New Forensic Methods to Accurately Determine THC in Seized Cannabis

NIJ-funded researchers address the need for simple, cost-effective ways to differentiate hemp from marijuana.

Date Published February 13, 2024

The Controlled Substance Act of 1970 classified the plant cannabis, which was historically classified as either marijuana or hemp, as an illegal drug, a Schedule I controlled substance with a high potential for abuse and no FDA-approved medical use in the United States. For almost 50 years, hundreds of thousands of people were arrested and imprisoned for possessing it.

The Farm Bill of 2018 changed this straightforward classification of cannabis. The bill legalized the form of cannabis classified as hemp, while the form classified as marijuana remained illegal. The law left how to tell the difference up to law enforcement and forensic laboratories, which has proven difficult, time consuming, and expensive.

Cannabis is considered marijuana if it is greater than 0.3% THC and hemp if there is less than that.

Scientists use the amount of THC (one of the psychoactive components of cannabis) present in a cannabis plant to differentiate hemp from marijuana. [1] The Farm Bill defines hemp as cannabis with 0.3% or less total THC. A cannabis plant with more than 0.3% THC is considered marijuana and remains on the Schedule I substance list (Figure 1).

The fact that many states have decriminalized the use of marijuana further complicates cannabis regulation. Currently, the medical use of marijuana is legal in 37 states and the District of Columbia, and the recreational use of marijuana is legal in 19 states and the District of Columbia. In the remaining states, however, marijuana is still considered unlawful and must be distinguished from hemp. [2] Federal law continues to consider marijuana illegal.

As a result of the Farm Bill, forensic laboratories must measure the exact amount of THC in seized evidence to differentiate hemp from marijuana. In 2022, more than 10% of all submissions to crime labs were to determine marijuana versus hemp.

Thus, a new problem emerged for already backlogged crime labs. H ow could labs perform such precise measurements quickly and easily when few labs had the personnel, instrumentation, and protocols to do so?

Is It Hemp or Marijuana?

Two NIJ-supported labs addressed this problem using different types of mass spectrometry that can measure the exact amount of THC present in a sample: gas chromatography-mass spectrometry (GC-MS) and direct analysis in real time-high-resolution mass spectrometry (DART-HRMS).

Historically, most forensic laboratories have used qualitative tests for seized cannabis samples to distinguish hemp from marijuana. Labs could confirm micro- and macroscopic plant features, screen for THC through a colorimetric test (which changes color in its presence), or test for the presence of THC through various gas chromatography techniques or thin layer chromatography (which separate the different components of cannabis). But none of these tests can measure the exact amount of THC present in a sample.

Dr. Walter Brent Wilson’s team at the National Institute of Standards and Technology used GC-MS to develop a simple, robust, and cost-effective method to distinguish hemp from marijuana for local, state, and federal forensic laboratories. They are expanding their work to include edibles. With support from NIJ, their research:

Dr. Rabi Musah’s lab from the University at Albany, State University of New York, approached the problem differently. The lab used DART-HRMS to quantify the amount of THC found in complex materials that are often difficult to study, such as edibles, beverages, and plant materials. This method can be performed directly on the sample with minimal to no sample pre-treatment. Their project:

The University of Albany team showed that they can use DART-HRMS to rapidly detect THC and other cannabis-related molecules in baked goods, candies, beverages, and plant materials with minimal pre-treatment steps. They also identified the limits of using DART-HRMS to detect THC in different types of samples.

Impact on Forensic Laboratories

Dr. Musah anticipates that the increased speed of using DART-HRMS to detect THC in cannabis samples could reduce sample testing backlogs and chemical reagent costs and streamline sample analysis protocols.

Dr. Wilson’s project created standard operating procedures for quantifying THC in federal, state, and local forensic laboratories. His research team has shared findings through webinars and publications and created training models for the Montgomery County Police Department and Maryland State Police crime labs on using GC-MS to quantify THC.

About This Article

The work described in this article was supported by NIJ inter-agency agreement number DJO-NIJ-20-RO-0009, awarded to the National Institute of Standards and Technology, and by NIJ grant number 2019-DU-BX-0026 , awarded to the University of Albany, State University of New York. This article is based on the reports from those awards: “ Accurate THC Determinations on Seized Cannabis Samples for Forensic Laboratories ” (pdf, 26 pages) by Walter Brent Wilson, Ph.D. and “ Research to Develop Validated Methods for THC Quantification in Complex Matrices by High-resolution DART-MS-Focus on Edibles and Plant Materials ,” (pdf, 18 pages) by Rabi A. Musah, Ph.D.

Sidebar: How is Mass Spectrometry Used in Cannabis Analyses?

Mass spectrometers allow scientists to calculate the exact molecular weight of a substance and identify known and unknown compounds.

One type of mass spectrometry, gas chromatography-mass spectrometry (or GC-MS), combines the features of gas-chromatography and mass spectrometry to identify different substances within a test sample. Applications of GC-MS include drug detection, fire investigation, environmental analysis, explosives investigation, food and flavor analysis, and identification of unknown samples.

Another type of mass spectrometry, direct analysis in real time-high-resolution mass spectrometry (or DART-HRMS), combines the features of direct analysis in real time with high-resolution orbitrap mass spectrometry to identify different substances within a test sample. Scientists can use DART-HRMS to obtain exact mass measurements rapidly with high-resolution mass spectrometers without having to use a separation step. Applications of DART mass spectrometry include pharmaceutical investigations, forensic studies, quality control, and environmental studies.

Notes

[note 1] THC refers to Δ9-tetrahydrocannabinol, which is the major psychoactive component in cannabis. THCA is tetrahydrocannabinolic acid, and it is the most abundant non-psychoactive cannabinoid in cannabis. Here, total THC refers to Δ9-tetrahydrocannabinol plus THCA.

In scientific terms, to find the “Δ-9 tetrahydrocannabinol concentration … on a dry weight basis” as stated in the statute, it is necessary to consider both the Δ9-THC concentration as well as the concentration of THCA.

The 0.3% threshold was chosen based on the outdated work of the Canadian scientist Ernest Small, who found the threshold useful based on years of plant use research in the 1970s and not based on abuse or intoxication patterns. For the original, see E. Small and A. Cronquist, “A practical and natural taxonomy for Cannabis,” Taxon 25 no. 4 (1976): 405-435, https://doi.org/10.2307/1220524.

[note 3] Full-scan mode scans from mass-to-charge ratio (m/z) 50 to 350 ion while single-ion monitoring mode quantitates at 299 ion.