AOAC CASP Cannabinoids ERP October Method Book

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(1) Certified Reference Materials – A spiking solution containing equal amounts of the following CRMs from Cerilliant Corp: Δ 9-THC, Δ 9-THCA, CBD, CBDA, Δ8 -THC, each with a certified concentration of 1.000 mg/mL, and different lot numbers than used for (2) In-House ReferenceMaterials – Four in-house reference materials were created by mixing the four negative control materials with controlled amounts of cannabis resin. The cannabis resin was tested for potency using a validated method. As such, concentrations in the in-house reference materials can be calculated based on a mass-balance approach. All test materials were weighed into 50 mL centrifuge tubes. Mixing occurred in a 50°C water bath to prevent decarboxylation of the acidic cannabinoids with constant stirring calibration curves.

with a spatula for 15 minutes. The details of the mixing are in Table 1.

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SCOPE

The method is suitable for the quantification of 0.005 – 6.25 % w/w cannabinoids in dark, milk, and white 71 chocolate. For some cannabinoids, this range may be extended even further based on instrument 72 linearity . This method validation study directly analyzes Δ9 - THC, CBD, Δ9 - THCA, CBDA, CBN, and Δ8 -THC, 73 and can precisely monitor an additional 9 impurity cannabinoids, including CBE, the major oxidative 74 compound of CBD (Figure 1). This method has been validated to include chocolate bars, chocolate chips, 75 and chocolate truffles, but does not include chocolates with nuts, fruit, or other ingredients which will not 76 fully homogenize upon melting. This method is suitable for HPLC-DAD as well as HPLC-HDR-DAD. The 77 instrument’s minimum linear range must be equal to 0.6 – 400 ppm to accurately capture cannabinoid 78 concentrations of 0.008 – 5% w/w in chocolate samples. The validation data captured here was collected 79 using an HDR-DAD detector. 80

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PRINCIPLE

Chocolate samples are melted and extracted with 90:10 methanol:chloroform. After a lipid-removal step, 82 filtered samples are analyzed by C18 gradient reverse-phase high-pressure liquid chromatography with a 83 diode array detector. 84

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APPARATUS & MATERIALS

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(a) High-pressure liquid chromatograph (HPLC) – Liquid chromatograph with automated sampler, quaternary gradient pump, de-gas module, column heater, and diode array detector (DAD) or high

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