AOAC CASP Cannabinoids ERP October Method Book

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(12) CBL. Cannabicyclol

(13) CBC. Cannabichromene

(14) CBCA. Cannabichromenic Acid

(g) Analytical Standards – Certified Reference Materials; 2.5 mg; Toronto Research Chemicals

(1) CBE . Cannabielsoin

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PREPARATION OF TEST SAMPLES & STANDARD SOLUTIONS

Protect solutions from direct sun or UV light. All sample preparation procedures, when possible, should 125 be done in a fume hood while wearing gloves and eye protection. 126 (a) Preparation of calibration curve. – Prepare a calibration curve inmethanol from certified analytical standards for each cannabinoid of interest. Calibration curves may be prepared individually, or mixed. The linear range of each compound’s prepared calibration curve should range from 0.1% w/v to 0.00001% w/v. Points at either extremity of the curve which reduce the curve’s linearity (R 2 < 0.99), or whose signal-to-noise ratio is less than 10 should not be used. All calibration curve solutions should be prepared using a gas-tight syringe using methanol as a diluent. Calibration curves are plotted as Peak Area vs. Concentration (% w/v), with the x-intercept set to 0.0 and the

127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144

line of best fit following the equation:

Peak area = (Slope)(Concentration)

(b) Preparation of test samples .

(1) For solid chocolate, including chocolate bars and chocolate chips – Homogenize the solid chocolate sample using a mortar and pestle. Accurately weigh 0.08000 g of chocolate

(with a tolerance of +5%) into a glass test tube and record the mass.

(2) For chocolate truffles – Place the entire truffle (either whole or cut into pieces, if necessary) into a closed container, such as a 50 mL centrifuge tube. Place in a 65°C bead bath until the chocolate is melted, about 10 minutes. Using a lab spatula or stir bar, thoroughly mix the melted chocolate. Accurately weigh 0.08000 g of chocolate (with a

tolerance of +5%) into a glass test tube and record the mass.

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DETERMINATIONS

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(a) Preparation of test sample for HPLC analysis. – Heat the weighed sample using a 65°C bead bath until the sample has fully melted, about 5-7 minutes. Before the sample cools, immediately add 10 mL of the Extraction Solution to the test tube. Vortex the sample for 30 seconds. Sonicate the

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