Journal:A proposed method of sample preparation and homogenization of hemp for the molecular analysis of cannabinoids

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Full article title A proposed method of sample preparation and homogenization of hemp for the molecular analysis of cannabinoids
Journal SN Applied Sciences
Author(s) Morehouse, Zachary P.; Ryan, Gabriella, L.; Proctor, Caleb M.; Okparanta, Akelachi; Todd, Will; Bunting, Derrick; White, Taylor; Parez, Steven; Miller, Blessida; Colon, Veronica; Easparro, Brandon; Atwood, James; Park, YoungChul; Nash, Rodney J.
Author affiliation(s) Omni International, Jeevan Biosciences, Georgia State University, AMERICANNA Laboratories
Primary contact Email: Online form
Year published 2021
Volume and issue 3
Article # 762
DOI 10.1007/s42452-021-04747-6
ISSN 2523-3971
Distribution license Creative Commons Attribution 4.0 International
Website https://link.springer.com/article/10.1007%2Fs42452-021-04747-6
Download https://link.springer.com/content/pdf/10.1007/s42452-021-04747-6.pdf (PDF)
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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 (HPLC) 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.

Keywords: hemp, hemp processing, hemp grinding, homogenization, sample preparation, cannabinoid detection, THC, CBD

Background

Cannabis sativa, commonly known as “hemp,” has broad usages in industrial, medicinal, and agricultural applications across the world.[1] Currently, the applications of both hemp and marijuana are being explored in greater detail than previously seen by pharmaceutical and therapeutic applications.[2][3][4] The naturally occurring cannabinoid molecules found in hemp are the basis for its pharmacologic activity and viewed as a presumptive area of increasing importance in the field of pharmacology.[2][3] In addition to the medical application of cannabinoid-based compounds, the use of cannabinoid-infused products is on the rise for recreational usage.[5]

As states across the United States begin to legalize the sale and usage of marijuana and cannabinoid-derived products, there is an increased longing for products that will induce the reported psychoactive or therapeutic effects of these cannabinoids.[2][5][6] With demand for these products exponentially rising, the hemp industry has grown to a billion-dollar industry crossing recreational and medicinal markets for cannabinoids and cannabinoid-derived products.[5] However, as the industry has dramatically expanded in recent years, regulations and best-practice methods have not been keeping up.[6] Currently, there are a variety of unregulated techniques used in hemp processing for sample preparation and the assessment of products for cannabinoid concentrations.

Testing for the cannabinoids Δ9-tetrahydrocannabinolic acid (THCA) and cannabidiolic acid (CBDA), and their decarboxylated counterparts Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD), are critical in evaluating hemp-based products being brought to market.[7] In the U.S., hemp-based products must remain under certain concentrations of THCA and THC to remain labeled as hemp versus marijuana, a key determination in many areas given the legality of hemp versus marijuana in many municipalities.[8] Additionally, the CBDA and CBD concentrations within products have been related to their analgesic potency, thus accurate measurement and reporting of these values are vital to proper labeling and pricing of these products to protect both the supplier and consumer.[6][7][8]

Herein, we report a method for sample preparation of hemp involving bead-mill-based homogenization using a novel “active grinding media” that provides uniform processing of hemp samples with reproducible particle sizes and no alterations to the carboxylation states of THCA or CBDA. This method allows for accurate potency testing of hemp and hemp-based products through a homogenization that results in a completely processed, homogenous sample.

Methods

Sample preparation and homogenization

References

  1. ↑ Russo, Ethan B. (21 August 2007). "History of Cannabis and Its Preparations in Saga, Science, and Sobriquet" (in en). Chemistry & Biodiversity 4 (8): 1614–1648. doi:10.1002/cbdv.200790144. https://onlinelibrary.wiley.com/doi/10.1002/cbdv.200790144. 
  2. ↑ 2.0 2.1 2.2 Devsi, Alykhan; Kiyota, Brett; Ouellette, Theophile; Hegle, Andrew P.; Rivera-Acevedo, Ricardo E.; Wong, Jasper; Dong, Ying; Pugsley, Michael K. et al. (1 December 2020). "A pharmacological characterization of Cannabis sativa chemovar extracts" (in en). Journal of Cannabis Research 2 (1): 17. doi:10.1186/s42238-020-00026-0. ISSN 2522-5782. PMC PMC7819338. PMID 33526117. https://jcannabisresearch.biomedcentral.com/articles/10.1186/s42238-020-00026-0. 
  3. ↑ 3.0 3.1 Amin, Md Ruhul; Ali, Declan W. (2019), Bukiya, Anna N., ed., "Pharmacology of Medical Cannabis" (in en), Recent Advances in Cannabinoid Physiology and Pathology (Cham: Springer International Publishing) 1162: 151–165, doi:10.1007/978-3-030-21737-2_8, ISBN 978-3-030-21736-5, http://link.springer.com/10.1007/978-3-030-21737-2_8 
  4. ↑ Ebbert, Jon O.; Scharf, Eugene L.; Hurt, Ryan T. (1 December 2018). "Medical Cannabis" (in en). Mayo Clinic Proceedings 93 (12): 1842–1847. doi:10.1016/j.mayocp.2018.09.005. https://linkinghub.elsevier.com/retrieve/pii/S0025619618307092. 
  5. ↑ 5.0 5.1 5.2 Kumar, Navin; Puljević, Cheneal; Ferris, Jason; Winstock, Adam; Barratt, Monica J. (1 December 2019). "Cannabis use patterns at the dawn of US cannabis reform" (in en). Journal of Cannabis Research 1 (1): 5. doi:10.1186/s42238-019-0003-z. ISSN 2522-5782. PMC PMC7815050. PMID 33526080. https://jcannabisresearch.biomedcentral.com/articles/10.1186/s42238-019-0003-z. 
  6. ↑ 6.0 6.1 6.2 Abuhasira, Ran; Shbiro, Liat; Landschaft, Yuval (1 March 2018). "Medical use of cannabis and cannabinoids containing products – Regulations in Europe and North America" (in en). European Journal of Internal Medicine 49: 2–6. doi:10.1016/j.ejim.2018.01.001. https://linkinghub.elsevier.com/retrieve/pii/S0953620518300013. 
  7. ↑ 7.0 7.1 Coogan, Thomas A. (1 December 2019). "Analysis of the cannabinoid content of strains available in the New Jersey Medicinal Marijuana Program" (in en). Journal of Cannabis Research 1 (1): 11. doi:10.1186/s42238-019-0011-z. ISSN 2522-5782. PMC PMC7819311. PMID 33526081. https://jcannabisresearch.biomedcentral.com/articles/10.1186/s42238-019-0011-z. 
  8. ↑ 8.0 8.1 Mead, Alice (1 May 2017). "The legal status of cannabis (marijuana) and cannabidiol (CBD) under U.S. law" (in en). Epilepsy & Behavior 70: 288–291. doi:10.1016/j.yebeh.2016.11.021. https://linkinghub.elsevier.com/retrieve/pii/S1525505016305856. 

Notes

This presentation is faithful to the original, with only a few minor changes to presentation. Some grammar and punctuation was cleaned up to improve readability. In some cases important information was missing from the references, and that information was added. Nothing else was changed in accordance with the NoDerivatives portion of the license.

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