2 S-ketamine oral thin film pharmacokinetics S-ketamine and its metabolites, S-norketamine and S-hydroxynorketamine, were measured at the Department of Pharmacy and Toxicology using liquid chromatography coupled to QTOF-MS (hybrid quadrupole time-of-flight mass spectrometry) as detection technique (i.e. LC-QTOF- MS/MS). The LCQTOF-MS/MS system consisted of a Thermo Scientific double pump 3000 gradient system gradient with Bruker IL-2 QTOF.A column (Xterra MS C18 3.5µm x 2.1 mm x 100 mm) and precolumn (Xterra MS C18 Vanguard cartridge 3.5 µm x 2.1 mm) and was purchased from Waters Chromatography Europe BV (the Netherlands). For separation the mobile phase was methanol/water with 0.1 % formic acid with a gradient from 6.8-96 % methanol from 1 until 8.5 min. The total separation time was 15 min with a flow rate of 0.3 ml/min. The eluent was directed to the QTOF-MS from 1.2 until 7 min while the other part was directed to waste by a valve to avoid contamination of the QTOF. The system was controlled by Chromeleon Chromatography Data System software (Thermo Fisher Scientific, the Netherlands) for the LC part and Hystar (Bruker Nederland BV, the Netherlands) for the QTOF/MS part. In the positive ionization mode, the masses of the M+H ions were respectively 224.084, 228.109, 238.0993, 242, 124, 240.0786 and 246.099 Da for norketamine, norketamine-D4, ketamine, ketamine-D4, Cis-6-hydroxynorketamine and hydroxynorketamine13C6. Quant Analysis (Bruker Nederland BV, the Netherlands) was used for quantification of all analytes with a weighed (1/X*X) calibration line. The lower limits of quantitation were 6 ng/ml (0.025 nmol/mL), 6 ng/ml (0.026 nmol/mL) and 4 ng/ml (0.01 nmol/mL), for S-ketamine, S-norketamine and S-hydroxynorketamine, respectively. The upper limits of quantitation were, respectively, 500 ng/ml (2.1 nmol/ml), 1,000 (4.4 nmol/ml) and 200 ng/mL (0.72 nmol/ml) for S-ketamine, S-norketamine and S-hydroxynorketamine. Adverse Events Reported adverse events related to treatment were collected and were split up into events related to the 50 or 100 mg oral thin film or to the intravenous administration of S-ketamine. Additionally, the subjects were queried for dissociative side effects using the Bowdle questionnaire.9 TheBowdle questionnaire allows derivation of three factors of psychedelic ketamine effects: drug high and changes in internal and external perception. All three were measured on a visual analog score from 0 (no effect) to 10 cm (maximum effect). The questionnaire was first published in 1998 as a hallucinogen rating scale to quantify ketamine-induced psychedelic symptoms in volunteers and has been used in multiple studies on the effect of various psychedelics on dissociative symptoms. Blood pressure was obtained from the arterial-line using the FloTrac and Hemosphere system (Edwards Lifesciences, Irvine USA). 17
RkJQdWJsaXNoZXIy MTk4NDMw