MSACL 2018 US Abstract


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Topic: Cannabinoids

Method Development and Validation of LC-MS/MS Assay for Quantification of Cannabinoids in Whole Blood

Breland Smith (Presenter)
InSource Diagnostics

Bio: I completed my BS and MS in chemistry at Illinois State University (Normal, IL) specializing in organic chemistry with research in the area of novel porphyrin synthesis. I then received my Ph.D. in medicinal chemistry at the University of Arizona (Tucson, AZ) with research focused on the design, synthesis, and evaluation of small molecule drugs against novel translational targets for diseases including Alzheimers neurodegeneration and cancer. In July of 2017, I completed a two year ComACC accredited clinical chemistry post-doctoral fellowship at UCSD. I am an ASCP certified technologist in chemistry (ASCP, C) with a California CLS license. In September, I began working for InSource Diagnostics as general supervisor of Mass Spectrometry/Chemistry.

Authorship: Breland Smith, PhD
InSource Diagnostics

Short Abstract

A majority of US states have passed laws allowing either medical or recreational use of marijuana. The method development and validation of an LC-MS/MS assay to quantify ∆9-tetrahydrocannabinol (THC) and 9 other cannabinoids in whole blood will be discussed in this presentation. Cannabinoids are isolated from 200 µL of whole blood to achieve an LLOQ of 0.5 – 1.0 ng/mL for all analytes. This assay is currently being used to support studies in conjunction with the Center for Medical Marijuana Research (CMCR) aimed at understanding the pharmacokinetics of cannabinoids for the purpose of identifying biomarkers that can accurately predict impairment.

Long Abstract

Introduction

Currently, a majority of US states have passed some form of marijuana law, legalizing either recreational or medical use. Traditionally, marijuana use has been detected by measuring the presence of the 11-carboxy THC metabolite in urine which may stay elevated for several days to weeks after last administration, especially in chronic users. With marijuana legalization comes the responsibility to develop non-zero tolerance assays that accurately distinguish recent use and impairment from past use of marijuana leading to the presence of residual levels of metabolites that do not correlate with impairment.

Methods

The method development and validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) assay to quantify ∆9-tetrahydrocannabinol (THC), considered the main psychoactive component of marijuana, and 9 other cannabinoids in whole blood, specifically cannabinol (CBN), cannabidiol (CBD), 11-hydroxy-THC, (±)-11-nor-9-carboxy-Δ9-THC (THC-COOH), (+)-11-nor-Δ9-THC-9-carboxylic acid glucuronide (THC-COOH-gluc), cannabigerol (CBG), tetrahydrocannabivarin (THC-V), and tetrahydrocannabinolic acid (THCA-A) will be discussed in this presentation. 200 µL of whole blood is spiked with internal standard solution then cannabinoids are isolated through protein precipitation followed by solid phase extraction. After sample clean-up, evaporation, and reconstitution, 10 µL is injected onto a C18 UPLC column, and separated over a 3.5 minute gradient.

Results

A lower limit of quantitation (LLOQ) of 0.5 – 1.0 ng/mL was achieved for all analytes. A discussion of the studies to optimize assay sensitivity as well as the final validation data defining accuracy, precision, linearity, interferences, and matrix effects will be included in this presentation.

Conclusions & Discussion

This assay is currently being used to support studies in conjunction with the Center for Medical Marijuana Research (CMCR) aimed at understanding the pharmacokinetics of cannabinoids for the purpose of identifying biomarkers that can accurately predict impairment.


References & Acknowledgements:


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