CSCW: The Analysis of 21 Cannabinoids by LC-MS

Significance of Analyzing Cannabinoids

Accurate measurement of cannabinoid content in hemp and cannabis products is essential for regulatory compliance, consumer safety, and research advancement. As new minor cannabinoids are discovered, sensitive and selective analytical methods are required to support potency testing and product labeling.

Objectives and Study Overview

This work aimed to develop a rapid liquid chromatography mass spectrometry method capable of quantifying 21 cannabinoids in a nine minute cycle. The study focused on achieving baseline separation of critical isobaric pairs while maintaining high throughput and the ability to expand the analyte panel as new cannabinoids emerge.

Used Instrumentation

• UHPLC system equipped with a Raptor ARC-18 analytical column, 150 mm x 2.1 mm, 2.7 μm particles, 90 angstrom pore size and matching guard column
• ESI source operating in positive and negative polarity
• Single ion monitoring acquisition mode on a mass spectrometer

Methodology

The separation employed an isocratic mobile phase composition of water with 0.1 percent formic acid and 12 mM ammonium formate mixed with a 50:50 acetonitrile methanol blend containing 0.1 percent formic acid. The flow rate was held at 0.5 mL per minute, column temperature at 30 degrees Celsius, injection volume at 2 microliters, and sample concentration at 500 nanograms per milliliter. Five groups of isobaric cannabinoids sharing identical mass were chromatographically resolved based on slight retention differences.

Main Results and Discussion

The method achieved baseline separation for all 21 cannabinoids within nine minutes. Key isobaric groups included:

• Group 1 : Cannabidivarin, tetrahydrocannabivarin and cannabichromevarin
• Group 2 : Cannabidiphorol and tetrahydrocannabiphorol
• Group 3 : Cannabidiolic acid, tetrahydrocannabinolic acid A, cannabichromenic acid and cannabicyclolic acid
• Group 4 : Cannabidivarinic acid and tetrahydrocannabivarinic acid
• Group 5 : Cannabidiol, delta 9 tetrahydrocannabinol, delta 8 tetrahydrocannabinol, cannabicyclol, cannabichromene and cannabicitran

Intensity profiles confirmed selective detection via single ion monitoring. The isocratic approach simplified method setup and allowed straightforward panel expansion.

Benefits and Practical Applications

• High throughput analysis supports testing laboratories with rapid sample turn around
• Mass selective detection enhances sensitivity compared to UV based methods and removes need to resolve non isobaric compounds
• Method flexibility accommodates addition of new cannabinoids with minimal reoptimization

Future Trends and Potential Uses

As novel cannabinoids continue to be identified, the panel can be extended by targeting additional m/z values and optimizing retention for new isobaric pairs. Advances in high resolution MS and automated sample preparation will further improve sensitivity, accuracy and laboratory productivity.

Conclusion

A nine minute LC MS method was developed for simultaneous quantitation of 21 cannabinoids, achieving reliable separation of critical isobaric groups. This approach delivers enhanced sensitivity, throughput and adaptability for evolving potency testing requirements.

Reference

1. Linciano P et al Journal of Natural Products 2020 83 88-98
2. Citti C et al Data in Brief 2019 26 1-16