Abstract
The use of Cannabis sativa, or Hemp, in commercial, recreational, and pharmacological applications is on the rise in the United States and worldwide. Many of these applications have guidelines associated with them dependent on the concentration of cannabinoid molecules that keep the products classified as hemp versus marijuana or that allow the producer to comment on the purity and potency of their product. Herein, we propose a method for homogenization of hemp that results in small particle sizes, uniform samples, and does not alter the cannabinoid concentrations during processing, allowing for optimal and reproducible potency testing. Using a novel “active grinding media” we homogenized commercially available hemp to analyze approximately 100 mg samples of homogenate via sieve analysis and high-performance liquid chromatography to assess the resulting size and potency of the sample when using this methodology. When processing hemp samples with our proposed methodology, we have demonstrated the ability to produce 60.2% of all particles < 1.25 mm with increased cannabinoid recovery compared to homogenates with larger average particle sizes. Maintaining sample temperatures below 35 °C during processing, we showed that our method does not thermally induce decarboxylation reactions that would result in major cannabinoid profile changes. We have developed a method for hemp processing via homogenization that does not alter the cannabinoid profile during processing, while consistently producing small particle sizes in a uniformly processed sample. This method allows for optimal and reproducible hemp processing when evaluating hemp and hemp-based products being brought to commercial markets.
Highlights
Cannabis sativa, commonly known as “hemp”, has widestretching usages in industrial, medicinal, and agricultural applications across the world [1]
As average particle size decreased with each cycle of homogenization, we observed an increased extraction of CBD, CDBA, THC, and tetrahydrocannabinolic acid (THCA), with a significant increase in the detection of CBD and cannabidiolic acid (CBDA) (Table 2,)
In addition to providing particle sizes < 1.25 mm, the use of our active grinding media and cyclical homogenization approach did not cause decarboxylation changes to the THCA and CBDA populations seen in the hemp tested as confirmed by reverse phase HPLC (Fig. 1, Supplemental Fig. 1)
Summary
The six (1 mL samples of the 250-ppm cannabinoid) standards were added to reinforced tubes (2 mL) that were prefilled with 2.8 mm ceramic beads (Omni International, Kennesaw, GA, Cat. No 19–628). The tubes were processed on the Bead Ruptor Elite Bead Mill Homogenizer at 5 m/s for increasing durations of 10, 20, 30, 60, and 120 s. Time point, 250 μL was removed to a new 1.5 mL microcentrifuge tube and placed on ice. As a positive control, 500 μL of the 250-ppm cannabinoid standard was placed in a 1.5 mL microcentrifuge tube and heated, at 90 °C, for 180 min. All samples were filtered through a 0.2 μm spin filter and 15 μL of the filtrate was combined with 35 μL of 100% HPLC grade methanol for reverse phase HPLC analysis
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