How to overcome co-elution challenges in cannabinoid chromatography using tandem mass spectrometry?
Research Poster Health & Life Sciences 2025 Graduate ExhibitionPresentation by Sujata Chatterjee
Exhibition Number 81
Abstract
In this collaborative study with Penn State plant scientists, we investigated how different varieties and soil nutrients affect cannabinoid concentration in Cannabis sativa (hemp) plant. The hemp inflorescence accumulates tetrahydrocannabinol (THC), cannabidiol (CBD) and their metabolites, most of which are poorly resolved on chromatographic systems due to similar chemistries. Thus, the first objective of this research work was to develop a simple approach to successfully resolve individual cannabinoids and subsequently measure them in a plant matrix. To achieve this, we first employed reverse phase liquid chromatography to identify and separate standard mixtures of 8 cannabinoid neutrals and 6 cannabinoid acids. Amongst the most difficult to resolve were CBD and cannabigerol(CBG). However, since they had slightly unique fragmentation patterns, tandem mass spectrometry (MS/MS) overcame this limitation by offering extremely selective mass to charge ratios through Multiple Reaction Monitoring (MRM). Similarly, unique fragments of each cannabinoid were optimized using specific cone voltage and collision energies on the Waters Xevo TQ-S mass spectrometer, which led to successful resolution of 14 individual cannabinoids within 17 minutes. To put our method to test, we analyzed a hemp inflorescence extract, and our method shows excellent sensitivity, linearity, and resolution with minimal matrix interference. Next, we plan to validate the method and report both individual and total cannabinoids on plant dry weight basis. Upon knowledge transfer to horticultural or cannabis testing labs, this analytical method can aid in better agricultural decisions like selection of strain and nitrogen application for optimum cannabinoid yield.
Importance
Cannabinoids help plants withstand stress conditions like disease and nutrient deficiency and serve as active ingredients in healing formulations, skincare and alternative medicine, comprising a substantial market share. Total cannabinoid yield (grams of CBD/grams of plant dry mass) is crucial for growers as it is related to its commercial value. Additionally, individual components (such as cannabichromene, cannabigerol, and cannabivaric acid, to name a few) are of increasing interest to the pharmaceutical industry due to their unique therapeutic applications. Since this method measures both individual and total cannabinoids, it can help plant scientists and hemp growers optimize their farming inputs to maximize cannabinoid yield, leading to smarter resource utilization and less wastage on the field.