1、質保處質保處2009.05 過濾知識及原理過濾知識及原理 過濾器的使用及維護過濾器的使用及維護 完整性測試完整性測試 自動完整性測試儀自動完整性測試儀 Important differences in the types of filters used in Bioprocesses. 生物制藥中使用的不同類型過濾器及生物制藥中使用的不同類型過濾器及其特性其特性 Why we use membrane filters for critical application 關鍵應用中為什么使用膜過濾器關鍵應用中為什么使用膜過濾器 How to recognize filtration problem

2、s and perform basictroubleshooting 怎樣理解過濾中的問題和解決簡單故障怎樣理解過濾中的問題和解決簡單故障 Size Exclusion大小排除大小排除 Particles are too large to pass through the “pore” 基于大小來分離基于大小來分離 Examples are termed entrapment,sieving, surface capture 例如攔截例如攔截,篩分篩分,表面捕獲表面捕獲 Adsorption吸附吸附 Particles stick to the filter bychemical / elec

3、trical attraction 顆粒由于化學顆粒由于化學/電荷作用粘附在濾器上電荷作用粘附在濾器上 Mechanisms can be affected by: 截流機制可以被以下因素影響：截流機制可以被以下因素影響： Fluid characteristics 流體特性流體特性 Operating conditions 操作條件操作條件 Particle type 顆粒類型顆粒類型 Filter type 過濾膜類型過濾膜類型 Viscosity粘度粘度 Chemical / Ionic content 化學化學/離子成分離子成分 Comments Slow flow allows mo

4、re contact較慢流速有較多接觸較慢流速有較多接觸 Mixing/Contact time can be critical混合混合/接觸時間有較大影接觸時間有較大影響響 Only affect adsorptive retention 只影響吸附截留只影響吸附截留 - NOT size exclusion 不影響大小排除不影響大小排除 Particle speed / velocity 顆粒速度顆粒速度 Applied pressure過濾壓力過濾壓力 Comments建議建議 Slow flow is good選擇低流速選擇低流速 better retention at slower

5、flow越低的流速越低的流速,越好的截留越好的截留 Matrix movement is bad結構移動是不利的結構移動是不利的 particles and fibers can bereleased from depth filters 顆粒和纖維能從深層過濾器析出顆粒和纖維能從深層過濾器析出 Velocity/Differential Pressure can have a major effect on adsorptive retention - Little to no effect on size exclusion 速度速度/壓差對吸附截留有壓差對吸附截留有重要影響重要影響,對大

6、小排除影響相當小對大小排除影響相當小 Deformable (soft)可變形顆粒可變形顆粒 Non-deformable (hard)不可變形顆粒不可變形顆粒 Comments建議建議 Under pressure soft particles canintrude into the filter structurecausing more effective filter blinding 在壓力下軟性顆粒會進入過濾膜內并導致更多的過濾堵在壓力下軟性顆粒會進入過濾膜內并導致更多的過濾堵塞塞 例如例如:凝膠凝膠 Hard particles form open cakes 硬顆粒形成一層餅例

7、如硬顆粒形成一層餅例如:沙子沙子 Pore rating孔徑孔徑 Structure結構結構 Rigid or movable剛性或可移動剛性或可移動 Comments Sterilizing filter common rating (legal definition)除菌過濾除菌過濾-公共孔徑公共孔徑(法規定義法規定義) Prefilters have nominal ratings - each manufacturer has their own standards and methods 預過濾預過濾-額定孔徑額定孔徑,每一個制造商有自己的定義和方法每一個制造商有自己的定義和方法0.

8、22 um Durapore Surface (3,000 x)0.22 um Durapore Cross-section (600 x) “Water Loving”親水親水 Wet spontaneously (or with little effort) with water Cellulosic materials(i.e. Regenerated cellulose, mixed cellulose ester, etc.)纖維素纖維素材料材料 Polycarbonate with additives (i.e. PVPP), Modified polysulfone, Nylon

9、, Modified polyvinylidenefluoride (PVDF)聚碳化合物聚碳化合物 Applications應用應用 Filtration and sterile filtration of aqueous or aqueous / organic solutions水或水水或水/有機溶液混合的過濾和除菌過濾有機溶液混合的過濾和除菌過濾 “Water hating”疏水疏水 Not spontaneously wetwith water. Water can remain or be “tricked”to enter filter水被截流或巧妙進入膜水被截流或巧妙進入膜 P

10、TFE -Polytetrafluoroethylene, PVDF -Polyvinylidene fluoridePolypropylene, Polysulfone,Polycarbonate Applications應用應用 Solvent, acid, base, and chemical filtration溶劑溶劑,酸酸,堿和化學品過濾堿和化學品過濾 Tank/Equipment vents, process gas, fermentation inlet/exhaust filters 罐罐/設備呼吸器設備呼吸器,工藝用氣工藝用氣,發酵進氣發酵進氣/排氣過濾排氣過濾 深層過濾器

11、深層過濾器 表面過濾器特性表面過濾器特性 膜過濾器膜過濾器 Fibrous (can shed fibers)纖維纖維(有纖維脫落有纖維脫落) Difficult to give an accurate pore size rating不能給出一個確切的空徑不能給出一個確切的空徑 Thick (3 -30 mm) & often adsorptive厚度厚度(3-30毫米毫米),通常有吸附通常有吸附 Give a percentage (i.e. 30 -70%) particle reduction只能給出顆粒減少的比例只能給出顆粒減少的比例 Have the greatest capaci

12、ty較大的承污能較大的承污能力力 Examples -Microfiberglass, String-wound, pad filters例如例如-玻璃纖維玻璃纖維,線繞式線繞式,壓壓板式板式 Fibers locked together by heat or membrane coating纖維用熱粘合或膜涂布纖維用熱粘合或膜涂布 Given a nominal rating or rated by the filter it protects可以給出公稱空徑可以給出公稱空徑 Thin (1 mm or less) & Slightly Adsorptive較薄較薄(小于小于1毫米毫米),較

13、小吸附較小吸附 Give a percentage (90 -99.9%) particle reduction給出顆粒減少的比例給出顆粒減少的比例(90-99.9%) Examples -Cellulose ester coated cellulose or polyesterweb例如例如-纖維素酯纖維素酯涂纖維或聚酯片涂纖維或聚酯片 Strong, Rigid NOT brittle堅強堅強，硬不易碎，硬不易碎 Tortuous pathway曲折的通道曲折的通道 Not all the retention on top 不是都截流在膜表面不是都截流在膜表面 Very high inte

14、rnal area 非常高的內表面積非常高的內表面積 65-75% porosity 65%-75%開孔率開孔率 Size exclusion particle retention does not change with flow or pressure 大小排除顆粒截流與流速和壓力無關大小排除顆粒截流與流速和壓力無關Sterilizing filters must have 99.99999% removal 無菌過濾器必須具有大于無菌過濾器必須具有大于99.99999%截流截流 Integrity testable (diffusion &/or bubble point) 能做完整性測

15、試能做完整性測試 Produced by Casting 鑄造膜 Nylon, Cellulose (mixed ester, regenerated), PVDF, Polysulfone 尼龍,纖維素(混合酯, 改良),聚偏二氟乙烯,聚砜 Produced by Stretching 拉伸膜 PTFE 聚四氟乙烯 Mainly made by casting membrane 主要由鑄造方式制造 Can be either hydrophilic orhydrophobic 可以是親水和疏水 Rated on the size of the smallest particleit reta

16、ins 孔徑是對最小的顆粒的截留 Very thin (100 - 260 um) 非常薄（100-260微米） Adsorption depends on materials 吸附與材料相關 Examples例如 Cellulose ester纖維素酯 Regenerated cellulose改良纖維素 Nylon尼龍 Polysulfones聚砜 PVDF聚偏二氟乙烯 Slot shaped pores 狹窄的孔道 Very thin 非常薄 Often bonded to a supportmaterial 常常與支撐結合 Can have high proteinadsorption

17、 有較高的蛋白吸附 Examples例如 PTFE - Teflon聚四氟乙烯 Naturally hydrophobic天然疏水 Most hydrophobic polymer澄清(Millistak+ orPolygard)預過濾(Milligard orPolysep-II)最終(Durapore 0.22 or0.45m）深層表面膜 孔徑逐漸減小的過濾系列 Clarifying filter-澄清過濾 深層過濾 在過濾系列開始是最大的 過濾容量 Prefilter-預過濾 Prefilter-預過濾-表面過濾 中等截留，中等載量 Final filter-除菌過濾 除菌膜 最大的過濾

18、截留率Performance ofFiltration Systems過濾系統性能Non-deformable types不可變形顆粒 Resin beads or fines樹脂粒 Drug crystals藥物晶體 Carbon fines炭粒 Diatomaceous Earth (D.E.)硅燥土 Form porous permeable cakes.在膜上形成可流通的餅Deformable types可變形顆粒 Proteins蛋白 Lipids脂 Sugar complexes糖復合物 Can move through the filter, breakup and compre

19、ss into impermeablecakes.能穿過膜,破碎,壓縮* Happens with hard particles 發生在硬顆粒* Particles build up on the surface of the filter 顆粒堆積在過濾膜的表面 Happens with deformable particles 發生在可變形顆粒 Particles completely block the pore“ 顆粒完全堵塞膜“孔” Happens with little prefiltration OR particles slightly larger than the pore

20、 size 發生在缺少預過濾或顆粒略大于孔徑的情況 Happens with hard or deformable particles 發生在硬顆粒和可變形顆粒 Particles build up on the pore throat or opening 顆粒堆積在孔道或表面 Most common with biologicals 大多發生在生物化學顆粒 Gradual blockage most common 逐漸堵塞經常發生 Everything was going alright, then all of a sudden the filter blocked” 一切正常,然后突然

21、過濾器堵 塞了 流速下降 可能低于要求的流速(如灌裝速度) 壓力上升 可能超過過濾系統承受的壓差 過濾柱或管道/連接件 顆粒可能穿過過濾器 污染/堵塞下游過濾器 過濾成本改變 時間,產品和過濾器的成本 系統停止或更換過濾器 本道工序濾膜的選擇，孔徑，面積 操作條件（流速，溫度，壓力） 上道工序濾膜的選擇 產品本身的變化 產品過濾前作壓力流速實驗 使用低的起始壓差 1-2 psid or 0.1 bar 監測和調整壓差 深層過濾:10-15PSID 表面過濾和膜過濾:30PSID 不超過制造商的規格 確保過濾在合時壓力下進行 圓盤過濾器 13-293毫米直徑 傳統的濾器 可同時使用深層、表面濾膜

22、 通常用于小批量過濾 成本低 分量重 安裝困難 占地面積大Stacked Disk層疊式圓盤 Membrane bonded to both sides of a support disk 膜結合在圓盤支撐的兩邊 Support disks bonded together 圓盤支撐結合在一起 All filters work in parallel所有過濾器平行 Low hold-up volume殘液低 Easy to use & compact使用方便緊湊 Either replacement filter (Millidisk) or disposable device (Millipa

23、k) 有可替換的和一次性使用的過濾器 Non-metallic非金屬 Non-flexing structure不可變形構造 Low resistance to back pressure 承受反壓低Pleated Cartridge折疊式 Membrane held between supportingmaterial膜在支撐材料間 Thermoplastic welding熱熔 2-5 materials of construction 2-5種結構材料Good forward & reverse pressure resistance 較高的正/反壓承受能力 Large surface

24、area大過濾面積Multiply steamable多次蒸氣滅菌Variety of sizes, surface areas and o-ringconfigurations各種規格，表面積和形圈結構Can have prefilter / final filter in the samecartridge能將予過濾和除菌過濾膜結合在同一過濾芯內 濕潤O型圈，易于安裝 檢查過濾外殼方向 inlet / outlet 不要過緊，合適的套筒不需要太大的力量 避免直接用手接觸過濾器 使用前沖洗過濾器 滅菌次數： CVGL除菌濾芯： 30 SIP 30 min 135 C; 30 高壓滅菌60 m

25、in 126 C KVGL囊式除菌濾芯： 3 高壓滅菌60 min 126 C; 不能在線滅菌 Millidisk除菌濾器： 5 SIP 60 min 135 C; 5 高壓滅菌60 min 126 C 。 Aervent空氣/有機溶劑除菌濾器：150（100正向/50反向） SIP 30 min 145 C; Express聚醚砜除菌過濾器： 25 次正向（或22次正向，3次反向） 30min135 C; 一定要預先閱讀說明書-實際工藝使用次數需要確認 Preventative Maintenance 預防性維護Oxidation氧化Integrity failure完整性失敗 Filter

26、 blockage過濾膜堵塞Too many sterilization cycles太多滅菌次數Filter changeout can vary; 1 week to 1 year過濾器更換時間范圍很廣(從1周到1年)Need documentation需要文本支持Increased scrutiny應不斷進行檢查There are two major retention mechanisms in liquids在流體過濾中有兩類過濾機理 Adsorption, Size Exclusion吸附,大小排除Three types of filters are commonly used i

27、n biopharmaceuticalfiltration三類過濾器普遍使用在生物制藥 Depth, Surface, Membrane深層,表面,膜過濾Filter materials are supplied as either過濾介質以下列形式提供 Disc, Stacked Disc, Pleated Cartridge圓盤,層疊式,柱式Filters block by either過濾堵塞可以是 Gradual, Total or Cake逐漸堵塞,完全堵塞Filtration problems can be solved過濾問題可以被解決 Monitoring and optimi

28、zing are key techniques監測和優化是關鍵技術Integrity Testing Theory完整性測試理論 為什么要對除菌濾器進行完整性測試？ 完整性測試的方法有哪些？ 為什么可以用起泡點的方法代替細菌挑戰試驗？ 什么時候進行完整性檢測？ 起泡點測試原理是什么？ 擴散流測試原理是什么？ 自動完整性測試儀Common Sense通常理解 Filtration is often the most critical step in an operation 過濾通常是操作的關鍵步驟 Confirmation of manufacturers specifications 確認制

29、造規格 Detecting leaks due to o-rings, gaskets, seals 檢測O形環,墊圈,密封墊的泄漏 Assuring the correct pore size filter 確認正確的過濾孔經 Assuring integrity before sterilization 確認滅菌前完整性 Assuring integrity after steaming or autoclaving 確認蒸汽和消毒鍋滅菌后完整性Business Practice 商業慣例 Government Guidelines & Regulations法規要求 Part of co

30、rporate standard operating procedure公司標準操作規程 Auditing requirement審計需要 FDA Guideline指南(2003) Whatever filter or combination of filters is used, validation should include microbiological challenges to simulate worst-case production conditions”對于由一個或多個濾器組成的過濾系統，對它的驗證都應該包括在最差條件下進行的微生物挑戰試驗Revision of ann

31、ex 1 to EC Guide to GMP for sterile medicinal products(1997)歐盟對于無菌藥品GMP指南的附件一(1997版) “The integrity of the sterilized filter should be verified before use and should be confirmed immediately after use by an appropriate method such as a bubble point, diffusive flow or pressure hold test” 除菌過濾膜應該在使用前及

32、使用后立即采取合適的方法確認其完整性,可以采用泡點,擴散流或壓力保持的方法.FDA Guideline on Sterile Drug Products Produced by Aseptic Processing (1987)FDA對于無菌操作生產的無菌藥品的要求(1987版) “Normally, integrity testing of the filter is performed after the filter unit is assembled and sterilized prior to use.” 完整性測試通常在過濾器安裝,滅菌后使用前進行PDA Technical Re

33、port # 26PDA技術報告26版 “It generally is regarded as a cGMP requirement that filters or filter systems routinely be integrity tested both prior to and after use.” 現行的GMP要求過濾器及過濾系統在使用前及使用后均需要進行完整性測試 WHEN TO DO IT? Detect what? 什么時候做? 檢查什么？ Before sterilization faulty housings? 滅菌前 外殼的問題 Before use out-of

34、-the box failures 使用前 包裝以外的問題 After use sterilization induced failures 使用后 滅菌的影響 stress induced failures 壓力的影響分類 測試名稱 測試實施者破壞性破壞性細菌挑戰測試制造商以及客戶進行驗證時進行非破壞性非破壞性 起泡點測試，擴散測試制造商出廠時及使用者現場進行金標準Regulatory & quality organizations need datafrom both to assure reliable and predictablefilter performance法規和質量管理需要

35、數據確保可靠和預知過濾器性能 There must be a correlation between a destructive and a non-destructive integrity test 破壞性和非破壞性測試兩者必須有關聯 Regulatory requirement 法規需要 FDA Aseptic Guidelines FDA無菌指南 Validation justification for the use of a test 測試應用的驗證理由Integrity Testing Correlations完整性測試關聯What Non-Destructive Integri

36、ty Tests are Available?何種非破壞性測試合適?Bubble Point Introduction 起泡點介紹起泡點介紹Bubble point is the pressure at which gasdisplaces liquid from the largest set of filterpores and flows rapidly through the filter起泡點是氣體從濕潤的最大膜孔擠出液體快速流出時的壓力Bubble point indicates the magnitude ofthe forces holding liquid in the fi

37、lterstructure起泡點顯示過濾結構內保持液體的力的大小The oldest non-destructive integrity test最傳統的非破壞性測試泡點測試Bubble Point Test Applicability起泡點測試適用性Bubble Point Equation起泡點等式起泡點等式泡點的影響因素接觸角的影響Examples of the Effect of Contact Angle andSurfaceTension on Bubble Point Values接觸角和表面張力對起泡點值的影響實例Diffusion Testing Introduction擴散

38、流測試介紹 Gas dissolves in liquidheld in the pores of a fullywetted membrane filter.氣體溶解在完全濕潤的濾器膜孔內的液體中 A pressure differential willgive a different gasconcentration across thefilter.膜壓差造成跨膜氣體濃度差 Results in gas flowthrough the liquiddissolved in the filter pores.結果使膜孔溶解的氣體流出擴散流測試Diffusion Testing Equati

39、on擴散測試方程擴散測試方程A Reason for Diffusion Testing什么情況下采用擴散測試Diffusion Test Applicability擴散測試適用性Factors Affecting Diffusion Tests擴散測試的影響因素Example of Changed Diffusional Flow 舉例Example of Changed Diffusional Flow 舉例Example of Changed Diffusional Flow 舉例“Diffusion Testing” Summary 擴散測試總結疏水濾膜采用有機溶劑進行完整性測試的問題

40、HydroCorr Testing Hydrocorr測試測試HydroCorr Test Resistance to water intrusion水浸入法測試水浸入法測試HydroCorr Test Results測試結果HydroCorr Test Considerations測試的注意點測試的注意點 Must have a leak-free system必須在一密閉系統內進行 Filters must be dry and clean過濾芯必須干燥且干凈 It is relatively easy to Hydrocorr test “very hydrophobic” membra

41、nes such as PTFE 對于疏水性很強的濾芯,例例PTFE材質的膜材質的膜,水浸入法是相當容易水浸入法是相當容易 Successfully testing “less hydrophobic” membranes such as PVDF requires idealconditions that can be difficult to achieve in actual biopharmaceutical processes 在實際的生物制藥工藝中,測試一些疏水性不強的濾膜測試一些疏水性不強的濾膜,例例PVDF膜是比較困難膜是比較困難的的 Temperature changes s

42、hould be minimized 盡量減少溫度的變化 Reference testing is important 相關的參考比較重要 Must have a complete validation package 必須有一完整的驗證文本 Values must be comprehensively correlated to microorganism challenge 數據必須與細菌挑戰相關完整性測試失敗處理措施潤濕建議Summary總結總結 Bubble point and Diffusion (or “Forward flow”) are BOTH validintegrity

43、tests as per regulatory agencies.起泡點, 擴散流（或前向流）和HydroCorr是法規認可的完整性測試方法 The choice of an integrity test depends on the testing equipment,the filter manufacturer, the companys philosophy and the testingenvironment完整性測試的選擇倚賴測試裝置，過濾器制造商，公司體系和測試環境 Bubble point provides a direct correlation to bacterial r

44、etention, thecritical performance characteristic起泡點提供與細菌截留的直接關聯，重要的性能特性 Diffusion testing provides a sensitive way to determine integrity forlarger area filters擴散測試提供一個敏感方法來測試大面積過濾器Automated Integrity Testers自動完整性測試儀Why Do People Use Automatic Testers為何用戶會選用自動測試儀Post SIP Integrity Testing SIP后的完整性測試

45、后的完整性測試Revision of annex 1 to EC Guide to GMP for sterile medicinal products(1997)歐盟對于無菌藥品GMP指南的附件一(1997版) “The integrity of the sterilized filter should be verified before use and should be confirmed immediately after use by an appropriate method such as a bubble point, diffusive flow or pressure h

46、old test” 除菌過濾膜應該在使用前及使用后立即采取合適的方法確認其完整性,可以采用泡點,擴散流或壓力保持的方法.FDA Guideline on Sterile Drug Products Produced by Aseptic Processing(1987)FDA對于無菌操作生產的無菌藥品的要求(1987版) “Normally, integrity testing of the filter is performed after the filter unit is assembled and sterilized prior to use.” 完整性測試通常在過濾器安裝,滅菌后使用前進行PDA Technical Report # 26 PDA技術報告技術報告26版版 “It generally is regard