MSACL 2025 Abstract

Development of Novel LC-MS/MS Psychoactive Panel (including Xylazine and Medetomidine) for Investigation of Prevalence in New Haven, CT Region

Jillian Kodger (1), Joe El-Khoury (1) , Gina Cassella-Mclane (2)
(1) Yale University , (2) Yale New Haven Health

Jill Kodger, PhD (Presenter) Yale University >> POSTER (PDF)

Presenter Bio: In 2024, I earned my PhD in Clinical-Bioanalytical Chemistry from Cleveland State University. My dissertation research focused on Barth Syndrome. As part of these studies, I had the opportunity to learn various techniques, including untargeted metabolomics and stable isotope tracing. I am currently a first-year Clinical Chemistry Fellow at Yale-New Haven Health System, with an interest in clinical mass spectrometry. My current research is focused on the development and application of novel psychoactive assays using LC-MS/MS, as well as the implementation of DART in the clinical setting. After completing my fellowship, I plan to pursue a career as a Clinical Laboratory Director at an academic medical center.

Relevant Financial Disclosures (within past 24 months, reported on Mar 19, 2025) No relevant financial relationship(s) to disclose. Conflict mitigation NOT REQUIRED.

INTRODUCTION:
Novel psychoactive substances are synthetic compounds designed to mimic the effects of commonly misused illicit drugs. This study aims to develop a method for detecting and quantifying emerging veterinary anesthetics, kratom, synthetic opioids, synthetic cannabinoids, and synthetic benzodiazepines. The inclusion of veterinary drugs in this study is driven by the increasing prevalence of xylazine and medetomidine in patients with polydrug abuse. Kratom, used by approximately 0.7% of the U.S. population, is often consumed by individuals with opioid use disorder or polydrug abuse, and kratom-related deaths are frequently linked to polydrug use. The rise of synthetic opioids, particularly fentanyl, has contributed to what is now recognized as the "fourth wave" of the opioid epidemic, highlighting the urgent need for effective testing methods for these substances. Synthetic cannabinoids, often referred to as "bath salts," are synthetic stimulants chemically related to the khat plant and are commonly used in vapes. These compounds have been found in overdose victims and are increasingly involved in toxicological cases. There has been an uptick in novel psychoactive drug use, particularly veterinary drugs, in New Haven and surrounding areas, prompting our efforts to quantify this in our polydrug user population for surveillance purposes.

METHODS:
Urine (150uL) and internal standard (150uL at 200 ng/mL), were combined in micro centrifuge tube, vortexed, and centrifuged and poured into sample vials. 2uL of the resulting solution was injected onto an Acquity HSS T3 column (Waters; 1.8um, 2.1 x 50mm) with an Acquity UPLC H33 T3 VanGuard pre-column (Waters; 1.8um) coupled to a Xevo TQD(Waters). The mass spectrometer was operated in positive ion mode. Quantification was based on peak area ratios of xylazine (m/z 221.19>89.90) to xylazine D6 (m/z 227.18>89.94), 7-OH mitragynine (m/z 415.32>190.02), mitragynine (m/z 399.25 >174.14) , Gabapentin (m/z 172.03 >54.90) to gabapentin 13C3( m/z 175.17> 140.17), metonitazene (m/z 383.25 > 100.07) to metonitazene 13C6( m/z 175.17> 140.17), etonitazene (m/z 397.25>100.01) to etonitazene 13C6 (m/z 403.30 > 99.95) and medetomidine (m/z 201.13 > 94.94) to medetomidine 13C-D3 (205.13 > 98.93).

RESULTS:
A seven minute gradient of 50-100% mobile phase B was sufficient to separate all compounds using water +0.1% formic acid (FA) (mobile phase A) and methanol +0.1% FA (mobile phase B). The analytical measureable range (AMR) was within %CV <20% and had a recovery between 80-120%. AMR was 5 to 1000 ng/mL for all analytes except gabapentin, which was 20-1000 ng/mL. No significant carryover was observed up to a concentration of 200 ng/mL for all analytes. In a small sampling of 48 patients in our polydrug user population we found around 2.1% prevalence of medetomidine, 27.1 % prevalence of xylazine and 33.3% prevalence of gabapentin.

CONCLUSION:
In conclusion, novel psychoactive substances are detectable in our polydrug user population in the New Haven area, with xylazine being the most common, followed by medetomidine.