AOAC CASP Cannabinoids ERP October Method Book

Quantitating Cannabinoids in Edible Chocolates

In support of SMPR2017_019 Quantitating Cannabinoids in Edible Chocolates Liquid Chromatographic Separation with Photodiode Array Detection (Applicable for the quantitation of 17 phytocannabinoids in edible chocolates [white, dark and milk] from 0.008 – 4 % w/w) James W. Favell 1,2 , Ryan Hayward 1 , Seamus Riordan-Short 1 , Nahanni Sagar 1 , Rob O’Brien 1,2 , and Matthew Noestheden 1,2 * 1 Supra Research and Development, 106-2293 Leckie Rd. Kelowna, BC, V1X 6Y5 2 Department of Chemistry, I.K. Barber School of Arts and Sciences, University of British Columbia, Charles E. Fipke Centre for Innovative Research, 3247 University Way, Kelowna, BC V1V 1V7 * Corresponding author See Tables 1-3 for the results of the single laboratory validation in milk, white and dark chocolate that support the acceptance of the proposed method with respect to SMPR2017_019. A. Principle Phytocannabinoids are extracted from white, milk or dark chocolates by adding methanol and heating the resulting solution to 50 °C with ultrasonic-assisted extraction. Extracts are cold stabilized for two hours prior to analysis. Analyte separation is achieved by liquid chromatography, followed by detection using a photodiode array. Other detectors are viable alternatives, provided they meet the standards set out in SMPR2017_019. B. Apparatus, Reagents & Materials Equivalent apparatus, reagents and materials may be substituted. Apparatus a) Micropipettes — adjustable volume, with maximum capacities of 20 μL, 200 μL, 1000 μL and 5000 μL b) Analytical balance — should be capable of achieving 0.2 mg linearity, 0.1 mg readability and 0.1 mg repeatability c) Top loading balance — should be capable of achieving 0.02 g linearity, 0.01 g readability and 0.01 g repeatability d) Centrifuge — must be able to reach 3,000 rcf with 15 mL centrifuge tubes e) uHPLC system — the pumping system should possess a pressure limit of at least 1,000 bar with a flow accuracy of at least 0.1%, a thermostated sample compartment and column oven and a detector capable of meeting the method performance criteria outlined herein. f) HPLC column — must be able to resolve 17 phytocannabinoids and ibuprofen in the specified matrices (resolution could be strictly via chromatography, or in tandem with the detector [e.g., with a mass spectrometer, where chromatographic resolution of all target analytes is not required if they are mass resolved]). g) Software — should have data acquisition and processing capabilities, including auto and manual integration functions h) Heated ultrasonic bath — must reach a stable temperature of 50 ± 5 °C

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