title: Development of a simple and sensitive HPLC-UV method for the simultaneous determination of cannabidiol and Δ(9)-tetrahydrocannabinol in rat plasma.
abstract: There has been increased interest in the medical use of cannabinoids in recent years, particularly in the predominant natural cannabinoids, cannabidiol (CBD) and Δ(9)-tetrahydrocannabinol (THC). The aim of the current study was to develop a sensitive and reliable method for the quantification of CBD and THC in rat plasma. A combination of protein precipitation using cold acetonitrile and liquid-liquid extraction using n-hexane was utilised to extract CBD and THC from rat plasma. Samples were then evaporated and reconstituted in acetonitrile and 30 μL was injected into an HPLC system. Separation was achieved using an ACE C18-PFP 150 mm × 4.6 mm, 3 μm column at 55 °C with isocratic elution using a mobile phase consisting of acetonitrile-water (62:38, v/v) at 1 mL/min for 20 min. Both cannabinoids, as well as the internal standard (4,4-dichlorodiphenyltrichloroethane, DDT) were detected at 220 nm. Our new method showed linearity in the range of 10-10,000 ng/mL and a lower limit of quantification (LLOQ) of 10 ng/mL for both cannabinoids, which is comparable to previously reported LC-MS/MS methods. Inter- and intra-day precision and accuracy were below 15% RSD and RE, respectively. To demonstrate the suitability of the method for in vivo studies in rats, the assay was applied to a preliminary pharmacokinetic study following IV bolus administration of 5 mg/kg CBD or THC. In conclusion, a simple, sensitive, and cost-efficient HPLC-UV method for the simultaneous determination of CBD and THC has been successfully developed, validated and applied to a pharmacokinetic study in rats.
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK; College of Pharmacy, University of Anbar, Anbar, Iraq.
authors.forename: Atheer
authors.initials: A
authors.lastname: Zgair
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK.
authors.forename: Jonathan C M
authors.initials: JC
authors.lastname: Wong
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK.
authors.forename: Akmal
authors.initials: A
authors.lastname: Sabri
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Fischer
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Barrett
authors.affiliation: Division of Clinical Neuroscience, University of Nottingham and Queen's Medical Centre, Nottingham, UK.
authors.forename: Cris S
authors.initials: CS
authors.lastname: Constantinescu
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK. Electronic address: Pavel.Gershkovich@nottingham.ac.uk.
authors.forename: Pavel
authors.initials: P
authors.lastname: Gershkovich
mesh_terms: D000818:Animals; D002185:Cannabidiol / Q000032:analysis / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D025924:Chromatography, Supercritical Fluid; D013759:Dronabinol / Q000032:analysis / Q000097:blood*; D004340:Drug Contamination; D055772:Green Chemistry Technology; D006863:Hydrogen-Ion Concentration; D010949:Plasma / Q000378:metabolism; D051381:Rats; D012293:Rifampin / Q000032:analysis; D012680:Sensitivity and Specificity; D012997:Solvents / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013696:Temperature
chemical_list: D012997:Solvents; D002185:Cannabidiol; D013759:Dronabinol; D012293:Rifampin
keywords: Cannabidiol; HPLC–UV; Pharmacokinetics; Rat plasma; Δ(9)-Tetrahydrocannabinol