Research Paper

Analytical and Bioanalytical Chemistry

, Volume 405, Issue 14, pp 4679-4689

Impact of enzymatic and alkaline hydrolysis on CBD concentration in urine

  • Mateus M. BergamaschiAffiliated withChemistry and Drug Metabolism, IRP, National Institute on Drug Abuse, NIHSchool of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo
  • , Allan BarnesAffiliated withChemistry and Drug Metabolism, IRP, National Institute on Drug Abuse, NIH
  • , Regina H. C. QueirozAffiliated withSchool of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo
  • , Yasmin L. HurdAffiliated withPharmacology and Systems Therapeutics, Mount Sinai School of Medicine
  • , Marilyn A. HuestisAffiliated withChemistry and Drug Metabolism, IRP, National Institute on Drug Abuse, NIHChemistry and Drug Metabolism, Intramural Research Program, NIDA, NIH, Biomedical Research Center Email author 

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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.

Keywords

Cannabidiol Cannabidiol-glucuronide Enzymatic hydrolysis Urine GC-MS