前Merck资深专家关于清洗验证限度的思考

2018-08-10 来源:bc体育 点击:667
文章来源:制药技术 38卷,第10  2018.08.10
摘要
Cleaning validation programs must have cleaning limits, worst-case residues to validate, and recovery factors to accurately determine how clean the equipment must be. If a program is to be robust, it must also address the question of which products should be validated, and how to test residue levels accurately, to assure compliance with the defined clean validation limits. This article presents an integrated approach that can establish the necessary information in an efficient, compliant manner.
清洗验证方案必须要有清洗限度、需验证的最差条件残留及回收率以准确确定设备必须要有多清洁。一个方案要有耐用性,则它必须说明应验证哪个产品,如何去准确测定残留水平,及如何确保符合设定的清洗验证限度。本文提供了一个可以以一种高效合规方式建立这些所需信息的整体方法。
 
An effective cleaning validation program requires substantial up-front work to withstand regulatory scrutiny. Acceptable residue limits (ARLs) must be defined prior to any cleaning validation and development work (1,2). The ARL is the level to which product residues must be removed to assure patient safety and that the subsequent product manufactured on the cleaned equipment will not be contaminated.
一个有效的清洗验证方案需要扎实的前期工作以能接受官方检查。在开展任何清洗验证和开发工作之前必须先建立可接受残留限度(ARLs)(1,2)。可接受残留限度(ARLs)指的是产品残留必须被去除到可以保证患者安全的水平,也是后续产品在清洁设备上生产不会被污染的水平。
A number of factors go into the determination of the ARL, including product dosage levels, batch sizes, and equipment product contact surface areas (3). The dosage and batch size parameters are typically well defined for a product before it gets to commercial production. The equipment product contact surface areas, however, require more planning and more steps to execute.
可接受残留限度(ARLs)的确定需要考虑一系列因素,包括产品剂量、批量及设备产品接触表面积(3)。一般来说,剂量和批量参数在产品进入商业化生产阶段以前就可以被确定的。然而,设备产品接触表面积可需要更多的规划和步骤来实现。
Many vendors do not supply product contact surface areas or even dimensions of the product contact surfaces. Measurements and calculations of each piece of equipment’s product contact surfaces must be completed, documented, and reviewed to determine the ARL of the product. Although it can be time- consuming to calculate the product contact surface area for a piece of equipment, the resulting figure only changes when the equipment is altered, which would only occur under a documented change control.
In addition to establishing a calculated ARL, the first criterion of a cleaning procedure is that the equipment be visibly clean after the cleaning process. The visible residue limit (VRL) is the level below which a residue is not visible, under defined viewing conditions. It can be a valuable tool when applied to a cleaning validation program. Viewing distance, viewing angle, and viewing light level must be defined for the facility and applied to the determination of a VRL.
许多供应商并不提供产品接触表面积或产品接触表面的尺寸。所以必须完成每一设备产品接触表面积的测量和计算,结果需记录并复核以确定产品的可接受残留限度(ARLs)。尽管计算每一设备的产品接触表面积是很费时的,不过该计算结果仅在设备发生改变时才会发生改变,设备的改变需遵循书面的变更控制程序。除了建立经计算的可接受残留限度(ARL),清洗程序的第一标准是设备经清洗后应是目视洁净的。目测残留限度(VRL)是指在规定的观察条件下残留低于此限度即不可见的水平。这是清洗验证方案中所运用的有用工具。在确定目测残留限度(VRL)时必须明确观察距离、观察角度和观察光照水平。
VRLs have been used for a number of cleaning validation applications (4,5). Once established, VRLs can be implemented for multiple aspects of a cleaning validation program (6), including routine confirmation that the equipment is cleaned to appropriate levels before changeover to a new product. One approach to cleaning validation is to validate every product manufactured at a site. This approach, however, is impractical at multi-product facilities.
目测残留限度(VRL)已被用于一些清洗验证中(4,5)。一经建立,目测残留限度(VRL)可被用于清洗验证的多个方面(6),包括将设备清洁至一定水平后用于另一产品生产前的日常确认。一种清洗验证的方法是对工厂内所生产的每个产品进行验证。然而,对于一个多产品设施,这一方法是不现实的。
To streamline validation efforts, products can be grouped together by therapeutic family or based on the hardest-to-clean products manufactured on the equipment. Defining the worst-case product can itself be a challenge.
为使验证工作简单化,可根据治疗类型将产品分组,或基于设备所生产的最难清洗产品。确定最难清洗产品本身也是一个挑战。
Solubility of the API is sometimes used as the criteria for determining “hardest to clean,” but this approach ignores the product excipients, which can often be more difficult to remove from equipment than the API.
有时候原料药的溶解度被用作确定“最难清洗”的标准,但这一方法忽略了产品辅料,辅料往往比原料药更难于从设备上去除。
Cleanability of the product residue more closely approximates the actual cleaning procedure necessary to result in acceptably clean equipment. Cleanability study conditions can range from simple immersion to vigorous cleaning actions (7-10). As long as the cleanability parameters are consistent, they can define a relative cleanability ranking of the site product residues.
产品残留的可清洗性越接近实际所需清洗程序,则越容易得到可接受的清洁设备。可清洗性研究条件可以简单的浸泡变化到剧烈的清洗操作 (7-10)。只要可清洗性参数是一致的,则可确定各个产品的相对可清洗性排序。
Even with an executed cleanability study, before validation studies can be executed, cleaning development studies must define and confirm the cleaning conditions necessary to clean product residue effectively, down to an acceptable level. Cleaning development studies define the critical cleaning parameters and their ranges, which will assure the consistent capabilities of the cleaning process. The critical cleaning parameters can include detergent definition and concentration, water temperature, cleaning action (e.g.,impingement, cascade, or manual scrubbing), contact time, and rinse times. The conditions established during cleaning development should be confirmed under an executed protocol prior to validation activities.
在进行验证研究之前所进行的可清洗性研究中,清洗开发研究必须确定并确认可有效清洗产品残留并降至可接受水平所需的清洗条件。清洗开发研究确定关键清洗参数及其范围,这将能确保清洗程序具有稳定的能力。关键清洗参数可以包括清洗剂的选择和浓度、水温、清洗操作(如冲洗、灌洗或人工擦洗)、接触时间和淋洗时间。应在验证活动前按照实施方案对清洗开发过程中所建立的清洗条件进行确认。
Finally, recovery factors must be established to demonstrate that swab or rinse samples taken after cleaning are representative of the cleanliness of the equipment (1, 2, and 11). To accomplish this, a known amount of the residue of interest is first spiked onto a coupon of the material of construction representing the manufacturing equipment. The spiked residue is recovered, either by using a swab wet with a solvent to dissolve the residue, or with a known volume of purified water to represent the final rinse of the cleaning procedure. The recovered samples are then tested to determine the amount of recovered residue. The recovery factor is the amount of residue recovered compared to the amount of residue spiked onto the coupon.
最后,必须建立回收率以证实清洗后所取的擦拭或淋洗样可以代表设备的清洁程度(1, 2, and 11)。为实现此目的,首先将已知量的目标残留物添加到可以代表生产设备材质的试件上。然后通过溶剂浸湿溶解残留物或通过已知量纯化水模拟清洗程序的最终淋洗进行添加残留的回收。最后对回收样品进行测试以确定残留物的回收量。回收率是指残留物的回收量与试件上残留物添加量的比值。
Although the data are typically generated for these cleaning validation parameters in separate efforts, a coordinated study can determine all of the factors using available data and an efficient set of experiments. This approach will save time and resources, and result in a more aligned set of cleaning limits, soils to be validated, and recovery factors for the cleaning validation studies.
尽管通常是单独获得这些清洗验证参数,但也可以采用已有数据及有效的实验组来确定所有这些参数。该方案将可以节约时间和资源,也为清洗验证研究形成更为统一的清洗限度、验证基础和回收率。
Acceptable residue limit determinations
可接受残留限度确定
The ARL must be determined prior to cleaning development, analytical method validation, visual limits and recovery factors. The subsequent data and recovery factors are all based relative to the ARL of the residue of interest. The primary goal of a health-based ARL is patient safety, and the recommended method to define a health-based calculation incorporates the acceptable daily exposure (ADE) (12), which is the amount of material one can ingest on a continued daily basis without harmful pharmacologic effect. Although the ADE approach is preferred as being more scientifically sound, the ADE can be compared to a currently used 1/1000th minimum-daily-dose (3)approach during the transition period to the use of ADEs to determine if additional validation work is necessary.
必须在清洗开发、分析方法验证、目测限度和回收率之前确定可接受残留限度(ARL)。后续数据和回收率均是基于相关目标残留物的可接受残留限度(ARL)。建立基于健康的ARL主要目的在于患者安全,建立基于健康计算所得限度的推荐方法是使用日可接受接触量(ADE),ADE是指一人在持续摄入的基础上可以摄入对人体不会产生有害药理作用的物料量。尽管ADE法更为科学合理而被推崇,在使用ADE法的过渡阶段可以将ADE法与目前在使用的千分之一最低日剂量法进行比较以确定是否需要进行额外的验证工作。
The factors that go into an ARL calculation include the dose and the batch size of the next product, which determine the degree to which any carryover is spread among subsequent product dosages. The product contact surface area of the manufacturing equipment assumes an even distribution of any worst- case carry over residue. This assumption of even residue distribution is addressed by swabbing worst-case, hardest-to-clean and critical equipment locations; that is, those locations where residue is most likely to build up, or where residue could be transferred to a small number of subsequent doses.
The swab area and the recovery factor enable one to relate the result for a single sample to a total residue amount in the manufacturing equipment. The health-based calculation employing the ADE is shown in Equation 1:
ARL计算需要考虑的因素包括下一产品的剂量和批量,这些决定着残留物在下一产品中的残留程度。假设生产设备产品接触表面均匀分布了最差条件的残留物。该残留物均匀分布的假设在基于擦取了最差条件、最难清洗和最关键的设备位置,即那些残留物最有可能富集,或可能会被携带至后续产品去的位置。擦拭面积 和回收率可以使得单个样品的结果能与生产设备上的总体残留量建立相关性。运用ADE进行基于健康的ARL计算公式如式1所示:
(ADE (mg/day))/(MDD (doses/day)) x (BS (doses))/(SA (cm2)) xM (cm2/swab)x RF=ARL (mg/swab) (Eq. 1)
式1:(ADE (毫克/天))/(MDD (剂量/天)) x (BS (剂量))/(SA (厘米 2)) xM (厘米 2/擦拭)x RF=ARL (毫克/擦拭)
Where:
其中:
ADE is ADE of Product A being cleaned (in mg/day)
ADE指的是所清洗产品A的ADE(毫克/天)
MDD is maximum daily dose of subsequent Product B (in doses/day)
MDD指的是后续产品B(剂量/天)的最大日剂量;
BS is batch size of product B (in number of doses)
BS是产品B的批量(剂量的数量)
SA is product contact surface area of the equipment train (in cm2)
SA是设备组的产品接触表面积(厘米 2)
M is swab area = 25cm2
M是擦拭面积=25厘米 2
RF is recovery factor (e.g., 0.90 for a 90% Recovery Factor)
RF是回收率(如,0.90指的是90%回收率)
ARL is acceptable residue limit.
ARL是可接受残留限度。
 
The intent is to consider the health-based ARL and the VRL to satisfy regulatory requirements for cleaning so that the patient is safe and the equipment is visually clean. Although the VRL for residues should be related to the health- based cleaning limit, as well as the analytical detection limit (LOD), the VRLs can be determined prior to defining the final ARL because the VRL is more an experimentally determined physical characteristic of the API or product established under defined condition rather than a relative characteristic based on other factors.
考虑基于健康的ARL和VRL的目的在于满足清洗法规要求,使得患者安全和设备洁净。尽管残留的VRL应与基于健康的清洗限度相关,也与分析检出限(LOD)有关。但可在确定最终ARL之前建立VRL,因为VRL是更为由实验在指定实验条件下测定的原料药或产品物理属性,而不是基于其它因素的相对属性。
Visible residue limit studies
目测残留限度(VRL)研究
VRLs must be determined using well-defined viewing parameters to better transfer the implementation of the VRLs to the production equipment and to limit subjectivity. The viewing variables associated with studying visible residue must be defined, and then experimental parameters for the study can be established. The parameters considered are:
必须使用明确的观察参数来建立VRL以能更好地将VRL运用到生产设备上,也能减少主观性。必须明确与研究目测残留研究相关的观察变量,然后才能建立研究实验参数。所需考虑的参数包括:
Equipment material of construction
设备材质
Light intensity
光强度
Viewing distance
观察距离
Viewing angle
观察角度
Observer subjectivity
观察者的主观性
 Solvent effects.
溶剂影响
Stainless steel is an obvious choice for surface material, because more than 95% of manufacturing equipment surfaces at a typical pharmaceutical manufacturing site are made from this material. Representative stainless-steel coupons are used for spotting purposes in the laboratory setting.
In addition to stainless steel, other widely used materials of construction (MOC) include:
由于在一家典型的制药厂超过95%的生产表面是不锈钢材质的,显然不锈钢是表面材质的一种选择。因此会采用有代表性不锈钢试件进行加样试验。除了不锈钢之外,其它广为使用的材质包括:
Polytetrafluoroethylene (PTFE), including Teflon
聚四氟乙烯(PTFE),包括特氟龙
High-density polyethylene (HDPE)
高密度聚乙烯(HDPE)
Low-density polyethylene (LDPE)
低密度聚乙烯(LDPE)
Polycarbonates, including Lexan
聚碳酸酯,包括莱克森
Glass, which can be addressed as part of the cleanability determinations.
玻璃,将作为可清洗性测定的一部分进行阐述。
Although the VRLs for Lexan and glass are comparable to that of stainless steel (13), VRLs for the remaining MOCs would be higher than the VRL for stainless steel. Cleanability provides a much simpler answer to the question of VRLs for different MOCs.
尽管莱克森和玻璃的目测残留限度(VRL)与不锈钢的相当,但其它一些材质的VRL会较不锈钢的高。关于不同材质VRL的问题,可清洗性给出了简单很多的答案。
Lighting conditions in the manufacturing plant typically differ, from room to room. The light intensity is measured in each room of the plant and the wash area to determine a range. For consistency, the light measurement is taken in the same location in each room, for example in the center of each room, approximately four feet from the ground. The light level in a typical pharmaceutical manufacturing plant generally ranges from 200-1000 lux.
一般来说,生产厂区不同房间内的光照条件会不同。需在工厂的每个房间和清洗区域测量光强度以确定范围。为保持一致性,应在每个房间的相同位置进行光强度的测定,比如在每个房间的中间,离地约4英尺的地方。在典型的制药厂内,光照水平范围一般为200~1000勒克斯。
The viewing distance and viewing angle are based on the manufacturing equipment that is used at the site. Larger pieces of equipment can often be viewed at a distance of no greater than 10 feet, and, if the equipment is disassembled for cleaning, the viewing angles are marginally limited for visual inspections.
Suspensions of the products are prepared in methanol at concentrations of the API and spiked onto stainless-steel coupons. For products compressed from common formulation blends, the highest potency product is used for the VRL determination. For the remaining products, the single strength manufactured at the site is used for VRL determination.
观察距离和观察角度取决于现场所用的生产设备。较大设备通常可以在不超过10英尺的距离内进行观察,如果设备是拆卸清洗的,则观察角度对于目视检查限度较少。配制产品的甲醇悬浮液,以原料药计算浓度,然后添加到不锈钢试件上。对于共用混粉设备的产品,应选择药效最强的产品进行VRL测定。对于其它产品,采用单一剂量进行VRL测定。
The spiked coupons are allowed to air dry, and the distance, angle and light level viewing parameters are set. Site personnel view the spiked coupons from multiple distances and angles. The VRLs are determined at a distance of two feet. Increasing viewing distance and viewing angle observations of the spiked coupons establish the viewing parameter limitations on the ability to see the VRL levels. If the observers are not able to see the VRLs at the distance of the larger equipment, this limits the use of VRLs to those pieces of equipment that can be viewed from established VRL viewing limitations. Literature references (6) have shown that most VRLs can be detected from 10 feet. Any seeming inconsistency is likely a result of very low VRLs that were determined experimentally. Some VRLs can be detected at levels as low as 0.1?g/cm2, compared to the literature average of 1.1?g/cm2. The viewing angle should be greater than 30 and the light level should be greater than 200 lux (6).
将加样试件晾干,设定好距离、角度和光照强度等观察参数。工厂人员从多个距离和角度观察加样试件。在两英尺的距离处测定VRL。提高加样试件的观察距离和观察角度可以建立观察VRL水平之能力的观察参数限度。如果观察者不能在较大设备的距离观察到VRL,则这将限制VRL运用于那些能从已确定VRL观察限度处所观察的设备。参考文献(6) 表明绝大多数VRL可以在10英尺处被观察。任何貌似不一致的情况都可能是由于实验测定的VRL非常低。相比于文献中平均值1.1微克/平方厘米,有些VRL可以低至0.1微克/平方厘米。观察角度应大于30,光照水平应大于200勒克斯 (6)。
Table I: Spiking preparation and target concentrations.
表I: 加样配制和目标浓度
 
The ARL and VRL will indicate how clean the equipment needs to be. Table I shows results spiking preparation and target concentrations for one test. The next step is to determine which product(s) or residue(s) to validate.
ARL和VRL将表明设备所需达到的洁净程度。表I表明每项检测的加样配制和目标浓度。下一次确定对哪个产品或残留进行验证。
Cleanability/cleaning development studies
可清洗性/清洗开发研究
For efficiency and economy, the initial cleaning development work can be conducted at laboratory scale in three phases. These laboratory-scale process and cleaner studies (PACE evaluation) were executed in the technical laboratory at STERIS Corporation, St. Louis, Missouri. The Phase 1 studies challenge a worst-case set of conditions. Baked-on residues are cleaned using different cleaning agents, cleaning mechanisms, times and temperatures. Once the optimal cleaning agent and conditions have been identified, the cleaning parameters are challenged in Phase 2 studies, to determine the minimum times and concentrations necessary to achieve clean equipment. In the Phase 3 study, the minimum cleaning parameters identified in Phase 2 studies are used to clean the worst-case soil identified in Phase 1 studies, from the MOCs that make up most of the product contact surfaces of the equipment at the facility.
In Phase 1 of the study, to create worst-case conditions, cleanability studies are performed on the products and blends manufactured at the facility, to determine which cleaning agent will adequately remove product residue. The study results will provide cleaning conditions, including the concentration of detergent to be used, critical cleaning parameters, time for rinse, etc.
考虑到效率和经济,可以在实验室规模进行最初的清洗开发工作,包括三个阶段。这些实验室规模的工艺和清洁剂研究(PACE评估)是在美国密苏里州圣路易斯市STERIS公司的技术实验室完成的。第1阶段研究是对最差条件组合进行挑战。采取不同的清洗剂、清洗机理、次数和温度对炼制残留进行清洗。第2阶段研究对清洗参数进行挑战,以确定可获得清洁设备的最少次数和最低浓度。第3阶段研究中,采用第2阶段研究所识别最低清洗参数将第1阶段研究所识别的最差情况物料从工厂 绝大部分设备产品接触表面所用材质上清洗掉。在第1阶段研究中,为创造最差条件,可清洗性研究是采用工厂内所生产的产品和混合物进行研究的,以确定哪种清洗剂可以充分去除产品残留。研究结果将提供清洁条件,包括所用清洁剂的浓度,关键清洗参数和淋洗时间等。
In the Phase I cleanability study, dry, clean stainless-steel coupons are weighed on an analytical balance (±0.1 mg) to obtain their pre-coating weight; then they are coated with 3-5 mL of 10% w/v slurry or 3-5 grams of sample. They are then baked at 57 °C for 4 hours then air dried overnight, and weighed on an analytical balance. The coated surface area is measured, the dry coating weight calculated, and the “loading” of the sample, in milligrams per square centimeter of dried residue, is determined. The spiked coupons can be cleaned by agitated immersion, spray wash (11 psi), cascading flow, or scrubbed manually using a nylon-bristled brush; in addition to the cleaning technique, the type and concentration of detergent, the cleaning temperature, and the cleaning time are recorded.
在第1阶段可清洗性研究中,先将干的洁净不锈钢试件在分析天平(±0.1 毫克)上进行称量得到加样前的重量,然后会将其涂上3-5毫升10%W/V浆状物料或3-5克样品。然后将其在57 °C烘制4小时后风干过夜,再在分析天平上称重。测量涂料表面积,计算涂料干重,则可计算出每平方厘米上所“装载”的样品重量(毫克/平方厘米)。通过搅拌浸泡、淋洗(11psi),冲洗,或采用尼龙刚毛刷进行人工擦洗。除了清洗程序之外,还需记录清洗剂的类型的浓度、清洗温度和清洗时间。
After cleaning, the coupons are removed and visually observed for cleanliness; then, each side of the coupon is rinsed, first, with tap water for 10 seconds at a flow rate of 0.5 gal/min and then with de-ionized water. It is then examined for a water break-free (WBF) surface, after which it is dried and weighed on an analytical balance to determine the post-cleaning weight.
清洗后,取出试件目测其洁净程度,然后以0.5加仑/分的速度采用自来水淋洗试件的两面10秒钟,之后再采用去离子水清洗。然后再检查表面无水膜残留,再经干燥后在分析天平上称其清洗后重量。
Table II: Phase 1 PACE study results (CIP is clean in place).
表II:第1阶段PACE研究结果(CIP指的是在线清洗)
 
A coupon is considered to be clean if it is visually clean, water break-free, and if its pre-coating weight and post-cleaning weight are equal (0.0 mg residue).
如果试件目测洁净,无水膜残留,且其加样前重量和清洗后重量是相等的(0.0mg残留),则试样可被认为是洁净的。
WBF is a qualitative test that indicates the cleanliness of a metal surface. On a clean surface, free from organic residue, water sheets evenly without breaks in the water film as it runs from the surface of the metal panel. The results of a Phase 1 case study are shown in Table II, with the worst-case soils designated. Two soils are designated as worst-case Product A is manufactured in dedicated equipment. Product B is manufactured in multi-use equipment with the remainder of the site product portfolio.
无水膜残留是项定性测试,表明金属表面的洁净度。在一洁净表面,若无有机物残留,则水在经过金属板表面时会很均匀而无水膜破裂。表II中为第1阶段采用最差条件物料的研究结果。选取了两种最差条件物料,产品A是在专用设备上生产的。产品B是多用途设备上生产的,设备会有产品组合的残留。
Phase 2 study
第2阶段研究
Phase 2 studies further evaluate the effectiveness of the cleaning agents that demonstrated positive results in Phase 1. These tests are run under conditions that more realistically reflect what would be experienced in actual use. The major difference in the Phase 2 coupon preparation is that the coupons are air dried only for 24 hours, rather than dried in an oven. Phase 2 cleanability studies are performed to provide a more focused look at critical cleaning process parameters such as time, temperature, concentration of detergent, and cleaning agent, and to demonstrate the ruggedness of the cleaning parameters identified in Phase 1 studies. The two worst cases, one product and one blend, from Phase 1 testing, are tested in Phase 2 to minimize testing resources and the results shown in Table III.
第2阶段研究进一步评估了清洗剂的有效性以阐述第1阶段的阳性结果。这些实验是在更为实际的条件下开展的,反映了实际使用过程中会经历的情况。第2阶段试件制作的主要区别在于试件仅是风干24小时,而不是在烘箱中烘制。第2阶段的可清洗性研究更聚焦于关键清洗工艺参数,如时间、温度、清洗剂浓度,和清洗剂以阐述第1阶段所识别的清洗参数的耐用性。第2阶段研究中对于第1阶段所选择的两种最差条件下的一个产品和一个混合物进行了检测以减少检测资源,研究结果见表III。
Table III: Phase 2 PACE study results.
表III:第2阶段PACE研究结果
 
 
Phase 3 study
第3阶段研究
In Phase 3 studies, the minimum cleaning parameters identified in Phase 2 studies are used to clean the worst-case soil identified in Phase 1 studies from the MOCs that make up most of the product contact surfaces of the equipment at the facility. The worst-case product is applied to different MOC coupons, air dried for 24 hours, and cleaned using agitated immersion at the previously identified cleaning parameters. The results are shown in Table IV.
在第3阶段研究中,按照第2阶段研究所识别的最低清洗参数条件对第1阶段研究所识别的最差物料在工厂所用具有主要产品接触表面的设备材质上进行了研究。将最差物料加至不同材质的试件上,风干24小时,然后按照之前所识别的清洗参数进行搅拌浸泡。结果详见表IV。
Table IV: Phase 3 PACE study results-Product B blend. (CIP is clean in place. HDPE is high- density polyethylene and LDPE is low-density polyethylene.)
表IV:第3阶段PACE研究结果-混合产品B(CIP指的是在线清洗,HDPE指的高密度聚乙烯,LDPE指的是低密度聚乙烯)
 
If all of the materials of construction are cleaned for the worst-case soil under the same cleaning conditions, it can be concluded that, if the equipment’s stainless-steel surfaces are clean, the other equipment surfaces are cleaned to the same level of cleanliness. An acceptable visual inspection of the stainless- steel surfaces would provide confidence that the other surfaces such as PTFE, HDPE, or LDPE, on which spots would be more difficult to detect, are clean to the same acceptable level.
如果所有材质均是在同样的清洗条件下去除最差条件物料,则可以推断出若设备的不锈钢表面是清洁的,则其它设备表面也可以被清洗至同样的清洁度。不锈钢表面的可接受目视检查会对其它表面的目视检查达到同样的可接受水平提供信心,如PTFE、HDPE和LDPE,在这些材质上会更难去发现残留。
Swab recovery studies
擦拭回收研究
Swab sampling is the preferred technique to determine equipment cleanliness because it is direct surface sampling and targets hard-to-clean and critical locations such as tablet press tooling.Swab sampling is generally more sensitive than rinse sampling because of the larger volumes associated with final rinses. An accurate swab sample requires that a swab recovery factor be established. The swab recovery factor is established by spiking a known amount of the API or product formulation onto a material of construction coupon, letting it dry, and swabbing the coupon to recover the residue.
擦拭取样是确定设备清洁度的推崇技术,因为它是直接表面取样,也可以针对于难清洗的关键部位,如压片机部件。相比于淋洗取样,擦拭取样通常更具灵敏性,因为最后淋洗的体积大。正确的擦拭样品需要建立擦拭回收率。擦拭回收率是通过将已知量的原料药或药品加至试件上,经风干后擦拭回收残留建立的。
To execute a swab recovery study, the parameters of the study must first be defined. These parameters include the coupon MOC, swab area, swab manufacturer and mode, number of swabs, swab solvent, swab technique, the extraction solvent, and the swab extraction parameters. Once these parameters are defined, they can often be applied across the APIs and products that require recovery factors. The last remaining parameter is the level of analyte to recover (i.e., the amount of analyte to spike on the coupons).
为开展擦拭回收研究,需首先确定研究参数。这些参数包括试件材质、擦拭面积、棉签厂家和类型、棉签数量、擦拭溶剂、擦拭技术、提取溶剂和擦拭提取参数。一旦这些参数确定了,则可运用于需要回收参数的原料药和药品。最后一个参数就是待回收分析物的水平(即,加在试件上待分析物的量)。
The logical level at which to perform a recovery is at the cleaning limit itself, because the cleaning limit is the pass/fail point of the residue test. For relatively safe products, the health-based cleaning limits are often quite high, for example greater than 1 to 10 mg/swab, which most likely would overload the swab and result in low recoveries. These levels also would be easily visible, which would fail the cleaning before the swabs are even taken. An efficient level for recoveries would be around the VRL level, because samples were already made for the VRL determination. For example, samples can be prepared at 5.0, 7.5, 10, and 12.5 μg of API/swab, slightly higher than the reported average VRL of 1.1 μg/cm2 .
These levels are also close to the levels that one would expect to see after cleaning. The risk is that these levels are also close to the limit of quantitation (LOQ) of the analytical test method and could result in low recoveries with high % relative standard deviations (RSDs).
进行回收研究时符合逻辑的水平是清洗限度本身,因为清洗限度是残留检测的通过/失败点。对于相对安全的产品,其基于健康的清洗限度往往相当高,比如会高于10毫克/擦拭,这很有可能会过载从而导致低回收率。这样的水平也是很容易被目测的,这意味着在进行擦拭前清洗就已经失败了。有效的回收水平应是在VRL水平左右,因为样品是已经经过VRL检测的。比如,可以配制5.0、7.0、10和12.5微克原料药/擦拭,略高于所报导的平均VRL,1.1微克/平方厘米。这些水平是与清洗后所期望的水平相接近的。这些水平的风险在于较分析方法的定量限(LOQ)较近,会导致出现高相对标准偏差(RSD)的低回收率。
The swab recovery samples are prepared using the suspensions prepared for the VRL study. The 5.0-, 7.5-, 10-, and 12.5-μg samples are spiked onto stainless-steel coupons using 50, 75, 100, and 125 μL, respectively, of the 100- μg/mL suspensions. The spiked coupons are allowed to dry. For swabbing, each sample container is labeled, recording the API/product, amount (?L), volume (mL), name, and date.
The swab is wetted with methanol, water, or other appropriate solvent, which will dissolve the API. Any excess solvent is removed by pressing the swab against the side of the sample container to wring out excess solvent from the tip. The swab area is at least 25 cm2, and pains must be taken to ensure that all the area covered by the dried residue is swabbed.
采用VRL研究时配制的悬浮液配制擦拭回收样品。分别通过添加50、75、100和125微升100微克/毫升悬浮液将5.0、7.5、10和12.5微克样品加至不锈钢试件上。将加样后的试件晾干。每个样品盒需加上标签,记录原料药/药品、量(微升)、体积(毫升)、姓名和日期。使用甲醇、水或其它可以溶解原料药的适宜溶剂润湿棉签。通过将棉签压在样品盒的侧面去除棉签头上多余的溶剂。擦拭面积至少为25平方厘米,必须要承受的擦拭痛苦在于确保所有被残留物附着的区域均需被擦拭。
The swabbing technique is shown in Figure 1. Using the flat side of the swab, slight pressure is applied to the swab stick, and full contact is made with the coupon. The coupon should be swabbed, using a back-and-forth motion, for approximately 10 seconds. Then, one should flip the swab over and swab the coupon in a perpendicular direction, using a back-and-forth motion, for approximately 10 seconds. Finally, one should snap the swab head into the sample container and close the container. The swabbing is repeated for each coupon and the samples submitted for analysis. The validated high- performance liquid chromatography (HPLC) test methods used for analysis should be specific for each analyte.
擦拭技术详见图1。使用棉签扁端,在棉签杆上略施压力以使其充分接触试件。采用前后移动的方式擦拭试件约10秒钟。然后旋转棉签再从垂直方向前后移动方式擦拭约10秒钟。对每一试件采用相同的擦拭方法进行擦拭,然后送样分析。所使用的经验证的HPLC方法需对每一个分析物是有专属性的。
Figure 1: Swab sampling technique.
图1:擦拭取样技术
 
The swab recovery results should be greater than 70% for stainless steel based on historic data (14). The variability (%RSD) should be less than 10%. However, performing recoveries at levels close to the analytical method LOQ can result in lower-than-expected recoveries, with higher %RSDs. The results will be affected by a number of factors:
根据历史数据不锈钢的的擦拭回收率应大于70% (14)。变化幅度(%RSD)应小于10%。不过,在接近分析方法定量限(LOQ)的水平进行回收时会导致出现低于预期的回收率,且变化幅度(%RSD)高。如下因素会影响回收结果:
Low spike levels near the LOQ of the analytical methods
接近分析方法定量限的低加样水平
Experience level of the staff performing the recoveries
从事回收研究人员的熟练程度
Robustness of the extraction parameters
提取参数的耐用性
Size of the swab head.
棉签头的大小
A larger swab head would be expected to retain slightly more residue than a smaller swab head of the same material. Ideally, the swab recovery spike levels are around the VRL, but these levels are likely to be too low for quantitation by the HPLC methods after swab recovery and extraction. That is why the swab spike levels are targeted slightly above the method LOQs. The proximity of the spike levels and the method LOQs could contribute to both low recoveries and high variability if small, variable amounts of analyte adhere to the swab. Also, the lower HPLC area counts near the LOQ could contribute to the higher %RSD compared to comparably spread data with greater HPLC area counts.
相比于较小的棉签头,同一材质的较大棉签头可以保持更多的残留物。理想情况下,擦拭回收加样水平应在VRL附近,但这些水平很有可能对于擦拭回收和提取后采用HPLC方法进行定量检测时水平太低了。
Typical validated HPLC methods have LOQs of approximately 1 ?g/mL. If the methods can be optimized with real samples prior to validation, a lower LOQ can often be established, and better recovery conditions identified. The effort required for optimization, however, may not be worth marginal improvements to the recovery data, especially when compared to the calculated ADE-based cleaning limit. Although the personnel involved in the recoveries are a factor, their contribution to low and variable results is considered minor compared to analytical issues (14).
经验证的典型HPLC方法的定量限(LOQ)约为1微克/毫升。如果在验证前可以采用实际样品进行分析方法优化,则常常可以得到更低的定量限(LOQ),也可以识别更好的回收条件。不过,相比于回收数据的略微改进,为获得方法改进所作努力可能是不值得的,特别是与基于ADE所计算的清洗限度相比较时。尽管参与回收的人员也是一个影响因素,相比于分析影响,他们的影响是对于低而变化的回收结果而言是小的(14)。
Rinse-recovery studies
淋洗-回收研究
Rinse-recovery studies are conducted using the materials and previously prepared solutions or suspensions to offer the flexibility to use rinse sampling. Although rinse sampling is considered indirect surface sampling, it covers all of the product contact surface area and more easily samples those areas that are inaccessible to swab sampling. In addition, rinse samples are easier to take and more efficient to test. Rinse sampling, however, is generally less sensitive than swab sampling because of the larger volumes associated with final rinses. The rinse-recovery factor is established by spiking a known amount of the API or product onto a material of construction coupon, letting it dry, and rinsing the coupon to recover the residue.
可以采用物料、之前配制的溶液或悬浮液进行淋洗-回收研究以为淋洗取样提供灵活性。尽管淋洗取样被认为是间接表面取样,但它涵盖了所有的产品接触表面区域,并可以对不能进行擦拭取样的区域进行更为容易的取样。此外,淋洗取样更易进行,检测也更为有效。不过,淋洗取样通常被认为不如擦拭取样更具灵敏度,因为最后的淋洗有较大的体积。淋洗回收率是通过将已知量的原料药或产品加至试件上,晾干,然后再淋洗试件回收残留建立的。
To execute a rinse-recovery study, the parameters of the study must first be defined. These parameters include the level of analyte to recover, the coupon MOC, the rinse area, the rinse solvent, and the rinse volume. Once these parameters have been defined, they can often be applied across the APIs and products that require recovery factors.
在开展淋洗-回收研究时,必须先确定研究参数。这些参数包括待回收分析物的水平、试件材质、淋洗面积、淋洗溶剂、和淋洗体积。一旦确定了这些参数,它们常常可以被运用于需要回收参数的原料药和药品中。
To be consistent with the swab recoveries, the spike level for the rinse recoveries can be set at 10 ?g of API or product onto a 25-cm2 area of stainless-steel coupon. Final rinses are all done with purified water, and a volume of 10-25 mL is used, making sure to keep the final solution at or above the method LOQ.
The rinse recovery results should be more than 70% for stainless steel. The variability (%RSD) should be less than 10%. However, performing rinse recoveries at levels close to the analytical method LOQ can result in lower than expected recoveries and with high %RSDs. The results will be affected by a number of factors: the low recovery levels near the LOQ of the analytical methods and the volume of the rinse water. Too small a volume will not remove the residue and too large a volume will be undetectable. For the residues with high %RSD as well as those for which no quantitative values, an investigation should be conducted to determine a root cause.
为与擦拭回收保持一致,淋洗回收的加样水平可设为10微克原料药或药品,将其加在25平方厘米的不锈钢试件上。采用纯化水进行最后的淋洗,所用体积为10-25毫升,确保最后的溶液浓度处于或高于分析方法定量限(LOQ)。对于不锈钢,淋洗回收率应超过70%。偏差(%RSD)应小于10%。不过,在接近于分析方法定量限的水平进行淋洗回收研究会导致低于预期的回收率,且偏差高。回收率会受一些因素影响:接近分析方法定量限的低回收水平和淋洗液的体积。太小体积不能去除残留物,而太大体积而不能被检出。对于高偏差(%RSD)的残留及那些没有定量值的残留,应进行调查以确定根本原因。
Discussion
讨论
Although the described studies serve to establish the necessary background data for the cleaning validation effort, there were several issues that could be addressed differently. The VRLs are established under laboratory conditions, and recent VRL levels obtained averaged 0.1 ?g/cm2. This raises the concern that data might not translate to full-size equipment in the manufacturing plant. Future work would answer that question, by taking spiked coupons and placing them inside actual equipment, or equivalent conditions, to confirm the laboratory generated data.
尽管所阐述的研究可以为清洗验证工作建立必要的背景信息,有些问题还需另作阐述。目测残留水平(VRL)是在实验室条件建立的,最近获得的VRL水平平均为0.1微克/平方厘米。这会存在该数据是否能在生产工厂的实际设备上实现的顾虑。未来的工作将回答该问题,通过将加样试件摆入实际设备,或等同条件,可证实实验室所产生的数据。
Very low VRLs also raise the question of how the VRLs are defined. Past work had defined the VRL concentration by dividing the amount of material spiked by the entire surface area of the circle formed, even though most of the material forms a ring, leaving the middle of the circle empty. This is not a concern as long as the VRL determinations have been defined consistently. With the VRL defined using this approach, however, the VRL of residues could approach or even be lower than the LOQ of the analytical method. Defining the VRL as the area of only the ring and not the entire circle might be a better, more realistic approach, and could relate more closely to the recovery factors and the analytical test method LOQ.
非常低的VRL也会引发VRL是如何确定的疑问。过去的工作已经确定了VRL浓度,方式是将所加物料的是除以所形成圆形的面积,即使大部分物料形成了一个环,而圆中间是空的。只要VRL的测定是一致的,则这并不成为问题。然而,对于通过该方法确定的VRL,处于VRL水平的残留会接近甚至低于分析方法的定量限。通过选择仅是环的面积而不是整个圆形的面积来计算VRL可能会更好,也是更为真实的方法,这将与回收率和分析方法定量限更为接近。
The LOQ of the HPLC analytical test methods should be optimized for cleaning validation samples. A lower LOQ would alleviate some of the concern with the VRL levels and probably would improve the variability of the swab and rinse recovery studies. The added value must be deemed significant enough, however, to expend the additional resources for this work. Coordination with the testing laboratories is essential during cleaning development work.
对于清洗验证样品,应对HPLC分析方法的定量限进行优化。较低的定量限将减少关于VRL水平的顾虑,也可能会改善擦拭和淋洗回收研究的偏差。为本工作投入额外的资源应被视为是有显著价值的。在清洗开发工作过程上有必须与检测实验室进行协作。
The rinse-recovery study levels of 10 ?g are based on swab recovery levels. Because the rinse volumes are higher than the swab extraction volumes, the resulting rinse-recovery concentrations are lower. To ensure accurate data, the rinse recovery samples should be spiked based on the final concentration of solution (?g/mL) of the rinse-recoveries.
将淋洗回收研究水平设为10微克是其于擦拭回收水平而设定的。因为淋洗体积高于擦拭提取体积,导致淋洗回收浓度较低。为保证数据的准确,淋洗回收样品的加入应基于淋洗回收溶液的最终浓度(毫克/毫升)。
Conclusion
结论
Background data for a cleaning validation program can be generated in an efficient, coordinated effort for a pharmaceutical manufacturing facility. Using this approach, the ARL calculations and the cleanability data are combined to define the worst-case product for cleaning. The VRL, swab recoveries, and rinse recoveries are established using a single set of product suspensions, which illustrate the relationship among the three factors. These studies clearly demonstrate that a cleaning validation program can be established or revalidated using an efficient, coordinated effort to establish the necessary background information.
在一个药品生产企业,可以通过高效协调工作获得清洗验证所需的背景信息。使用该方法,可以结合可接受残留限度(ARL)计算和可清洗性数据确定清洗的最差产品。采用一组产品悬浮液建立了目测残留限度(VRL)、擦拭回收率和淋洗回收率,并阐述了三者之间的关系。这些研究清楚地阐述了可以采用高效协作地方式获得必要的背景信息以进行清洗验证或再验证。

作者:理查德 J.福赛思[1]

制药技术 38卷,第10


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