Jack Rowe Certified Laboratories, Inc.

Bridging Science with Service Since 1926

 AOAC Performance Tested Method (RI)  AOAC Official Methods of Analysis

‣ ‣ ‣ Rapid increase in Proprietary Methods ‣ Speed to market critical AOAC OMA ‣ ‣ ‣ ‣ Higher Cost, More Time Performance Tested Methods Kicked off in 1991 Opportunity to achieve some level of validation in less time Often serves as a “pre-collaborative” until full OMA is completed

   Matrix Study

One laboratory 1-4 Foods or Surface Inclusivity, Exclusivity, Robustness, Stability and Lot to Lot Variation

 Pre-Collaborative Study and Collaborative Study  Harmonization Collaborative Study

     Selection of matrices associated with pathogen Natural vs. Artificial contamination Strain types Competing flora Fractional results

 Quantitative • • • 8 laboratories minimum, 1-6 matrices 4 inoculation levels/matrix 2 test portions per level per matrix per method  • • • • Qualitative 10 laboratories minimum, 1-6 matrices, 3 inoculation levels per matrix, 6 test portions per level per matrix per method Method compared to Reference Method

  

FDA BAM USDA FSIS AOAC

Official Methods of Analysis 2011.03

Salmonella in a Variety of Food

Validated Matrices Foods/Eggs and Egg Products, Eggs and Egg Products/Liquid Eggs, Eggs and Egg Products/Eggs, Dairy Products/Ice Cream, Vegetables/Spinach, Spinach/Fresh Spinach, Fish/Shrimp, Peanuts/Peanut Butter, Meat and Meat Products/Pork, Dairy Products/Milk, Milk/Whole Milk, Foods/Fish, Nuts and Nut Products/Pecans, Baked Goods/Cake Mix, Grains/Dry Pasta, Milk/Dried Milk, Fruit Juices/Orange Juice, Pet Foods, Water : Nuts and Nut Products/Peanut Butter, Meat and Meat Products/Turkey, Meat and Meat Products/Roast Beef, Meat and Meat Products/Pork Sausage, Meat and Meat Products/Chicken, Spices and Condiments/Black Pepper, Eggs and Egg Products/Dried Egg Yolks, Cacao Bean and Its Products/Chocolate, Foods/Fruits and Fruit Products,

Domestic Imports U.S. food import volume, by food group Food group 1999 Volume of U.S. food imports 1/ Live meat animals (1000) 2/ Meats Fish and shellfish Dairy Vegetables Fruits 3/ Nuts Coffee and tea Cereals and bakery Vegetable oils Sugar and candy Cocoa and chocolate Other edible products Beverages (1000 KL) 4/ 6,058.4

1,439.0

1,677.8

242.5

4,767.5

8,193.8

272.0

1,709.4

6,687.6

2,320.8

322.1

923.7

705.9

2,854.9

2012 7,908.0

1,335.8

2,354.2

209.8

8,545.0

11,569.6

443.0

2,093.8

10,184.4

5,178.1

3,730.6

1,237.4

1,447.3

5,820.1

 FDA will accept all matrices validated by the manufacturer for the AOAC study  Other matrices must be verified by laboratory to determine if the method is suitable for the matrix

       Amaranth Coconut Cumin Egg Cracklet Hing Powder Tahina/Sesame Seeds Octopus

Spike level must not exceed 30 cells (25 gram sample) Seven successful validations must occur (No false negatives or positives)

 Salmonella culture Atypical or Typical  Serial dilution in Phosphate Buffer  Plated in Duplicate 1ml and 0.1ml

 Selective Agar Hektoen or XLD

 Count plates after incubation per BAM Example Dilution 4 125 Dilution 5 14 Dilution 4 113 Dilution 5 12 125 + 113 / 2 = 119 cfu/ml

 Dilution 4 tube contains 119 cfu/ml  X ml x 119 cfu/ml = 30 cfu  30 cfu/119 cfu/ml = X ml  X = 0.252 or 250 ul

Prepare sample per BAM (or BioMerieux) instructions Weigh out 25 gram sample and spike with calculated volume Plate the calculated spike volume in duplicate on selective agar

 Incubate per normal protocol    Confirm counts from spike volume do not exceed 30 cells Control sample must produce a negative result. If the sample is “presumptive positive” it contains an unspecified amount of Salmonella and demonstration of < 30 cfu/ 25 gram sample is not possible Abandon Verification

    All controls must give appropriate reactions Positive Cultural Control Negative Cultural Control Diluent Blank  The Presumptive Positive spike must be confirmed following reference method

        Biochemical Identification of Spike, Negative and Positive Control Media preparation logs for all media used Autoclave Sterilization Records for all media Media Sterility records for all media used Balance Calibration Check Records Temperature Records for all Appliances Water Quality Records Confirmation serves as Document of Productivity