Understanding the Complexity of Microbiology Method Validation to Meet State Specific Requirements for Cannabis.

BACKGROUND:Federal legislation still prohibits the cultivation, sale and consumption of cultivars of delta 9-tetrahydrocannabinol cannabis (>0.3%), however, as of November 2022, 39 states have legalized these products for medicinal consumption and 21 states have legalized for adult-use consumption. This state-by-state approach has produced a patch work of regulations that multi-state operators (MSO) must learn to navigate. Furthermore cannabis laboratories often lack the space and skill needed to perform method validations adding another layer of complexity. While these barriers exist, it is paramount for MSOs to demonstrate the fitness of purpose of their methods. OBJECTIVE:This review presents the complexity that a MSOs navigated in developing microbiology method validation study designs for two proprietary Real-Time PCR (RT-PCR) assays to meet four state (California, Florida, Michigan and Oklahoma) testing requirements. The testing in each state was conducted in addition to certification of the assays through the AOAC Performance Tested Method  SM (PTM) program. METHODS:Matrix studies were conducted targeting three analytes (Aspergillus spp., Salmonella spp., and Shiga toxin-producing Escherichia coli) as directed by the state regulatory authorities. For California, inclusivity and exclusivity studies were performed. The number of contamination levels, test portion replicates, and total target organisms evaluated varied by state. Culture confirmation was not performed outside of the AOAC PTM studies. RESULTS:Data from each study was collected, summarized, and provided to the state regulatory agencies. Review of the data consisted of identifying discrepant results and verification of controls. After review, each assay was certified for use in the respective state. CONCLUSION:Requirements from each of the states demonstrate the complexities MSO face and emphasize the need for a more standardized approach to streamline acceptance of alternative methods. HIGHLIGHTS:While varying state regulations can be complex, validation studies performed for the candidate methods have led to adoption across 31 states.