Separation and non-separation methods for the analysis of cannabinoids in Cannabis sativa L.

Investigating the effect of polarity of stationary and mobile phases on retention of cannabinoids in normal phase liquid chromatography

“…Initially, the CBD-rich sample, extracted from non-psychoactive cannabis (G), was characterized by means of RPLC [ 47 , 48 ], for the identification and quantification of the main cannabinoids. This information will be used later on to compare RP and NP chromatography.…”

“…The same experimental conditions have been applied to the separation of a real sample (“ Sample preparation ”) in order to have some insights about the possible effects on the separation coming from the presence of other compounds in the real matrix, on the one hand, and the relative abundance of cannabinoids, on the other. Figure 6 reports cromatograms measured on the entire set of columns with Hept/IPA and Hept/EtOH as mobile phases for the cannabis extract G. This sample has been previously fully characterized using a fully validated analytical method [ 48 ]. In most cases the five cannabinoids are well resolved and can be easily identified.…”

NMR Spectroscopy Applied to the Metabolic Analysis of Natural Extracts of Cannabis sativa

“…It is worth noting that the NMR technique has been proven to be a reliable and powerful tool for the quantification of different natural products and, especially in recent years, has also been successfully applied to the quantification of CBD and other cannabinoids directly on hemp extracts coming from different cultivars using both the 1 H and 13 C qNMR methodologies. [ 40 , 44 , 68 , 69 , 70 , 71 ]. Quantitative 1 H-NMR is the most widely used method in the quantification of natural extracts and has been shown to have a good level of accuracy and reproducibility.…”

“…For each 1 H NMR experiment, 128 FIDs were acquired using a spectral width of 14.00 ppm and a relaxation delay of 5.0 s. The 1D 13 C-{ 1 H} NMR spectra were recorded with proton broad-band decoupling, collecting 8K FIDs using a SW of 250.00 ppm and a relaxation delay of 5.0 s. For an accurate quantitative analysis of the metabolites present in the complex mixture, it was necessary to calibrate both the 90° pulses on the monitored nuclei ( 1 H and 13 C) and the T1 spin-lattice relaxation time. The T1 relaxation time was estimated by using the conventional inversion recovery experiment (10 increments from 0.5 ms to 30.0 s for 1 H, and 24 increments from 0.1 ms to 300.0 s for 13 C) [ 44 ]. The 1 H quantitative NMR (qNMR) spectra were recorded using the same acquisition parameters described before but with a relaxation delay of 20.0 s. Instead, for 13 C qNMR, quantification experiments ( zgig Bruker pulse sequence) were performed, collecting 4000 FIDs using a SW of 250.00 ppm, a relaxation delay of 160.0 s, and an acquisition time of 10.0 s.…”

Antiseizure Effects of Fully Characterized Non-Psychoactive Cannabis sativa L. Extracts in the Repeated 6-Hz Corneal Stimulation Test

“…Specifically, we aimed to test the in vivo antiseizure effect of olive oil extracts from hemp inflorescences, which were fully characterized for their content of both cannabinoids and terpenes. Even though it was challenging to study the chemical composition of plant extracts, in view of their complex chemical composition, the full characterization of both cannabinoids and terpenes was made possible and reliable thanks to specific and fully validated analytical methods based on high-performance liquid chromatography (HPLC) and gas chromatography (GC) [20][21][22][23][24]. The influence of the compounds found in the extracts, specifically terpenes in combination with cannabinoids, on the experimental results obtained after the administration of the oil extracts and pure CBD in mice is illustrated in detail.…”