Simultaneous Quantification of 13 Cannabinoids and Metabolites in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry in Adult Epilepsy Patients

Michaela J. Roslawski, Rory P Remmel, Ashwin Karanam, Ilo E Leppik, Susan E Marino, Angela K Birnbaum

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Background:A sensitive, robust method was developed and validated to quantitate 13 major natural cannabinoid parent and metabolite compounds in human plasma at or below 0.5 ng/mL.Methods:A liquid chromatography tandem mass spectrometry method was developed and validated to measure 13 cannabinoid compounds: cannabidiol (CBD), cannabidiolic acid, cannabidivarin, cannabinol, cannabigerol, cannabigerolic acid, cannabichromene, Δ9-tetrahydocannabinol (THC), Δ9-tetrahydrocannabinolic acid A (THCA), Δ9-tetrahydrocannabivarin (THCV), 11-hydroxy-Δ9-tetrahydrocannbinol (11-OH-THC), 11-nor-9-carboxy-Δ9-tetrahydrocannbinol (THC-COOH), and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol glucuronide (THC-COOH-glu). Samples (200 L) were extracted through protein precipitation and separated with a Kinetex EVO C18 column and a 65%-95% gradient of methanol and 0.2% ammonium hydroxide/H2O at a flow rate of 0.4 mL/min. Samples were obtained from patients with epilepsy receiving cannabis for the treatment of seizures.Results:The extracted lower limit of quantification was 0.05 ng/mL for CBD, cannabidivarin, cannabinol, and 11-OH-THC; 0.10 ng/mL for cannabidiolic acid, cannabigerol, cannabichromene, cannabigerolic acid, THC, THCA, and THCV; and 0.50 ng/mL for THC-COOH and THC-COOH-glu. Mean quality control intraday accuracy and precision for all analytes ranged 96.5%-104% and 2.7%-4.9%, respectively, whereas interday accuracy and precision ranged 98%-103.3% and 0.2%-3.6%, respectively. An absolute matrix effect was observed for some analytes, however, with minimal relative matrix effect. Lack of nonspecific drug binding to extraction glass and plasticware was verified. Patient CBD levels ranged from 0.135 to 11.13 ng/mL.Conclusions:The validated method met FDA guidelines for bioanalytical assays precision and accuracy criteria. The assay reliably confirmed the use of particular medical cannabis formulations in patient samples as well as reliably measured low CBD concentrations from single-dose CBD exposure.

Original languageEnglish (US)
Pages (from-to)357-370
Number of pages14
JournalTherapeutic drug monitoring
Volume41
Issue number3
DOIs
StatePublished - Jun 1 2019

Bibliographical note

Funding Information:
A. K. Birnbaum reports grants from the Epilepsy Foundation, National Institutes of Health, Superunus Pharmaceuticals, and Veloxis Pharma-ceutics during the conduct of the study. She also has a royalty agreement for intravenous carbamazepine with Lundbeck. S. E. Marino reports grants from the Epilepsy Foundation, National Institutes of Health, Superunus Pharmaceuticals, and Veloxis Pharmaceutics during the conduct of the study. She and R. P. Remmel have a royalty agreement for intravenous topiramate Ligand Pharmaceuticals. The remaining authors declare no conflict of interest.

Funding Information:
Supported by the Patricia L Nangle Fund through the Epilepsy Foundation, and MacMillan Innovative Epilepsy Research and Education Fund.

Publisher Copyright:
© 2019 Wolters Kluwer Health, Inc.

Keywords

  • CBD
  • HPLC-MS/MS
  • assay
  • cannabinoids
  • tandem mass spectrometry

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