AOAC ERP MICRO AUGUST 2018

OMAMAN-44 B: Collaborative Study Protocol Expert Review Panel Use Only August 2018

1.0 1 2 Peel Plate EB (Enterobacteriaceae) is a medium used for detection and enumeration of 3 Enterobacteriaceae bacteria in food, serial dilutions of food, and environmental sponge samples. The 4 test is based on bile salt selective agar, glucose, and multiple colorimetric enzyme substrates to support 5 growth and colorimetrically identify the growth of the family of Enterobacteriaceae bacteria. The media 6 also contains gelling and wicking agents which absorb and diffuse the sample. After sample or sample 7 dilution is added, plates are incubated for up to 24 to 48 h at 37 °C ± 1°C. Peel Plate EB tests are 8 intended for microbiological laboratories, but may also be used by food quality stakeholders such as 9 farmers, milk processors, and water municipalities. The method sensitivity is greater than 1 colony 10 forming units per milliliter (>1 CFU/mL) of test sample. The accurate quantitative range is defined as 1 to 11 150 CFU/mL. 14 15 The Charm Peel Plate EB will be validated using a harmonized approach per Appendix J: AOAC 16 INTERNATIONAL Methods Committee Guidelines for Validation of Microbiological Methods for Food and 17 Environmental Surfaces (Appendix J; 1) for AOAC Official Methods SM status and ISO/FDIS 16140-2:2016: 18 Microbiology of the food chain — Method validation — Part 2: Protocol for the validation of alternative 19 (proprietary) methods against a reference method ( ISO 16140; 2) for MicroVal certification. Q 20 Laboratories, Inc. (Cincinnati, OH) will conduct the study as the MicroVal Expert Laboratory. This 21 protocol describes the AOAC validation requirements only. 22 23 The Official Methods of Analysis (OMA) program requires a Single Laboratory Validation (SLV) study, an 24 Independent Laboratory Study and a Collaborative Study (CS). For this validation, Q Laboratories 25 satisfies the AOAC independent laboratory requirement. The SLV includes matrix studies for each 26 claimed food group and inclusivity/exclusivity testing. The manufacturer (Charm) will supplement the 27 other matrices in those food groups to claim each matrix. The CS includes a matrix study on one of the 28 claimed matrices. A minimum of 8 laboratories reporting valid data is required. The reference methods 29 for the study are as follows: 1) food matrixes : ISO 21528-1:2017 – Microbiology of the food chain -- 30 Horizontal method for the detection and enumeration of Enterobacteriaceae – Part 1: Detection of 31 Enterobacteriaceae (3) and ISO 21528-2:2017 – Microbiology of the food chain -- Horizontal method for 32 the detection and enumeration of Enterobacteriaceae – Part 2: Colony count technique (4); 2) chicken 33 carcass rinse : U.S. Department of Agriculture Food Safety and Inspection Service Microbiology 34 Laboratory Guidebook 3.02 (MLG 3.02; 5) with ISO 21528:1 & 2; and 3) stainless steel : ISO 18593:2004, 35 Microbiology of food and animal feeding stuffs -- Horizontal methods for sampling techniques from 36 surfaces using contact plates and swabs (6) with ISO 21528:1 & 2. ERP Use Only Peel Plate EB – Method 12 13 2.0 Validation Overview for the Peel Plate EB

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Analytes:

Enterobacteriaceae

Heat Processed Milk and Dairy Products:

Pasteurized whole milk, butter, non-fat dry

milk, vanilla ice cream