Extractions of Medical Cannabis Cultivars and the Role of Decarboxylation in Optimal Receptor Responses, Melissa M. Lewis-Bakker et al., 2019

Extractions of Medical Cannabis Cultivars and the Role of Decarboxylation in Optimal Receptor Responses

Melissa M. Lewis-Bakker, Yi Yang, Rupali Vyawahare, and Lakshmi P. Kotra

Cannabis and Cannabinoid Research, 2019, Volume X, Number X, 2019
Mary Ann Liebert, Inc.
DOI: 10.1089/can.2018.0067



Introduction : Phytocannabinoids, characteristic compounds produced by medical cannabis, interact with cannabinoid (CB) receptors (CB1 and CB2) as well as other receptor systems to exhibit their corresponding pharmacological effects. In their natural form, CBs such as D9-tetrahydrocannabinolic acid and cannabidiolic acid are inactive at these receptors, while their decarboxylated forms (D9 tetrahydrocannabinol and cannabidiol, respectively) are potent ligands at CB receptors. Thus, extraction and processing of medical cannabis for active constituents are important.

Purpose and Methods : Patients consuming medical cannabis often have limited alternative treatment options and in recent years, medical cannabis extracts have been popular as a substitute for dried cannabis plants, despite limited studies on these derivatives. We investigated three disparate cannabis cultivars and compared four chemical extraction methods head to head, viz. Soxhlet, ultrasound-assisted supercritical fluid, and microwaveassisted extractions, for their efficiency. We further characterized the chemical compositions of these extracts.

Results : Microwave extraction consistently produced completely decarboxylated phytocannabinoid extracts. Factors such as temperature and exposure time play important roles in the decarboxylation of phytocannabinoids, thereby generating pharmacologically active CBs, and these conditions may differ for each cannabis cultivar.

Conclusion : Chemical consistency and potency due to active compounds are in turn important in producing consistent and reliable medical cannabis extracts and their derivatives. These processes must be subject to higher levels of scientific rigor as the patient population around the world are seeking the help of such extracts for various clinical conditions, and as medical cannabis industry is receiving acceptance in various countries.

Keywords : cannabinoid receptors; decarboxylation; extraction; medical cannabis; phytocannabinoids



Numerous international and United Nations regulations served to control cannabis, and only in the recent years has the medicinal use of cannabis been again realized. Accordingly, the regulations are being reviewed and/or modified in this regard, thus allowing researchers to investigate the myriad of natural compounds present in cannabis in academic laboratories.1,2,# Within the cannabis plant, at least 568 compounds have been identified to date, of which *120 are phytocannabinoids. 3,4 Phytocannabinoids are biosynthesized from phenolic precursors in the cannabis plant, and several of these molecules carry a carboxylic acid moiety; major phytocannabinoid acids include D9-tetrahydrocannabinolic acid (D9 THCA, 1), cannabidiolic acid (CBDA, 3), and cannabigerolic acid (CBGA, 5). Synthetic as well as plant derived cannabinoids bind to cannabinoid (CB) receptors 1 and 2 (CB1 and CB2, respectively) in the central and peripheral tissues, and modulate these receptor responses for subsequent physiological effects.5–7

Acidic phytocannabinoids such as D9-THCA exhibit poor potency at CB receptors (cannabimimetic activity), whereas the decarboxylated phytocannabinoids such as D9 tetrahydrocannabinol (D9-THC), also referred to as neutral phytocannabinoids, exhibit high affinities and physiological activities.8–11 The decarboxylation step and the consistency of such decarboxylation are important to achieve reliable pharmacological effects when medical cannabis and its derivatives are used for their therapeutic efficacy.12–14 Patients typically smoke or vaporize the whole plant, or ingest the extract as edibles and infused edible oils. New industries are being fostered, focusing on medical cannabis extracts and their medical uses. In this context, cannabis extracts have become very popular in the recent months, including receiving U.S. Food and Drug Administration (FDA) approvals. For example, just a few months ago, Epidiolex, a cannabisextracted drug with >98% cannabidiol (CBD) and <0.5% of D9-THC, was approved by U.S. FDA for the treatment of intractable epilepsy including for pediatric use.15–17

There are conventional and domestic methods described for cannabis extraction such as ethanol extraction, maceration, butane extraction, and quick-wash alcohol extraction. Recently reported extraction methods include ultrasound-assisted extraction (UAE) and supercritical fluid extraction (SFE).18–21 Each method carries advantages and disadvantages depending on the compounds to be extracted, duration of extraction, temperature, and solvent, if any. It is typically desirable to use a solvent that solubilizes and carries compounds from the plant, and the temperature for extraction should minimize the loss of thermally labile groups or unwanted chemical transformations.

Soxhlet extraction involves continually extracting soluble phytochemicals from the plant under refluxing conditions of the solvent, typically ethanol. Soxhlet extraction may present few challenges such as duration of extraction, efficiency, and postextraction processing. Microwave-assisted extraction (MAE) employs microwaves to assist in the extraction of compounds from cannabis at elevated temperatures and pressures.22–24 The instantaneous energy transfer from solvent to biomass leads to a rapid increase in the temperature, and one can reach temperatures higher than the boiling point of the solvent if the pressure is contained.25 MAE offers additional advantages such as shorter extraction and reaction times, smaller quantities of solvent, and reproducibility.25 While all these methods help in extracting phytocannabinoids and other compounds from cannabis, activation of the extracts through the decarboxylation of phytocannabinoids remains a unique challenge.

Decarboxylation of acidic phytocannabinoids could occur in an open or closed reactor. In an open reactor, decarboxylation of CBs has been demonstrated to occur at 37C and 60C after exposure for several hours or at 100C for 60 min; in a closed reactor, the reaction could reach completion at 200C in just 3min.26 A disadvantage of using an open reactor for decarboxylation is that there is no agreeable temperature under atmospheric conditions at which efficient decarboxylation of the acidic CBs can occur without simultaneous evaporation of the solvent, along with any volatile compounds.26 Every phytocannabinoid carboxylate would have a different optimal condition for decarboxylation (Fig. 1), thus various medical cannabis cultivars with various chemical compositions would require different conditions to achieve complete decarboxylation of all phytocannabinoids.

A wide range of concentrations of D9-THC and CBD and their carboxylic acid precursors, D9-THCA and CBDA, are present in commercial medical cannabis products.27,28 If consumed, patients would be exposed to varying quantities of active phytocannabinoids leading to inconsistent physiological response. This could vary from batch to batch of the same product, as is often the case with plant-based products, providing highly uncertain and variable physiological effects. Due to the significant and renewed interest inmedical cannabis extracts, their associated chemistry and biological activities, we undertook a comprehensive investigation to compare various chemical extraction protocols using three medical cannabis cultivars commercially sold in Canada. It is also important to note that medical cannabis is not a single substance, and is essentially classified into hundreds of varieties based on the composition of its phytocannabinoids. In this study, we explored three varieties of medical cannabis cultivars (balanced D9- THC/CBD, high CBD, and high D9-THC) with chemical distinction investigating their phytocannabinoid profiles, compared the efficiencies of extractions, chemical compositions, decarboxylation efficiencies of phytocannabinoid acids, and discussed their relevance to CB receptor responses. This study paves the way for further investigations into medical cannabis, relevant CBs, and the corresponding receptor responses.