Simple chemical tests to identify Cannabis derivatives: Redefinition of parameters and analysis of concepts

The chemical identification of Cannabis is commonly carried out using the Duquenois-Levine (DL) colorimetric test. On the other hand, its active substances called cannabinoids are differentiated by thin-layer chromatography (TLC). This work aims to optimize parameters of these two chemical tests using different samples of Cannabis in natura and previously heated (decarboxylated), as well as their isolated bioactive and possible false positives. The efficiency of the DL test without using ether and aliphatic aldehyde was evaluated, comparing the removal or not of the solid sample from the reaction medium after applying different concentrations of the vanillin ethanolic solution. The chemical properties of different cannabinoids and solvents were estimated and correlated with the TLC retention factors. DL tests applied directly to the plant matrix did not show the expected color, even using a high concentration of vanillin. However, obtaining ethanolic extracts from the samples using low vanillin concentration was sufficient without detecting false positives described in the literature. Cannabinoids with high dipole moment ( $\mu$ ) were poorly eluted in TLC, indicating a great interaction with the stationary phase. Their identifications could be conducted based on their distinct lipophilic characteristics ( $\log P_{\mathrm{OW}}$ ) and the choice of a more polar solvent mix ( ${\mu }_{\mathrm{mix}}$ ). It is concluded that the DL test can be conducted with reagents of less toxicity, but it is necessary to remove the plant matrix from the reaction medium. The correlation of the TLC results with the chemical properties of cannabinoids and solvents was consistent and can be extrapolated for more complex analyses.