Simultaneous Quantification of 17 Cannabinoids by LC-MS-MS in Human Plasma

Area of Science:

• Pharmacology
• Analytical Chemistry
• Biochemistry

Background:

• Increasing use of tetrahydrocannabinol (THC) and cannabidiol (CBD) necessitates precise analytical methods for clinical applications.
• Understanding the pharmacokinetics and metabolism of cannabinoids is crucial for establishing dose-effect relationships.
• Existing analytical methods may lack the sensitivity or specificity required for comprehensive cannabinoid profiling in biological matrices.

Purpose of the Study:

• To develop and validate a sensitive and specific assay for the simultaneous quantification of 17 cannabinoids and their metabolites in human plasma.
• To identify and quantify major human metabolites of CBD.
• To provide a robust analytical tool for large-scale clinical studies involving cannabinoid therapeutics.

Main Methods:

• Development and validation of an online extraction high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) method.
• Simultaneous quantification of 17 cannabinoids, including THC and CBD metabolites, in human plasma.
• Utilized a one-step protein precipitation for sample preparation followed by LC/LC-atmospheric pressure chemical ionization-MS-MS analysis.

Main Results:

• The validated LC-MS-MS assay demonstrated a lower limit of quantification ranging from 0.78 to 7.8 ng/mL and upper limits between 100 and 2,000 ng/mL.
• Analytical accuracy and imprecision were within acceptable ranges (90.4–111% and 3.1–17.4%, respectively).
• Identified (3R-trans)-cannabidiol-7-oic acid (7-CBD-COOH), 7-hydroxy-CBD, and CBD-glucuronide (CBD-gluc) as major human CBD metabolites in plasma.

Conclusions:

• A robust and validated LC-MS-MS assay enables simultaneous quantification of multiple cannabinoids and their metabolites in human plasma.
• The assay successfully quantified CBD-glucuronide and identified key CBD metabolites, advancing the understanding of CBD's metabolic fate.
• This method has been effectively applied to over 5,000 clinical samples, supporting ongoing cannabinoid research.