Detecting and quantifying marijuana metabolites in serum and urine of 19 dogs affected by marijuana toxicity

Veterinarians diagnose marijuana toxicity based on clinical signs and history, or in conjunction with an over-the-counter (OTC) human urine drug screen. With the legalization of recreational marijuana use becoming more prevalent in the United States, a more accurate test to aid in the diagnosis of canine marijuana toxicity is needed. We collected urine and serum samples from 19 dogs with confirmed or suspected marijuana toxicosis from multiple veterinary hospitals and analyzed them with a novel UPLC-MS/MS method. Calibrations from 0.1 to 100 ng/mL and QC materials were prepared. Samples were extracted, purified, and eluted with solid-phase extraction. Urine samples were tested with an OTC human urine drug screen. The limit of detection (LOD) and lower limit of quantification (LLOQ) ranges for marijuana metabolites in serum were 0.05-0.25 ng/mL and 0.1-0.5 ng/mL, respectively. In urine, the LOD and LLOQ ranges for the metabolites were 0.05-0.1 ng/mL and 0.1-0.5 ng/mL, respectively. In serum, median and range of metabolite concentrations (ng/mL) detected included: THC, 65.0 (0.14-160); 11-OH-Δ⁹-THC, 4.78 (1.15-17.8); 11-nor-9-carboxy-Δ⁹-THC, 2.18 (0.71-7.79); CBD, 0.28 (0.11-82.5); and THC-glucuronide, 2.05 (0.72-18.3). In the 19 urine samples, metabolite: creatinine (ng: mg) values detected included: THC, 0.22 (0.05-0.74); 11-OH-Δ⁹-THC, 0; 11-nor-9-carboxy-Δ⁹-THC, 1.32 (0.16-11.2); CBD, 0.19 (0.12-0.26); THC-COOH-glucuronide, 0.08 (0.04-0.11); and THC-glucuronide, 0.98 (0.25-10.7). Twenty of 21 urine samples tested negative for THC on the urine drug screen. All 19 serum samples contained quantifiable concentrations of THC using our novel UPLC-MS/MS method. Utilizing a UPLC-MS/MS method can be a useful aid in the diagnosis of marijuana toxicosis in dogs, whereas using an OTC human urine drug test is not a useful test for confirming marijuana exposure in dogs because of the low concentration of THC-COOH in urine.