来进行深入测试，有助于原因调查
Typically, surface anomalies and foreign objects are not considered visual inspection failures for cleaning validation purposes, but must be further investigated and corrected, as applicable.
典型的，表面异常和异物不认为以清洁验证为目的的目视检查失败是适当的，但必须进行深入调查和纠正，
Surface anomalies should be noted and a “suitability for use” assessment should be performed toremediate any issue(s) found.
表面异常应记录和应进行一个“适用性”评估去补救任何发现的问题。
Rouge is the most common type of surface anomaly discovered during visual inspection; rouge isgenerally considered a preventive maintenance problem, not a cleaning process problem.
Rouge 是目视检查中发现的非常普通的表面异物种类；rouge 通常被认为是一个预防性维护问题，不是清洗程序的问题。
Foreign objects and their removal should be noted. How the foreign object came to be in the equipment should also be investigated.
外来异物和其去除应注释。外来异物如何进入设备也应进行调查。
Water pooling should be documented, and the cause should be investigated.
积水应进行记录，其原因应进行调查。
All equipment surfaces that can be readily inspected visually should be visually inspected.
所有设备表面能容易被进行目视检查的应进行目视检查。
Visual inspection may not be performed on the interior of lines and tubing (although outlets may be inspected)on equipment where disassembly of the equipment is not practical or possible, or where inspection of the equipment could potentially be dangerous to the inspector (e.g., entry into a confined space).
目视检查不应用于设备管线和管道内部（虽然出口处可以被检查） ，拆解设备是不可行或者是不可能的，或者设备检查的地方可能对检查者有潜在的危险。
A visual inspection training program should be developed for visual inspection.
目视检查的目视检查培训程序应开发。
Inspectors typically should be trained and/or requalified on an established basis.
检查者应进行培训或者在建立已培训的基础上再确认
If visual inspection is not possible on an area of concern, it is important to ensure that other sampling methods (such as rinse sampling) can adequately detect potential residues of concern.
如果目视检查在相关的区域是不可能进行的，确保其他取样方法（如淋洗取样）能足以检测到潜在的相关残留是非常必要的。
The use of “visually clean” alone (in the absence of other analytical methods such as TOC or conductivity) is not generally used in the biotechnology industry, because all critical surfaces are not readily available for visual examination.
单独使用目视清洁（不使用其他分析方法如 TOC 或者电导率）在生物制药领域是不常用的。因为所有关键表面不溶液被目视检查。
6.4 Microbial Test
MethodsThe U.S. FDA’s cleaning validation guidance states that “Control of the bioburden through adequate cleaning and storage of equipment is important to ensure that subsequent sterilization or sanitization procedures achieve the necessary assurance of sterility.”
FDA 的清洁验证指南指出“通过对设备的充分清洁和存储来控制生物负载，对保证后续的灭菌或者消毒程序达到必要的无菌保证是非常重要的”
Endotoxin is a concern in that steam sterilization does not destroy or remove endotoxin.
内毒素是重要的，因为蒸汽灭菌不能消灭或者去除内毒素。
Thus, both bioburden and endotoxin are typically monitored and controlled during the manufacturing and cleaning processes.
因此，生物负载和内毒素在生产和清洁过程中，都被代表性的监测和控制。
Typically microbiology sampling is performed during all cleaning validation studies throughout the manufacturing process.
生产过程中的所有清洁验证研究必须进行代表性的生物学取样
6.4.1 EndotoxinTypically, endotoxin testing is performed for cleaning validation runs.
典型地，清洁验证需进行内毒素测试
Endotoxin testing may not need to be tested in upstream cell culture and initial purification processes where it is proven during process validation that one or more purification steps is able to effectively remove endotoxin that is present.
内毒素测试可不需要在上游细胞培养和初始纯化程序进行测试，上述程序被证明在工艺验证中一个或者更多纯化步骤可以影响内毒素去除效果。
Typically, a three logs or greater reduction of endotoxin in endotoxin removal steps is required to justify decisions not to test for endotoxin upstream.
在内毒素去除步骤，3 个对数单位或者更多内毒素的减少是必须的，这将是不测试上游内毒素决定因素。
Endotoxin methods are typically compendia methods.
内毒素方法是典型的药典方法
6.4.2 BioburdenTesting of bioburden is typically done through rinse water sampling, although other methods may be used.
淋洗水取样过程应完成生物负载测试，尽管其他方法可能被使用。
The benefit of rinse water sampling for bioburden is that it is convenient.
生物负载的淋洗水取样的优势是它非常方便。
Typically, rinse water sampling is being performed to verify removal of protein and cleaning agent(s), so one additional rinse water sample does not require significantly more work.
淋洗水取样被用来确认蛋白质和清洁剂的去除，所以一个额外的淋洗水样品不需要更多的工作。
Also, bioburden testing of rinse water is typically already a qualified method for testing water systems for bioburden.
淋洗水的生物负载测试是一个典型的，已经确的，测试水系统生物负载的方法。
The biggest weakness of rinse water sampling is that the full range of the acceptance criteria is not able to be utilized, for example,if 100 ml of rinse water is used for testing with an acceptance criterion of 10 CFU/mL
.淋洗水取样最大的缺点是：大范围的可接受标准不能被应用。比如：如果 100ml 淋洗水被用来测试，可接受标准是 10CFU/ml
The typical number of colonies that can be counted is 300 before TNTC (Too Numerous To Count) is achieved;
在获得 TNTC（太多数而不能数）之前，菌群的典型可数数目是 300。
this only allows an acceptance criterion of 3 CFU/mL before failing to meet the acceptance criteria.在满足可接受标准之前，只能允许 3CFU/ml 的可接受标准。
In most situations this is not an issue; it may result in the need to test smaller sample volumes.
在很多情况下这不是一个问题；这可能导致需要去检测更小容量的样品。
If this situation occurs, it is important to test an adequate amount of rinse to ensure that bioburden is detected.
如果这种情况发生，为保证生物负载被检测，足够数量的淋洗测试是非常重要的
Two methods for measuring directly on surfaces are swab and contact plate method.
测量直接表面的两种方法是擦拭法和接触碟法。
For swab samples, the swab can be desorbed, and a count can be made by a pour plate method.
擦拭样品，擦拭可以被吸附，可以被冲洗碟方法计数。
Contact plates are directly incubated and enumerated.
接触碟是直接培养和计数。
The biggest concern with contact plates and swab procedures is potentially exposing product contact surfaces to an unknown media or buffer solution from swabs;
接触碟和擦拭程序最大相关的是潜在暴露产品与未知培养基或者擦拭的缓冲溶液接触表面。Thus, acceptable removal of this media or buffer solution should be demonstrated before manufacturing can occur.
因此，这些培养基或者缓冲溶液的可接受移除应该在生产之前证明。
Another concern is that contact plates require flat surfaces.
另一个关注点是接触碟需要平整的表面。
文章来源：允咨生物GMP学苑
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