This study used nanofluidic protein posttranslational modification (PTM) profiling to measure the effects of six cannabidiol (CBD) oils and isolated CBD on the signaling pathways of a cultured SH-SY5Y neuronal cell line. Chemical composition analysis revealed that all CBD oils met the label claims and legal regulatory limit regarding the CBD and tetrahydrocannabinol (THC) contents, respectively. Isolated CBD was cytotoxic, with an effective concentration (EC50) of 40 µM. In contrast, the CBD oils had no effect on cell viability at CBD concentrations exceeding 1.2 mM. Interestingly, only an unadulterated CBD oil had strong and statistically significant suppressive effects on the pI3K/Akt/mTOR signaling pathway with an EC50 value of 143 µM and a slow-acting timescale requiring hours. Systematic profiling of twenty-six proteins, which served as biomarkers for nine signaling pathways, revealed that the unadulterated CBD oil downregulated seven signaling pathways but had no measurable effect on the other two signaling pathways. The remaining CBD oils, which were adulterated, and isolated CBD had weak, variable, or undetectable effects on neuronal signaling pathways. Our data clearly showed that adulteration diminished the biological activities of CBD oils. In addition, nanofluidic protein PTM profiling provided a robust means for potency assessment of CBD oils.
KEYWORDS: adulteration, cannabidiol, capillary isoelectric focusing, nanofluidic proteomics, neuronal signaling pathways, potency assessment, protein posttranslational modification
- PMID: 32019055
- DOI: 10.3390/nu12020357
The authors declare no conflicts of interest. C.B., M.W., and D.K.H. are employees of dōTERRA International. Currently, dōTERRA International does not produce or sell hemp extracts or cannabinoid products. D.K.H. and C.B. participated in the initial conceptualization of the project. C.B. and M.W. analyzed the chemical composition of CBD oils purchased from the open market using HPLC and GC-MS methods. Data collected by C.B. and M.W. were compared side by side with those collected independently by the Aromatic Plant Research Center and Botanacor.