Verification Of Methods Following EN ISO 16140-3; Practice
Verification of methodsfollowing EN ISO16140-3; PracticeWilma JacobsMember of ISO TC34/SC9/WG3:Method validation1
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ISO 16140-3 Verification requirements for aqualitative (detection) onVerification(Food) itemverificationQualitativeEstimated LOD50 (eLOD50) eLOD50: 3 available protocols to determine the eLOD503
Implementation Verification Demonstrate competenceof the user laboratory toperform the method(obtain expected results on a(food) item used in thevalidation study) The user laboratory shall:1. review validation data forthe method,42. select 1 (food) item testedduring the validation studythat belongs within thescope of laboratoryapplication of the userlaboratory, and3. use this 1 (food) item andthe sample size used inthe validation study toperform implementationverification.
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(Food) item Verification Demonstrate the competence of theuser laboratory to perform thevalidated method with (food) itemsthat are tested in the userlaboratory. The user laboratory shall:7 select 1 challenging (food)item from each (food)category listed within thescope of validation, that isalso a (food) category that istested within the scope oflaboratory application ofthe user laboratory, and use this 1 (food) item andthe sample size used in thevalidation study to performthe (food) item verification.
Categories: Broad range of foods and other.Table A. 1: Classification of sample types& suggested target combinations for validation studiesCATEGORIESRaw milk &Heat processeddairy products milk & dairyproductsRaw meat &ready-to-cookmeat products(except poultry)Ready-to-eat,ready-to-reheatmeat productsEggs & atfishery productsFresh produce & Processed fruitsfruits& vegetablesRaw & ready-tocook fish &seafoods(unprocessed)Infant formula Chocolate,& infantbakery productscereals& confectionary8Raw poultry &ready-to-cookpoultry productsMulti-component EnvironmentalPet food &foods or mealsamples (food or animal feedcomponentsfeed production)Ready-to-eat,ready-to-reheatmeat poultryproductsDried cereals,fruits, nuts,seeds andvegetablesPrimaryproductionsamples
User laboratory: RIVM-Z&O Laboratory Detection of Salmonella according to ISO 6579-1 “a broad range of foods” plus the 3 “other” categories– 1 x Implementation verification– 8 x (Food) item verificationVerificationCategoryRaw poultry & ready-to-cookImplementation verificationpoultry products(Food) item verification(Food) item verification(Food) item verification(Food) item verification9Raw milk & dairy productsItemInoculum strain Strain sourceChicken fillet partsInfantisBroilerRaw milkHadarHumanSTMmonoPig meatEnteritidisEggAgamaShellfishVirchowHumanRaw meat & ready-to-cookMinced meat (beef/pork)meat products (except poultry)Eggs & egg productsWhole liquid egg(derivatives)Raw & ready-to-cook fish &Musselsseafoods (unprocessed)(Food) item verificationFresh produce & fruitsCantaloupe(Food) item verificationEnvironmental samples (foodor feed productionSpent irrigation water(Food) item verificationPet food & animal feedChicken feed(Food) item verificationPrimary production samples(PPS)Chicken faeces: boot socks TyphimuriumEnvironmentalSenftenbergCocoa powderChicken: boot swabs
Verification Qualitative methods Estimated LOD50 (eLOD50) determination Enumeration of the inoculum !– Plate counts, MPNs Acceptability limits10
Protocols in ISO 16140-3- Protocol 3: direct use of reference material (3-5 cfu/ml), 7 samples11
Elaborated workflow at RIVM-Z&O- Combination of Protocol 1, Protocol2, Protocol 3 (for research reasonsonly)- Aiming at a concentration forDilution A at 27 cfu/ml- Enumeration of overnight culture(ON) of inoculation strain- ON dilutions kept in fridge and usedto prepare Dilution A- Actual inoculation dilutions B, C, D,prepared from Dilution A by 1:3steps- Both plate counts and MPN (forresearch reasons only) forenumeration of 1 ml dilutions (A, B,C, D)- Testing inoculated samples (P1, P2,P3) for presence of Salmonella12
Enumeration of inoculaaantal kolonies per plaat (14 cm, 1 ml)MONSTER NRDil A is 2,3 ml -6 plus57,7 ml PFZ (verwachting27 cfu/ml)Dil A is 2,5 ml -6 plus57,5 ml PFZ (verwachting27 cfu/ml)Dil A is 2,1 ml -6 plus57,9 ml PFZ (verwachting27 cfu/ml)Dil A is 1,4 ml -6 plus58,6 ml PFZ (verwachting27 cfu/ml)13Dil. ADil. AStamafhankelijkStamafhankelijkEnt S6 op NA363781036Ent S8 op NA4731121265Ent S10 op NA3735151012Ent S11 op NA3127101353PRODUCTDil. BDil. BDil. CDil. Cis 20 ml Dil. A is 20 ml Dil. A is 20 ml Dil. B is 20 ml Dil. Bplus 40 ml PFZ plus 40 ml PFZ plus 40 ml PFZ plus 40 ml PFZMPN tablein ISO 16140-30,62 [0,15 -2,6] cfu/test portionCFU/ml Dilution D1,35 cfu/test portion
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Combined results, per ProtocolFood item: Animal feedProtocol 1Protocol 2Protocol 316Dilution AStrain S6naDilution B(1:3 of A)1/1Dilution C(1:3 of B)4/4Dilution D(1:3 of C)3/4BlankDilution AStrain S6naDilution B(1:3 of A)naDilution C(1:3 of B)3/3Dilution D(1:3 of C)4/5BlankDilution AStrain S6naDilution B(1:3 of A)naDilution C(1:3 of B)naDilution D(1:3 of C)6/7Blank0/10/10/1
Evaluation of the results, per ProtocolFood item: Animal feedInoculation strain S6(LIL)eLOD50Published validation dataor assume 1 cfu/test portionLOD50(cfu/test portion)(cfu/test portion)(cfu/test portion)(cfu/test portion)e LOD50 4 x LOD501,40,62 [0,15 - 2,6]0,70,311144AcceptedAccepted1,40,62 [0,15 - 2,6]0,560,251144AcceptedAccepted1,4 ( [3-5] )0,62 ( [3-5] )nanananaminimum 6/7minimum 6/7AcceptedAcceptedLow Inoculation Table 6/Table 8ISO 16140-3 0,5 x LIL 0,5 x LIL 0,4 x LIL 0,4 x LILnana4 x LOD50QualitativeVerificationAcceptability Limit
User laboratory: RIVM-Z&O Laboratory Detection of Salmonella according to ISO 6579-1 “a broad range of foods” plus the 3 “other” categories– 1 x Implementation verification– 8 x (Food) item verificationVerificationCategoryRaw poultry & ready-to-cookImplementation verificationpoultry products(Food) item verification(Food) item verification(Food) item verification(Food) item verification18Raw milk & dairy productsP1:ItemInoculum strain Strain sourceChicken fillet partsInfantisBroilerRaw milkHadarHumanSTMmonoPig meatEnteritidisEggAgamaShellfishVirchowHumanRaw meat & ready-to-cookMinced meat (beef/pork)meat products (except poultry)Eggs & egg productsWhole liquid egg(derivatives)Raw & ready-to-cook fish &Musselsseafoods (unprocessed)(Food) item verificationFresh produce & fruitsCantaloupe(Food) item verificationEnvironmental samples (foodor feed productionSpent irrigation water(Food) item verificationPet food & animal feedChicken feed(Food) item verificationPrimary production samples(PPS)Chicken faeces: boot socks TyphimuriumEnvironmentalSenftenbergCocoa powderChicken: boot swabs
Thank you for your attention !Any questions ?19
Verification of methods following EN ISO 16140-3; Practice Wilma Jacobs Member of ISO TC34/SC9/WG3: Method validation. Salmonella 2. ISO 16140-3 Verification r . Low Inoculation Level or assume 1 cfu/test portion Verification Results Table 6/Table 8 (LIL) eLOD. 50. LOD. 50. 4 x LOD. 50.
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(System-on-Chip). This has made verification the most critical bottleneck in the chip design flow. Roughly 70 to 80 percent of the design cycle is spent in functional verification. [1]System Verilog is a special hardware verification language to be used in function verification. It provides the high-level data structures available
new approaches for verification and validation. 1.1. Role of Verification and Validation Verification tests are aimed at "'building the system right," and validation tests are aimed at "building the right system." Thus, verification examines issues such as ensuring that the knowledge in the system is rep-
Design vs. Verification Verification may take up to 70% of total development time of modern systems ! This ratio is ever increasing Some industrial sources show 1:3 head-count ratio between design and verification engineers Verification plays a key role to reduce design time and increase productivity 10 IC Design Flow and Verification
7.2 Selection, verification and validation of methods 7.2.1 Selection and verification of methods 7.2.2 Validation of methods 7 7 Process requirements . . individual test e.g. garlic, hops, tea, spices, cranberry 24. Representative samples Pesticide residues Commodity group 1. High water content
System verification refers to checking that a delivered system meets its requirements, usually by testing (although other verification methods are employed where testing is costly or impossible). Note that the validation activity (as defined here) must precede verification, as pointed out in O'Grady.2 Unfortunately,
complements the IEEE 2800 test and verification framework with specifications for the equipment, conditions, tests, modeling methods, and other verification procedures - may specify design and as -built evaluations procedures for verification of plant-level capabilities and performance - may also specify verification procedures for IBR .
HIGH RISK BAKING Although most cakes and biscuits are classed as low risk products, some fillings and finishes are more high risk. Fresh cream, some cheese cakes and royal icing made from raw egg whites are all high risk and require extra thought to ensure they are prepared safely. Cakes that require refrigeration must be kept at or below 8 C at all times with limited time out of temperature .
