Abstract
A sensitive and specific analytical method for cannabidiol (CBD) in urine was needed to define urinary CBD pharmacokinetics after controlled CBD administration, and to confirm compliance with CBD medications including Sativex—a cannabis plant extract containing 1:1 ∆9-tetrahydrocannabinol (THC) and CBD. Non-psychoactive CBD has a wide range of therapeutic applications and may also influence psychotropic smoked cannabis effects. Few methods exist for the quantification of CBD excretion in urine, and no data are available for phase II metabolism of CBD to CBD-glucuronide or CBD-sulfate. We optimized the hydrolysis of CBD-glucuronide and/or -sulfate, and developed and validated a GC-MS method for urinary CBD quantification. Solid-phase extraction isolated and concentrated analytes prior to GC-MS. Method validation included overnight hydrolysis (16 h) at 37 °C with 2,500 units β-glucuronidase from Red Abalone. Calibration curves were fit by linear least squares regression with 1/x 2 weighting with linear ranges (r 2 > 0.990) of 2.5–100 ng/mL for non-hydrolyzed CBD and 2.5–500 ng/mL for enzyme-hydrolyzed CBD. Bias was 88.7–105.3 %, imprecision 1.4–6.4 % CV and extraction efficiency 82.5–92.7 % (no hydrolysis) and 34.3–47.0 % (enzyme hydrolysis). Enzyme-hydrolyzed urine specimens exhibited more than a 250-fold CBD concentration increase compared to alkaline and non-hydrolyzed specimens. This method can be applied for urinary CBD quantification and further pharmacokinetics characterization following controlled CBD administration.
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Acknowledgments
The authors acknowledge Jeff Hackett from United Chemical Technologies for his technical support and Georgia Yiannoulos for her assistance with the research subjects. Intramural Research Program, National Institute on Drug Abuse, NIH, funded this research as well as a grant DA027781 to YLH and UL1RR029887. MMB received grant from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil).
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Bergamaschi, M.M., Barnes, A., Queiroz, R.H.C. et al. Impact of enzymatic and alkaline hydrolysis on CBD concentration in urine. Anal Bioanal Chem 405, 4679–4689 (2013). https://doi.org/10.1007/s00216-013-6837-x
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DOI: https://doi.org/10.1007/s00216-013-6837-x