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 journalArticle

2 Citations (Scopus)

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

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Cannabidiol
Cannabinoids
Tandem Mass Spectrometry
Liquid Chromatography
Epilepsy
Cannabinol
Medical Marijuana
Ammonium Hydroxide
Acids
Dronabinol
Glucuronides
Cannabis
Quality Control
Glass
Methanol
Seizures
Guidelines
Pharmaceutical Preparations
Proteins

Keywords

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

Cite this

@article{73c9543a3e02491384c4087299b3c8ed,
title = "Simultaneous Quantification of 13 Cannabinoids and Metabolites in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry in Adult Epilepsy Patients",
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.",
keywords = "CBD, HPLC-MS/MS, assay, cannabinoids, tandem mass spectrometry",
author = "Roslawski, {Michaela J.} and Remmel, {Rory P} and Ashwin Karanam and Leppik, {Ilo E} and Marino, {Susan E} and Birnbaum, {Angela K}",
year = "2019",
month = "6",
day = "1",
doi = "10.1097/FTD.0000000000000583",
language = "English (US)",
volume = "41",
pages = "357--370",
journal = "Therapeutic Drug Monitoring",
issn = "0163-4356",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Simultaneous Quantification of 13 Cannabinoids and Metabolites in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry in Adult Epilepsy Patients

AU - Roslawski, Michaela J.

AU - Remmel, Rory P

AU - Karanam, Ashwin

AU - Leppik, Ilo E

AU - Marino, Susan E

AU - Birnbaum, Angela K

PY - 2019/6/1

Y1 - 2019/6/1

N2 - 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.

AB - 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.

KW - CBD

KW - HPLC-MS/MS

KW - assay

KW - cannabinoids

KW - tandem mass spectrometry

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U2 - 10.1097/FTD.0000000000000583

DO - 10.1097/FTD.0000000000000583

M3 - Article

C2 - 30520828

AN - SCOPUS:85066060824

VL - 41

SP - 357

EP - 370

JO - Therapeutic Drug Monitoring

JF - Therapeutic Drug Monitoring

SN - 0163-4356

IS - 3

ER -