Compliance Testing of Hemp (Cannabis sativa L.) Cultivars for Total Delta-9 THC and Total CBD Using Gas Chromatography with Flame Ionization Detection

The United States Agriculture Improvement Act passed in December of 2018 legalized the growing of Cannabis sativa containing not more than 0.3% total Delta-9 tetrahydrocannabinol (THC) in the country. While Cannabis sativa has been cultivated for hundreds of years, the illegal status of the plant in the United States, and elsewhere, has hindered the development of plant cultivars that meet this legal definition. To assess sampling strategies, and conformance to the THC limit, 14 cultivars of hemp were grown and tested by using gas chromatography with flame ionization detection for total delta-9 THC and total cannabidiol (CBD) during 2020, 2021 and 2022. Each year, samples of fresh plant material were collected from each cultivar weekly, beginning in mid-August and ending in late October, to examine the rate of increase in THC and CBD for different cultivars and select individual plants. The sampling demonstrated that both CBD and THC increase rapidly over a 1-2-week time frame with maximum concentrations (about 16% and 0.6%, respectively) around late September to early October. The testing of individual plants on the same day for select cultivars showed that while the ratio of CBD to THC remains constant (about 20:1 in compliant hemp) during the growing season, the individual plants are highly variable in concentration. Whereas previous studies have shown cultivar-dependent variability in THC production, this study demonstrated a novel plant-to-plant variability in the levels of THC within the same hemp cultivar. Understanding variability within and between hemp cultivars is useful to determine field sampling strategies and to assess the risk of crop embargoes to growers by compliance regulators.