Study formulation
The first step to determine compounds other than CBD and CBG to include in the study formulation was an environmental scan of compounds in commercially available recovery-based products. The next step was a comprehensive search of the published literature to collect scientific evidence on the recovery-promoting properties of compounds identified in the environmental scan. Results from published studies were documented, as were aspects of study methods that could impact the interpretation of results, including species, description of sample for studies of humans (e.g. exercise-trained or not), study design (e.g. clinical trial, experimental study of mechanism of action), sample size, dose of compound(s), description of exercise manipulation, reported safety signals, and outcome measures. Compounds were selected for inclusion in the study formulation based on strength of evidence regarding safety in humans and recovery-enhancing properties.The active formulation was a powder in a 7.0 g single-serving packet that delivered 35 mg CBD, 50 mg CBG, 25 mg BCP, 3.8 g BCAAs, and 420 mg magnesium citrate. Because CBD and CBG are highly lipophilic molecules that do not dissolve well in water, the active formulation included HydroBond CBD™ and Hydrobond CBG™ water-soluble powders. These powders consisted of 20% of the given cannabinoid and 80% acacia gum and were manufactured into a nano-emulsion and spray dried to form a powder. The resulting powders were readily dispersible in liquid with no oily surfactants or bitter taste. Participants consumed 2 doses of study formulation per day; thus, the total daily consumption of active formulation was 70 mg CBD, 100 mg CBG, 50 mg BCP, 7.6 g BCAAs, and 840 mg magnesium citrate. Dosing was informed by federal agencies’ recommended daily limits, published data on tolerated doses in animals and effects on recovery in humans, and scans of the commercial market [The placebo formulation was a powder in a 7.0 g single-serving packet that contained maltodextrin. To ensure similar flavor and color and thus maintain blinding, both active and placebo formulations contained passionfruit coconut natural flavor, Stevia, and beta carotene. Study personnel and participants were blinded to formulation assignment. Each participant only received one formulation in this parallel groups study design, thereby reducing the risk of direct comparison between active and placebo. Several steps were undertaken to ensure adequate blinding of study formulation. First, the study formulation was dispensed in single-serving sachets, with blinded labels. Second, the formulation contained water-soluble cannabinoids made from high purity distillates/isolates, not full spectrum extracts, thus limiting the distinctive smell and taste of cannabinoids. Third, flavoring, which included bitter blockers, was designed to mask the cannabinoids and provide a pleasant taste. Fourth, members of the research team and the team that formulated the active and placebo study formulations evaluated both formulations to ensure similarity, and no obvious visual features differed between formulations.Participants were instructed to mix the powder with 16 oz. of water, and to consume orally. Participants were also instructed to consume the study formulation at 8 PM (±1 hour) the night of the first study visit, and then at 8 AM and 8 PM (±1 hour) every day until their final follow-up visit. The last dose occurred at 8 AM (±1 hour) immediately prior to the 72-hour post-DOMS study visit. To improve the bioavailability of cannabinoids, participants were recommended to consume the study formulation approximately 1 hour after completing a meal [CBD and CBG were hemp-derived (derived from hemp containing no more than 0.3 percent THC on a dry weight basis) and thus were not controlled substances under U.S. federal law. The active and placebo powders were manufactured for the sponsor by a contract manufacturer according to current Good Manufacturing Practice (cGMP) 21 CFR part 111. Potency of cannabinoids (e.g. CBD, CBG) and within the final product was verified by both Botanacor (Denver, CO) and Eurofins (Madison, WI). Testing of BCP, magnesium citrate, and BCAAs was conducted at Eurofins (Madison, WI), and testing of contaminants (e.g. heavy metals, mycotoxins, pesticides, residual solvents, microbiology) was conducted by ACS Laboratory (Sun City, FL).