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, 9 (4), 626-633

Simultaneous Quantification of Major Cannabinoids and Metabolites in Human Urine and Plasma by HPLC-MS/MS and Enzyme-Alkaline Hydrolysis

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Simultaneous Quantification of Major Cannabinoids and Metabolites in Human Urine and Plasma by HPLC-MS/MS and Enzyme-Alkaline Hydrolysis

Oier Aizpurua-Olaizola et al. Drug Test Anal.

Abstract

A high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) method for simultaneous quantification of Δ9-tetrahydrocannabinol (THC), its two metabolites 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH), and four additional cannabinoids (cannabidiol (CBD), cannabigerol (CBG), tetrahydrocannabivarin (THCV), and cannabinol (CBN)) in 1 mL of human urine and plasma was developed and validated. The hydrolysis process was studied to ensure complete hydrolysis of glucuronide conjugates and the extraction of a total amount of analytes. Initially, urine and plasma blank samples were spiked with THC-COOH-glucuronide and THC-glucuronide, and four different pretreatment methods were compared: hydrolysis-free method, enzymatic hydrolysis with Escherichia Coli β-glucuronidase, alkaline hydrolysis with 10 M NaOH, and enzyme-alkaline tandem hydrolysis. The last approach assured the maximum efficiencies (close to 100%) for both urine and plasma matrices. Regarding the figures of merit, the limits of detection were below 1 ng/mL for all analytes, the accuracy ranged from 84% to 115%, and both within-day and between-day precision were lower than 12%. Finally, the method was successfully applied to real urine and plasma samples from cannabis users. Copyright © 2016 John Wiley & Sons, Ltd.

Keywords: cannabinoids and metabolites; enzymatic-alkaline glucuronide hydrolysis; high performance liquid chromatography-mass spectrometry; plasma and urine analysis; solid-phase extraction.

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