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S01 Subjective and Physiological Effects Following Controlled Smoked, Vaporized, and Oral Cannabis Administrations to Frequent and Occasional Cannabis Smokers

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S01 Subjective and Physiological Effects Following Controlled Smoked, Vaporized, and Oral Cannabis Administrations to Frequent and Occasional Cannabis Smokers S01 Subjective and Physiological Effects following Controlled Smoked, Vaporized, and Oral Cannabis Administrations to Frequent and Occasional Cannabis Smokers Matthew N. Newmeyer*1,2, Madeleine J. Swortwood1, Osama A. Abulseoud1, Marilyn A. Huestis1,3; 1Chemistry and Drug Metabolism, IRP, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD, 2Program in Toxicology, University of Maryland Baltimore, Baltimore, MD, 3University of Maryland School of Medicine, Baltimore, MD Background/Introduction: Cannabis is the most widely abused illicit drug worldwide. The active constituent Δ9- tetrahydrocannabinol (THC) may produce desirable euphoric effects, but negative subjective and physiological changes may also occur. While cannabis is most commonly administered via smoking, vaporization and oral consumption are becoming increasingly popular. However, subjective and physiological effects following these alternative administrations compared to smoking are not thoroughly characterized. Objective: Evaluate differences in subjective and physiological effects between smoked, vaporized, and oral cannabis in frequent and occasional cannabis smokers Method: Eleven frequent (≥5x/week) and nine occasional (≥2x but <3x/week) cannabis smokers provided informed written consent for this National Institute on Drug Abuse Institutional Review Board-, FDA-, and DEA-approved study. The study was randomized, double-blind, and placebo-controlled with a crossover and double-dummy design. Over 4 dosing sessions, participants were administered one active (6.9% THC; 54mg) or placebo cannabis-containing brownie followed by either one active or placebo cigarette or one active or placebo vaporized cannabis dose. No more than one active dose was administered per session. Visual analog scales consisting of 100-mm lines anchored by “not at all” and “most ever” were presented before and up to 5h post-dose for 9 items or up to 72h (frequent smokers) or 48h (occasional smokers) for 6 items; participants’ marked positions on the scales were converted to a percentage between 0-100. Heart rate (HR), systolic and diastolic blood pressure (SBP/DBP), and respiration rate (RR) were measured before and up to 3h post-dose. Data were analyzed via repeated-measures analysis of variance (ANOVA). Analyses were rerun with group data separated when significant group effects or interactions were present. Post-hoc tests with Bonferroni correction evaluated differences between administrations at each time point when significant interactions were observed. Finally, relationships between observed effects and matched blood THC concentrations were evaluated via linear mixed model analysis. Two-tailed p<0.05 indicated significance. Result: Ratings (times differences were observed) for “Good Drug Effect” (0.25-3.5h), “High” (0.5-2.5h), “Stoned” (0.25-2.5h) and “Stimulated” (0.25-1.5h) following smoking and vaporization were significantly higher compared to placebo for all participants. Only frequent smokers’ ratings for “Craving Marijuana” (0.25-3.5h) were significantly different from placebo following smoking and vaporization. Following oral cannabis administration, occasional smokers’ ratings for “Good Drug Effect” (1.5-3.5h), “High” (0.5-3.5h), and “Stoned” (overall dose effect observed) were significantly higher and ratings for “Willing to Drive-Nonemergency” (1.5-3.5h), and “Willing to Drive- Emergency” (1.5-3.5h) were significantly lower than after placebo. For frequent smokers only, ratings for “Willing to Drive-Emergency” were significantly lower (overall dose effect) following oral dosing compared to placebo. Participants’ HR changes from baseline at 0.5h following smoking (mean +11bpm) and vaporization (+10bpm) and changes from baseline at 1.5 (+9bpm) and 3.5h (+7bpm) following oral dosing were significantly larger than changes observed after placebo. Finally, significant relationships between blood THC concentrations and “Good Drug Effect”, “High”, “Stoned”, “Stimulated”, “Craving Marijuana”, and HR following smoking and vaporization were observed. Conclusion/Discussion: For the first time, differences in subjective and physiological effects following smoked, vaporized, and oral cannabis administrations were compared. Significant differences between smoked and vaporized cannabis were not present. Additionally, more group differences were observed following oral cannabis. These data help improve understanding of cannabis’ effects following alternative administrations. Keywords: Cannabis, Pharmacodynamics, Blood S02 Distribution of QT-Prolonging Drugs and Metabolites Between Cardiac Tissue and Femoral Blood in a Psychiatric Population Christian Reuss Mikkelsen1*, Jakob Ross Jornil1, Ljubica Vukelic Andersen1, Jytte Banner2, Jørgen Bo Hasselstrøm1; 1Department of Forensic Medicine, Aarhus University, Denmark, 2Department of Forensic Medicine, Copenhagen University, Denmark Background/Introduction: The Danish forensic autopsy-based prospective study, named SURVIVE, aims at elucidating the causes of the increased mortality and morbidity among mentally ill patients. Mentally ill patients are a heavily medicated patient group with high risk of early and sudden death. Many antipsychotics, antidepressants, and some substances of abuse consumed by mentally ill patients have been found to prolong the QT interval of the ECG. This can lead to the ventricular arrhythmia condition Torsades de Pointes, which in turn can lead to sudden cardiac death. In forensic postmortem toxicology, the blood concentrations are usually used for estimating a possible death by cardiac arrhythmia; however, animal studies and postmortem case studies have shown that some QT-prolonging drugs (QTDs) can be found in higher concentration in cardiac tissue when compared to peripheral blood. Objective: The aim of this project is to characterize the distribution of seven frequently used QTDs and their metabolites in cardiac tissue and femoral blood, and to investigate if the cardiac tissue concentration could be a tool for estimating cardiac arrhythmia as a potential cause of death. Method: Inclusion for SURVIVE was known or suspected mental illness and/or use of psychoactive medication at time of autopsy. Among the 500 cases included in SURVIVE, cases with at least one of QTD reported was selected for this project. A sample preparation method was developed and validated for the purpose of this project. After cardiac tissue homogenization, the cardiac homogenate and blood followed the same protein precipitation and filtration method. Deuterated internal standards were used. The extracts were analyzed on a Waters ACQUITY UPLC with a Waters Xevo TQ-S. Sex, age, BMI, manner and cause of death and time of last seen alive, declared death and autopsy as well as toxicology results were obtained from police, toxicology and autopsy reports for the descriptive analysis. Based on the toxicological evaluation, each compound result was classified A, B and C according to intoxication status. Result: Of the 500 SURVIVE cases, 211 cases were included in this project; 121 males and 90 females ranging from 18 to 94 years old. Each compound was found in between 8 and 90 cases each. The compound concentrations in whole blood and cardiac tissue were found to be statistically correlated (p<0.05) with 12 compounds very strong correlated (R^2>0.8), one strong correlated (R^2=0.76) and one moderate correlated (R^2= 0.62). Across all 14 compounds the heart-to-blood ratio (HB-ratio) ranged from 0.093 to 43 and the median HB-ratio ranged from 1.6 to 14. Each compound showed a high variance in HB-ratio between cases which was addressed statistically. Only age showed to be statistically significant in explaining the high HB-ratio variance of each compound. No difference in HB-ratios was detected due to extended postmortem interval. Conclusion/Discussion: In this project, we present the distribution of frequent used QTDs in the SURVIVE population between cardiac tissue and femoral blood. Keywords: QT-Prolonging Drugs, Heart-to-Blood Concentration Ratio, Forensic Toxicology S03 ELISA with Reflex to LC-MS/MS for Detection and Quantitation of Buprenorphine and Norbuprenorphine in Serum or Plasma Brandy M. Hill*1, Stephanie J. Marin2, Gwendolyn A. McMillin1,3; 1ARUP Laboratories, 2ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, 3University of Utah Department of Pathology, Salt Lake City, UT Background/Introduction: Buprenorphine is a partial opioid agonist-antagonist used to treat chronic pain and opioid addiction. Routine monitoring of buprenorphine compliance is recommended. Testing buprenorphine in blood avoids concerns about specimen adulteration that are common in high risk populations tested with urine. Here an enzyme- linked immunosorbent assay (ELISA) screen and confirmation assay performed by LC-MS/MS were validated to support clinical buprenorphine testing with serum/plasma. Steady state plasma concentrations of buprenorphine and norbuprenorphine in patients on a maintenance dose of buprenorphine (4-8 mg/day) can be as low as 0.8 ng/mL for buprenorphine and 1.1 ng/mL for norbuprenorphine (1). Glucuronide metabolites of buprenorphine and norbuprenorphine are present in serum and plasma. To maximize detection of buprenorphine and norbuprenorphine, enzyme hydrolysis and a cutoff of 1 ng/mL were implemented for the LC-MS/MS assay. Objective: To evaluate compliance for buprenorphine with the most sensitive assay possible. Method: An ELISA kit (part #236-0096, Immunalysis, Pomona, CA) was validated
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