PSIII-1 - Effect of cannabidiol extraction on the cell wall carbohydrate composition of hemp inflorescences

Abstract: Since the 2018 Farm Bill, US growers have been allowed to cultivate hemp (Cannabis sativa L.), prompting interest in exploring the use of hemp products in ruminant diets. Extraction of cannabidiol (CBD) oil from hemp biomass generates spent inflorescent material that could be upcycled as a feedstuff, yet little is known about its fiber composition. A recent study measured in vitro true digestibility and fermentation parameters of multiple hemp cultivars, some intact and some CBD-extracted (Altman et al., 2022). Both categories of hemp inflorescence material had similar or greater total digestible nutrients (TDN) than an alfalfa cube control, supporting hemp’s potential as a feedstuff. However, comparisons between non-extracted and CBD-extracted samples could not be made, and the fiber content was only described as neutral detergent fiber (NDF). Fiber composition can significantly impact ruminal digestibility, and in-depth characterizations of cell wall carbohydrate composition can provide further insight into the digestibility of a specific plant-based feed. Therefore, in the present study, we examined the effect of CBD oil extraction on the cell wall carbohydrate composition of the inflorescent material of two hemp cultivars. Intact (Biomass) and CBD-extracted (Spent) inflorescent material from two hemp cultivars, Mary and Zelios, was analyzed. The samples’ cell wall monosaccharides were released via two complementary acid hydrolyses, methanolysis and Saeman (sulfuric acid) hydrolysis, and separated and quantified using high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD). Additionally, the carbohydrate linkage patterns were assessed by generating partially methylated alditol acetates (PMAAs). Student’s unpaired t-tests were run in GraphPad Prism, and no differences (p>0.05) in monosaccharide concentrations based upon treatment (CBD extraction) or cultivar were found with either hydrolysis method except for the methanolysis glucose concentration between the biomass forms of the two cultivars where Mary Biomass possessed more glucose than Zelios Biomass (p < 0.001). In the Saeman hydrolysates, glucose was the dominant monosaccharide (38.75 ± 1.44 molar %), indicating a large amount of cellulose, supported by the presence of 1,4-Glcp in the linkage analysis. Linkage analysis also revealed the presence of xyloglucans and rhamnogalacturonan Type 1 pectins in all materials. The data show no effect of CBD oil extraction on the monosaccharide composition, suggesting that the extraction process leaves the fiber components mostly intact, but there may be differences in glucose concentration (and consequently cellulose content) between cultivars of hemp. Future studies may benefit more from investigating the impact of hemp cultivar on fiber digestibility rather than impact of the CBD extraction process.