cannabidiol and cannabielsoin

The reported characteristics of cannabidiol (CBD) have encouraged significant growth in commercial CBD products. There is limited information on the stability of CBD and some researchers have noted significant reductions of CBD in products. In this study, the chemical profiles of plant-based and chemically synthesized CBD in a prototype e-liquid formulation were assessed during 4 weeks of storage under varying conditions. Samples were analysed on days 1, 8, 15, 22, and 29 by untargeted analysis using ultra-high performance liquid chromatography-trapped ion mobility-time-of-flight mass spectrometry (UHPLC-TIMS-TOF-MS). On day 1, analysis of plant-based and synthetic CBD formulations showed small differences in their composition, with plant-based CBD e-liquid containing trace levels of a higher number of phytocannabinoid-related impurities. Storage for 4 weeks under stress (40 °C, 75% relative humidity, dark) and ambient (25 °C, 60% relative humidity, daylight) conditions led to increases in the number and abundance of cannabinoid-related degradation products, including cannabielsoin (CBE) and CBD-hydroxyquinone (HU-331), which are products of the oxidation of CBD, and other unidentified cannabinoid-related compounds. The unidentified cannabinoid-related compounds were probed by accurate mass measurement and MS

Topics: Cannabidiol; Cannabinoids; Chromatography, High Pressure Liquid

In this report, we revise the structure for a previously reported synthetic product proposed to be the 1R,2S-cannabidiol epoxide and reassign it as cannabielsoin using anisotropic NMR and synthetic chemistry methods. These results provide a direct link to the first known biological target and function of cannabielsoin.

Topics: Anisotropy; beta Catenin; Cannabidiol; Magnetic Resonance Spectroscopy; Molecular Conformation; Wnt Proteins

Cannabielsoin (CBE) was identified as a novel metabolite of cannabidiol (CBD) in the guinea pig in vivo and in vitro. Its formation by liver microsomes of guinea pigs needed NADPH and molecular oxygen, and was inhibited with SKF 525-A, metyrapone and alpha-naphthoflavone, indicating participation of cytochrome P-450 (P-450). The CBE-forming activity was highest in guinea pigs, followed by mice, rabbits and rats. In the rat, sex difference was found in the CBE formation (male greater than female). CBD monomethylether (CBDM) was also biotransformed to CBE monomethylether (CBEM) in the guinea pig in vivo and in vitro. When CBD dimethylether (CBDD) was employed as substrate, 1S,2R-epoxy-CBDD was identified. The results suggest that CBD and CBDM are biotransformed by P-450 to CBE-type metabolites via 1S,2R-epoxides. In pharmacological studies using mice, CBDD and 1S,2R-epoxy-CBD-2',6'-diacetate produced hypothermia, and CBD, CBDM and CBEM prolonged pentobarbital-induced sleep. Moreover, 1S,2R-epoxy-CBD-2',6'-diacetate was examined in the Ames test, but had no mutagenicity.

Topics: Animals; Body Temperature; Cannabidiol; Cannabinoids; Catalepsy; Chromatography, Gas; Chromatography, High Pressure Liquid; Enzyme Induction; Female; Gas Chromatography-Mass Spectrometry; Guinea Pigs; In Vitro Techniques; Liver; Male; Mice; Mutagenicity Tests; Pentobarbital; Rabbits; Rats; Sleep

Oxidative metabolism of cannabidiol monomethyl ether (CBDM), one of the components of marihuana, was studied in the guinea pig. Cannabielsoin monomethyl ether (CBEM) was found to be formed with hepatic microsomes by gas chromatography-mass spectrometry (GC-MS). Experiments using various modifiers of enzymatic reaction suggested that, as in the case of cannabielsoin (CBE) formation from canabidiol (CBD), CBEM was formed from CBDM by the monooxygenase system including cytochrome P450. When cannabidiol dimethyl ether (CBDD), in which phenolic hydroxyl groups of CBD are masked with methyl groups, was incubated with liver microsomes and an reduced nicotinamide adenine dinucleotide phosphate-generating system, 1S,2R-epoxy-CBDD was identified by GC-MS. The epoxy metabolite was also found in the liver of a guinea pig pretreated with CBDD (100 mg/kg, intraperitoneally) 1 h before sacrifice. Rate of 1S,2R-epoxide metabolism was slower than that of 1R,2S-epoxy-CBDD under the conditions, as in the microsomal oxidation of CBDD described above. These results indicate that 1S,2R-epoxides are formed from CBD, CBDM and CBDD and that the epoxides are quickly converted to elsoin-type metabolites in the cases of CBD and CBDM.

Topics: Animals; Biotransformation; Cannabidiol; Guinea Pigs; In Vitro Techniques; Male

Biological formation of cannabielsoin (CBE) from cannabidiol (CBD) was studied in the guinea pig, mouse, rat and rabbit in vitro. Emphasis was placed on the elucidation of this formation mechanism. The enzyme activity of CBE formation was localized in hepatic microsomes. The enzymatic reaction required nicotinamide adenine dinucleotide phosphate (NADPH) and molecular oxygen, and showed an optimal pH around 7.3. The microsomal CBE-forming activities decreased in the following order; guinea pig greater than mouse greater than or equal to rabbit greater than or equal to rat. The CBE formation in the guinea pig hepatic microsomes was suppressed with various inhibitors of cytochrome P-450 such as SKF 525-A, alpha-naphthoflavone and carbon monoxide, but not by disodium ethylenediamine tetraacetate. When incubated with the microsomes either in the presence or absence of NADPH, a synthetic epoxide of CBD, 1S, 2R-epoxy-CBD-2',6'-diacetate was easily and exclusively converted to CBE. On the other hand, 1R, 2S-epoxy-CBD was not changed to CBE at all, but to several oxidized metabolites. These results suggest that CBD is biotransformed to 1S,2R-epoxy-CBD with hepatic microsomal monooxygenase system including cytochrome P-450, and the epoxide is immediately converted to CBE.

Topics: Animals; Benzoflavones; Cannabidiol; Cannabinoids; Chromatography, Gas; Cobalt; Epoxy Compounds; Gas Chromatography-Mass Spectrometry; Guinea Pigs; In Vitro Techniques; Male; Metyrapone; Mice; Mice, Inbred Strains; Microsomes, Liver; Pyridines; Rabbits; Rats; Rats, Inbred Strains; Species Specificity

Metabolism of cannabidiol (CBD), one of the major components of marihuana, was studied in the guinea pig both in vitro and in vivo. Analyses of metabolites by gas chromatography and gas chromatography-mass spectrometry proved that cannabielsoin (CBE) was formed from CBD as a novel metabolite, and that the amount was about one-sixth of 7-hydroxy-CBD, which was the most abundant metabolite under in vitro conditions in the presence of microsomal monooxygenase (cytochrome P-450). CBE was also found in the liver of the guinea pig that was given CBD (100 mg/kg) intraperitoneally 1 h before sacrifice. The effects of CBE on pentobarbital-induced sleep and body temperature were assessed in the mouse; CBE possessed little activity in either case.

Topics: Animals; Body Temperature; Cannabidiol; Cannabinoids; Chromatography, Gas; Gas Chromatography-Mass Spectrometry; Guinea Pigs; In Vitro Techniques; Male; Mice; Microsomes, Liver; Pentobarbital; Sleep