Regulatory sampling of industrial hemp plant samples (Cannabis sativa L.) using UPLC-MS/MS method for detection and quantification of twelve cannabinoids

Abstract

Background In 2018, the Farm Bill mandated the United States Department of Agriculture to develop regulations governing the cultivation, processing, and marketing of industrial hemp. Industrial hemp is defined as Cannabis sativa L. with a total Δ-9-tetrahydrocannabinol (Δ-9-THC) content ≤0.3%. Therefore, for hemp to become an agricultural commodity, it is important to regulate production by developing standard methods for sampling and testing of the plant material. Methods An ultra-performance liquid chromatography-tandem mass spectrometry analytical method for the quantification of twelve cannabinoids was developed. The method was applied to a regulatory sampling trial of three hemp varieties cultivated for cannabidiol (CBD) production. Two samples were taken from 28 plants with one sample being flower only while the other was a composite sample that included flowers, leaves, and stems. Results The assay method was validated for specificity, range, repeatability, reproducibility, and recovery in accordance with all applicable standards for analytical methods. The results of the regulatory study indicated a significant decrease in the concentration of total Δ-9-THC and total CBD of 0.09% and 1.32%, respectively, between a flower only and a composite sample. Conclusions There are many factors that may influence reported total Δ-9-THC content in industrial hemp. A robust analytical method was developed to analyze hemp samples in a trial regulatory study. The results indicate that the way hemp is sampled and analyzed may influence the legality of a crop, which could have negative economic and legal consequences.

Response of essential oil hemp (Cannabis sativa L.) growth, biomass, and cannabinoid profiles to varying fertigation rates

“…Dried samples were ground into a fine powder using a coffee grinder and stored in 100 mL glass vials. Detailed extraction and quantification of cannabinoids methods can be found in Berthold et al [30]. Ground samples were weighed, and cannabinoids were extracted by adding a solution of methanol and water (95:5, v/v) acidified with 0.005% formic acid at a 1:100 w/v plant material to solvent concentration ratio.…”

“…Supernatant was serial diluted using extraction solvent until the sample quantification fell within quantification range. Quantification of cannabinoids was conducted using a Waters I-Class Acquity UPLC (Milford, MA, U. S.) coupled with a Waters Xevo TQ-S Micro™ triple-quadrupole mass spectrometer (MS/MS) [30]. Furthermore, mass spectrometry was used for detection, no wavelengths are involved just mass transitions.…”

“…Furthermore, mass spectrometry was used for detection, no wavelengths are involved just mass transitions. The mass transitions are available in Berthold et al [30]. Raw cannabinoid quantification and standard curve data are presented in S1 File.…”

How do pinching and plant density affect industrial hemp produced for cannabinoids in open field conditions?

“…Harvest index was calculated as the ratio of flower yield to shoot biomass. Stripped flowers were then ground into fine powder using a small coffee grinder for cannabinoid analysis following the ultra-performance liquid chromatography-tanden mass spectrometry method reported by Berthold et al (2020). Three analytical replicates were used for each sample.…”

Industrial Hemp Clone Selection Method under LED Smart Farm Condition Based on CBD Production per Cubic Meter

“…Cannabis breeders combine several genes to develop economically valuable fiber, seed, and medicinal hemp [28]. The 'Cherry Wine' variety used in this experiment is a hybrid of The Wife × Charlotte's Cherries and is a CBD-only variety with a total THC content of less than 0.3% [27]. Initially, wild cannabis provided beneficial seeds, fibers, and drugs for humans.…”