无菌系统验证培训

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1、TechnicalAseptic Line Validation Training无菌系统验证培训无菌系统验证培训October 27, 2005October 27, 2005Steven SunSteven SunTechnicalSupply Chain Management .2ContentPurposeScopeDefinitions Equipment Performance ValidationLine Qualification Installation / Operation qualificationProcess Validation Operational Valid

2、ation PreparationTechnicalSupply Chain Management .3Purpose qvalidation of a new aseptic line installation,qvalidation of existing aseptic lines at new contract packers,qvalidation of existing aseptic lines at existing contract packers that didnt perform the validation according to TCCCqvalidation o

3、f a new design or size of bottle or closure other than those used in the original or previous line validation,TechnicalSupply Chain Management .4Purpose qvalidation of a more microbiologically sensitive high acid beverage than the one for which the line has been validated,qyearly revalidation of an

4、existing aseptic line,qrevalidation of an aseptic line following any change that may impact the aseptic processTechnicalSupply Chain Management .5ScopeqThis document applies to all aseptic lines, which are filling high acid (pH 4.4) and low acid beverages for The Coca-Cola Company in PET or other ty

5、pes of plastic (e.g. HDPE). TechnicalSupply Chain Management .6Definitions qBioburden qCommercial SterilityqClass 100 air qualityqHEPA, ULPAqQualificationqValidationqVerificationTechnicalSupply Chain Management .7DefinitionsqLow Acid product pH4,4qHigh Acid product pH =4.4qLog reduction = Log (Initi

6、al Count/Surviving count). qOne surviving count out of a million is log 6 reduction.qIn general Low acid product log reduction is 1 unit .igher than High acid product.qUnder Commercial Sterility the consumer acceptance rate is minimum 1 spoiled product in 10,000. Not to be taken as production standa

7、rd.qValidation involves various challenging microbiological loading tests to measure up the log reduction and production simulation tests with culture media and product to verify sterility.TechnicalSupply Chain Management .8Definition qCIP SIP qCOP/SOP.qSterile zoneqMicrobiological Failure rate qSys

8、tem Failure rateTechnicalSupply Chain Management .9Criteria qCIP cleaning requirementsqSterile zone cleaning requirements (COP)qSterile zone sterilization requirements (SOP)qSIP requirementsqPackage decontamination requirementsqBottle/closure rinse waterqWater used for product change overqPressure d

9、ifferential monitor and controlqHEPA/ULPA filter installationsTechnicalSupply Chain Management .10PreparationqLine EfficiencyqOperation qUtilityqMaterialqIndicate organisms TechnicalSupply Chain Management .11Material PreparationIndicator organismIndicator organismGrowth medium Growth medium Incubat

10、ion temperature Incubation temperature timetimeScopeScopeBacillus Bacillus subtilissubtilisPlate Count Agar (e.g. Plate Count Agar (e.g. OxoidOxoid CM 325)/ TSA CM 325)/ TSA 35C 5 days35C 5 daysS2, S3, S4S2, S3, S4Bacillus Bacillus stearothermophilusstearothermophilusPlate Count Agar (e.g. Plate Cou

11、nt Agar (e.g. OxoidOxoid CM 325)/ TSA CM 325)/ TSA35C 5 days35C 5 daysTotal count (TCCC)Total count (TCCC)TGE /NATGE /NA35C 5 days35C 5 daysS5, S8, S10, S5, S8, S10, S11S11Yeast and mold Yeast and mold (TCCC)(TCCC)M-GYM (membrane filtration method)M-GYM (membrane filtration method)OSA (pour plate me

12、thod)OSA (pour plate method)28C 5 days28C 5 daysS5, S8, S10, S5, S8, S10, S11S11TechnicalSupply Chain Management .12Indicate organisms PreparationqStorageqDilutionqConcentration confirmationqGrads dilutionqInoculationqTransferqTestqMicrobe testTechnicalSupply Chain Management .13ValidationqS1 Check

13、claimed sterilizing effect on bottles, closures/alu foil at supplier qS2 Strip Test - decontamination of clean room/sterile zoneqS3 Bottle/Closure/Alu-foil decontamination external surfaces qS4 Bottle/Closure/Alu-foil decontamination internal surfaces qS5 Environment and external surfaces/sterile zo

14、ne TechnicalSupply Chain Management .14ValidationqS6 Check asepsis level - pasteurizer, aseptic tank, filling, capping qS7 Check asepsis level from pasteurizer, filling, capping qS8 Check asepsis level from pasteurizer, filling, capping + Asepsis of environment and process equipment surfaces during

15、routine operationsqS9 Transportation test package integrityqS10 Validation of line capability for commercial production of different types of beverage +Validation of performance of clean room/sterile zone after breaking asepsis +Control of residual content of sterilant in bottles + control of cappin

16、g stabilityqS11 Validation of line capability for commercial production of different types of beverage during long production runsTechnicalSupply Chain Management .15S1 pre-shipment test on sterilizing effect on bottle and capqThis test can be conducted using the actual line OR a laboratory set up w

17、ith all decontamination parameters set to operate identical to the real aseptic line using all reference bacteria. See next slide for a typical lab setup.qBrief description inoculate 100 bottles and 100 caps with specific quantities of reference bacteria. Go through the decontamination process and m

18、easure up the surviving bacteria to obtain the log reduction and compare with specification requirement.qNo shipment allowed until passing the test.TechnicalSupply Chain Management .16S1 Validation laboratory set upTechnicalSupply Chain Management .17q30 Strips preloaded with designated counts of Ba

19、cillus Subtilis were attached on different marked locations of the sterile zone of the aseptic filling chamber and conduct a COP/SOP in straight accordance to the designed specification.qExtract and culture from each strip all the surviving microbes and to obtain the log reduction and compare with s

20、pecification requirement. ALL STRIPS MUST PASS.S2 strip test to measure COP/SOP effectivenessTechnicalSupply Chain Management .18FC2FC4FB8FB7FM12FM11FM13FM15FM17FM16FM21AT5BR1BR2BR4BR6-1BR8BR9BR11BR13BS2AT1AT3BR12AS4AS6FC1CR11CR8FC6S2 Sampling Location TechnicalSupply Chain Management .19S3 External

21、 Package (bottle and cap) decontaminationqIndicator organism Bacillus subtilis.qMark and Inoculate 120 bottles 30 on bottom, 30 on body, 30 on neck finish and 30 under the neck and 30 caps on the top.qConduct usual CIP,SIP/COP and SOP in straight accordance to the designed specification when line is

22、 running smooth at its normal speed pass the inoculated bottle and cap through the system.qExtract and culture from each bottle and cap by swabbing and membrane culture to determine any surviving microbes and to obtain the log reduction and compare with specification requirements.TechnicalSupply Cha

23、in Management .20S4 Internal Package (Bottle and Cap) decontaminationqIndicator organism Bacillus subtilis.qInoculate by pipetting the interior of two sets of 100 bottles and 100 caps at different level of initial count.qGet ready uncontaminated same quantity of uncontaminated bottle and caps. qCond

24、uct usual CIP,SIP/COP and SOP in straight accordance to the designed specification when line is running smooth at its normal speed pass the inoculated bottle/uninoculated cap and uninoculated bottle/inoculated cap through the system and fill with sterile water.qPerform membrane filtration to determi

25、ne any surviving counts in each filled bottle sample and to obtain the log reduction for comparison with standard.TechnicalSupply Chain Management .21Challenge test and performance testqTest S1 to S4 all requires inoculation of an indicator microbe are designated as challenged test to verify the ste

26、rilizing effect of the system.q From S5 onward no inoculation is required, the test condition is same like production condition these tests measure consistency of line performance to deliver commercial sterility.qIt is of extreme importance that the line is set to run at all its design parameters du

27、ring the validation and after passing the validation when moving on to commercial production to keep the same settingsTechnicalSupply Chain Management .22S5 Environment TestqPerform this test after CIP/SIP/COP/SOP to evaluate the asepsis of the environment and processing equipment equipments externa

28、l surface.qTake FIVE 1 cubic metre sample of air from the sterile zone and perform yeast and Mold count.qSwab test various equipment external surface including filling valve, capping head, rinser jets after bottle decontamination etc and analyze for yeast and mold.qMeasure sterile zone particle coun

29、t.qSmoke test to confirm laminar air flow.qCompare result with specifications.TechnicalSupply Chain Management .23S6, S7, S8 first, second and third media fillS6 (first media fill)qS6 is to validate the lines total asepsis level from pasteurization to aseptic tank to filling and capping.qMedia used

30、Linden Grain at the designated pH through the product sterilizer and kept in aseptic tank.qSame procedure as normal production startup and with the line running stable at its normal speed - fill 4500 bottles with Linden Grain after 24 hrs, 48 hrs and 72 hrs product storage in the Aseptic tank.qIncub

31、ate all bottles at 25 to 30 deg C for 14 days, invert the bottles after 3 days to ensure all internal surface will contact the media. After incubation analyze 135 bottles for TAB. Use 2 pallets of S6 test product for S9 transportation test.qContaminated bottle will either be swollen, becoming cloudi

32、ng or turbid and/or with gas formation and mold like flocculation. ALL bottles MUST PASS.TechnicalSupply Chain Management .24S7 (Second media fill)qS7 is to validate the lines total asepsis level from pasteurization to filling and capping.qMedia used Same Linden Grain at the designated pH.qFill same

33、 condition as normal production 13,500 bottles with Linden Grain record any stoppages.qSame incubation and observation as S6.qUse 2 pallets of S7 test product for transportation test (S9).TechnicalSupply Chain Management .25S8 (third media fill)qS8 is an extension of S7 with simulated routine operat

34、ing conditions.qSame as S7 fill 6,500 bottles with Linden Grain.qStop the line for 60 minutes. Without doing any cleaning and sanitation, break the asepsis of the sterile zone.qResume filling an and fill 7,000 bottles. Use 2 pallets of S8 test product for S9.qDo Air quality and internal surface Swab

35、 test same procedure as S5.qSame incubation and observation as S6.TechnicalSupply Chain Management .26S9 (Transportation test)qThis test is to investigate the effect of storage and handling condition on package integrity during transportation of microbiological sensitive products in a distribution t

36、ruck.qTwo trucks each carrying 3 pallets of Linden Grain test product from Tests S6, S7, S8 under normal local transportation condition with a minimum distance 200 km plus 80 km on a hilly road.qSame Loading and Unloading condition as usual.qRecord daily highest and lowest temperatures.qDo same visu

37、al inspection after 7 days incubation.qNot any packaging faults.qTest 10 samples from each layer for opening torque.TechnicalSupply Chain Management .27Validation acceptance criteria S1 to S4Challenge testTechnicalSupply Chain Management .28Validation acceptance criteria S5 to S11Operational and Production testTechnicalSupply Chain Management .29TechnicalSupply Chain Management .30