Msd6140 single quadrodpole

Blood samples were collected (~500 μl) via jugular needle insertion following anesthesia with an isoflurane/oxygen vapor mixture (isoflurane 5% induction). Plasma THC content was quantified using fast liquid chromatography/mass spectrometry (LC/MS) adapted from (Irimia, Polis, Stouffer, & Parsons, 2015 (link); Lacroix & Saussereau, 2012 (link); Nguyen et al., 2018 (link)). 50 μl of plasma were mixed with 50 μl of deuterated internal standard (100 ng/ml CBD-d3 and THC-d3; Cerilliant), and cannabinoids were extracted into 300 μL acetonitrile and 600 μl of chloroform and then dried. Samples were reconstituted in 100 μl of an acetonitrile/methanol/water (2:1:1). Separation was performed on an Agilent LC1100 using an Eclipse XDB-C18 column (3.5um, 2.1mm × 100mm) using gradient elution with water and methanol, both with 0.2 % formic acid (300 μl/min; 73–90%). Cannabinoids were quantified using an Agilent MSD6140 single quadrodpole using electrospray ionization and selected ion monitoring [CBD (m/z=315.2), CBD-d3 (m/z=318.2), THC (m/z=315.2) and THC-d3 (m/z=318.2)]. Calibration curves were conducted daily for each assay at a concentration range of 0–200 ng/mL and observed correlation coefficients were 0.999.

Blood samples were collected (~300–500 μl) via jugular needle insertion following anesthesia with an isoflurane/oxygen vapor mixture (isoflurane 5% induction) or via implanted catheter for groups prepared for i.v. sampling. Plasma THC content was quantified using fast liquid chromatography/mass spectrometry (LC/MS) adapted from (Irimia, Polis, Stouffer, & Parsons, 2015; Lacroix & Saussereau, 2012; Nguyen et al., 2018 (link)). 50 μl of plasma were mixed with 50 μl of deuterated internal standard (100 ng/ml CBD-d3 and THC-d3; Cerilliant), and cannabinoids were extracted into 300 μL acetonitrile and 600 μl of chloroform and then dried. Samples were reconstituted in 100 μl of an acetonitrile/methanol/water (2:1:1). Separation was performed on an Agilent LC1100 using an Eclipse XDB-C18 column (3.5um, 2.1mm × 100mm) using gradient elution with water and methanol, both with 0.2 % formic acid (300 μl/min; 73–90%). Cannabinoids were quantified using an Agilent MSD6140 single quadrodpole using electrospray ionization and selected ion monitoring [CBD (m/z=315.2), CBD-d3 (m/z=318.2), THC (m/z=315.2) and THC-d3 (m/z=318.2)]. Calibration curves were conducted daily for each assay at a concentration range of 0–200 ng/mL and observed correlation coefficients were 0.999.