Comparing the Utility of Cannabidiol Quantitation Methods for Use in High-Throughput In Vitro Assays

In recent years, isolated phytocannabinoids have received significant attention for their therapeutic potential, showing diverse pharmacological effects in vitro and in preclinical studies. However, in vitro drug efficacy and drug development studies are challenging to perform as accurate quantification of phytocannabinoids is required to generate dose response curves and pharmacokinetic profiles. While assessment of free cannabinoids is straightforward, assessing small quantities in cell culture media or as part of a formulation is difficult due to the low solubility of cannabinoids in aqueous medium and high propensity for adsorbing to plastics. Therefore, to pursue a phytocannabinoid drug development program, one of the major obstacles to overcome is selecting an appropriate analytical method that provides both accuracy and efficiency for cannabinoid quantitation. To address this challenge, three methods for quantifying cannabinoids are compared: high performance liquid chromatography (HPLC), UV/vis Spectrophotometry, and colorimetric analysis using Fast Blue B Salt (FBBS). Each method demonstrates advantages and limitations, and a comprehensive understanding of their utility in different experimental workflows is necessary for advancing the study of cannabinoids as therapeutic agents.